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P·01 Products / Immune checkpoint

R12 — anti-CD112R/PVRIG for brightfield and multiplex IHC.

Clone R12 · Cat. no. DIA-R12 · Directed against the receptor CD112R/PVRIG (distinct from the ligand CD112/NECTIN2)

CD112R inhibits T cell mediated signals. R12 has been validated on large quantities of normal and tumor tissues. The unique clone R12 is ideally suited for multiplexed IHC studies of CD112R/PVRIG expression in human tissue specimens.

Scientific USP

523 colorectal cancers — peer-reviewed multiplex IHC

Yang 2023 · Modern Pathology · clone R12 / DIA-R12 named

  • Receptor CD112R/PVRIG — distinct from the ligand CD112
  • Brightfield + multiplex · human FFPE
  • High signal-to-noise ratio

The IHC detection of PVRIG in tumor tissues so far is only possible with clone R12.

ONCOdianova home page (antibody highlight) and CD112R product page · Datasheet 25 Feb 2025/04 · IHC gallery · Protocols · Publications — panel conditions as published (AR pH 7.8 · 1:500 · Opal 570).

Mouse monoclonal antibody for studying the inhibitory receptor CD112R (PVRIG) in human formalin-fixed, paraffin-embedded tissue. Documented for brightfield IHC on FFPE sections (IHC-P) and fluorescence multiplex IHC (multiplex immunofluorescence, mIF), with tonsil as positive control and a membranous lymphocyte staining pattern; clone R12 combines high specificity with an inherently high signal-to-noise ratio and is well suited for multiplex IHC studies on human tissues. Clone R12 (DIA-R12) was used in a peer-reviewed multiplex study of 523 colorectal cancers and in two doctoral theses of the University of Hamburg. Documented applications: IHC on FFPE tissue (datasheet), fluorescence multiplex IHC (datasheet reactivity text, ONCOdianova product documentation and gallery) and Western blot (ONCOdianova product documentation).

  • Host / IsotypeMouse · IgG1/κ
  • ReactivityHuman
  • ApplicationsIHC on FFPE tissue · multiplex fluorescence IHC · Western blot
  • IHC starting range1:100 – 1:200
  • Positive controlTonsil · membranous
  • Format100 µl, lyophilized
View product dataIHC galleryIHC protocolsOrderContact
Datasheet_DIA-R12_anti-CD112RMSDS
Product image of the ONCOdianova anti-CD112R/PVRIG antibody clone R12: fluorescence multiplex immunohistochemistry of a normal human tonsil with anti-CD8 (DIA-TC8) and anti-CD112R/PVRIG clone R12 (DIA-R12)
Product image · Clone R12 · DIA-R12 · CD8 × CD112R multiplex
Evidence layer 01

Clone-specific datasheet

Datasheet DIA-R12 (version 25 Feb 2025/04; product content identical to V03, 22 Feb 2022) documents identity, immunogen, formulation, reconstitution, storage, FFPE-IHC starting conditions, tonsil control and membranous pattern.

Evidence layer 03

Documented applications and images

Brightfield IHC (IHC-P) and fluorescence multiplex IHC (mIF) on human FFPE tissue and Western blot (ONCOdianova product documentation); 15 original ONCOdianova figures (brightfield, CD112R × CD8, CD112R × FOXP3), a manual protocol and the published multiplex conditions.

Brightfield and multiplex gallery: section P·04 ↓

P·02 Technical data

Clone R12 technical specifications.

Product data

Cat.No.DIA-R12
SpecificityCD112R
Species ReactivityHuman
Host SpeciesMouse
IsotypeIgG1/κ
CloneR12
ApplicationImmunohistochemistry (IHC), Formalin-fixed Paraffin-embedded sections (FFPE), Western Blot
Conjugationunconjugated
DilutionIHC 1:100-1:200
FormatLyophilisate, Antibody purified (from culture supernatant)
Quantity100 µl
Positive controlTonsil
PresentationPBS pH 7.4, 2% BSA, 0.05% NaN3 — BSA-free high conc. variant available upon request
Intended UseFor Research Use Only. Not for use in diagnostic procedures.
Manufacturer / BrandONCOdianova GmbH

Complete product data

ProductAnti-CD112R/PVRIG (Human) from Mouse
CloneR12 (monoclonal)
Catalog numberDIA-R12
Distributor catalog numberODN-DIA-R12 (BIOZOL)
Product categoryCancer Immunology IHC marker
TargetCD112R / PVRIG — poliovirus receptor-related immunoglobulin domain-containing protein, the CD112 receptor (Gene ID 79037, UniProt Q6DKI7). Not the ligand CD112/NECTIN2/PVRL2.
ImmunogenRecombinant peptide
IsotypeMouse IgG1/κ
Host speciesMouse
ReactivityHuman
ConjugationUnconjugated
Format100 µl, lyophilized; antibody purified from culture supernatant
Quantity100 µl
ReconstitutionRestore to 100 µl with sterile distilled water; gentle shaking for 10 minutes
FormulationPBS, pH 7.4, 2% BSA, 0.05% sodium azide
ApplicationsIHC on standard FFPE sections, IHC-P (datasheet) · fluorescence multiplex IHC / multiplex immunofluorescence, mIF (datasheet reactivity text; clone-specific publications; gallery) · Western blot (ONCOdianova product documentation; the Jansen thesis additionally reports a Western blot specificity check of DIA-R12)
IHC starting range1:100 – 1:200 for polymer-based detection (datasheet); 1:150 in the manual protocol of the ONCOdianova product documentation; working dilution established by each laboratory for its tissue, fixation, platform and detection system
Epitope retrievalHeat-induced epitope retrieval required; autoclave 121 °C, 5 min, Tris-EDTA-citrate pH 7.8 recommended
Primary antibody incubation60 min at 37 °C (datasheet and ONCOdianova product documentation)
Positive controlTonsil
Staining patternMembranous (lymphocyte subsets)
Automated platformsSuited for automated IHC platforms (datasheet)
StorageLyophilized at 2–8 °C; long-term at −20 °C (stable for at least one year); reconstituted at 2–8 °C short term (several weeks); avoid repeated freeze/thaw cycles
Associated antibodiesDIA-TC8 (anti-CD8, clone TC8) · DIA-TG1 (anti-TIGIT, clone TG1) — as listed in the datasheet
StatusFor Research Use Only. Not for use in diagnostic procedures.

The technical specifications above are documented in datasheet DIA-R12 (version 25 Feb 2025/04), whose product content is identical to V03 (22 Feb 2022), and on the ONCOdianova product documentation where indicated; gene and protein entries link to their registries. The ligand CD112/NECTIN2 is named to distinguish it from the target. Each laboratory establishes its own working conditions on this documented basis.

P·03 Reactivity

Reactivity of clone R12.

Reactivity

Anti-PVRIG/CD112R clone R12 has been developed for detection of CD112R in routine formalin-fixed paraffin-embedded tissue specimen (IHC FFPE) to be used in bright field immunohistochemistry and moreover for multicolor immunofluorescence (fluorescence multiplex IHC). Anti-PVRIG/CD112R clone R12 has been validated for sensitivity and specificity on large quantities of normal and tumor tissues. Clone R12 displays no background in non lymphoid cells and in epithelial cells. Based on the high specificity combined with an inherent high signal to noise ratio clone R12 is ideally suited for multiplexed immunohistochemistry studies of CD112R in human tissue specimen.

CD112R, a member of poliovirus receptor–like proteins, is preferentially expressed on T-cells and inhibits T-cell receptor mediated signals. Blockade of the CD112R-CD112 interaction enhances T cell response. CD112R binds to its ligand CD112 which is widely expressed on antigen-presenting cells and on tumor cells. CD112R competes with CD226 to bind to CD112 and thereby acts as a coinhibitory receptor for T cells. Blockage of the CD112R–CD112 interaction enhances human T cell response.

TIGIT and CD226 constitute a T cell cosignaling pathway in which CD226 serves as the costimulatory and TIGIT as the coinhibitory receptor for the ligands CD155 and CD112. This TIGIT signaling axis includes a complex receptor ligand system with the marker CD112R, which has become a promising target in cancer immuno-therapy.

Species reactivityHuman
Host speciesMouse
Structures stained by R12 in FFPE tissueCell membranes of lymphocyte subsets
Positive controlTonsil — normal human tonsil with numerous CD112R/PVRIG-positive lymphocytes
Staining patternMembranous
ApplicationsBrightfield IHC on human FFPE tissue; fluorescence multiplex IHC; Western blot

P·05 IHC protocols & published application examples

IHC protocols and published application examples for clone R12.

Manual brightfield IHC

Anti-CD112R/PVRIG clone R12 can be used for immunohistochemical detection of CD112R in formalin-fixed, paraffin-embedded human tissue. A manual staining protocol using autoclave-based antigen retrieval has been established for clone R12.

Antigen retrieval
pH 7.8 · 121 °C · 5 min
Primary antibody
Clone R12 · 1:150 · 60 min at 37 °C
Detection
EnVision HRP Rabbit/Mouse · 30 min at 37 °C
Recommended positive control
Human tonsil

Source: documented ONCOdianova application protocol for DIA-R12 (block B).

Published multiplex fluorescence IHC application

An academic application of clone R12 has been documented at the University Medical Center Hamburg-Eppendorf in a multiplex fluorescence immunohistochemistry workflow. The associated research investigated CD112R within the immune microenvironment of colorectal cancer using multiplex fluorescence immunohistochemistry.

Antibody
DIA-R12 · clone R12
Dilution
1:500
Antigen retrieval
pH 7.8
Fluorophore
Opal 570
Multiplex panel
CD112R · CD8 · PD-1 · Ki67

Sources: Yang C., dissertation, Universität Hamburg / UKE 2021 (Table 3) · Yang et al., Mod Pathol 2023 · doi:10.1016/j.modpat.2022.100089

The protocols and published workflows shown here are documented application examples from their respective sources; the documented conditions provide practical starting points for laboratory establishment of DIA-R12 workflows.

Three documented application contexts: the current datasheet (A), the manual protocol of the ONCOdianova product documentation (B) and the multiplex conditions reported in the clone-specific studies (C). Each dilution is presented with its corresponding workflow and detection system. These documented conditions provide practical starting points for laboratory establishment of R12 IHC workflows.

A · DatasheetImmunohistochemical staining of standard formalin-fixed paraffin sectionsDatasheet DIA-R12 · version 25 Feb 2025/04 (identical wording in V03, 22 Feb 2022)

Preparation and epitope retrieval

  1. Deparaffinize and rehydrate according to standard procedures
  2. Heat-induced epitope retrieval (HIER) is required
  3. Recommended: autoclave at 121 °C (5 min) in Tris-EDTA-citrate, pH 7.8 (e.g. TEC buffer)

Primary antibody and detection

  1. Incubate primary antibody for 60 min at 37 °C
  2. Biotin/(strept)avidin-based detection possible (e.g. Vectastain® Elite® ABC-HRP kit/AEC)
  3. Polymer-based detection (e.g. Dako EnVision™ detection system, peroxidase/DAB): antibody at 1:100–1:200
  4. Suited for automated staining platforms (datasheet)
  5. Run positive and negative controls in parallel; positive control: tonsil; expected pattern: membranous
B · ONCOdianova product documentationManual stain with autoclaveONCOdianova product documentation DIA-R12, section “IHC protocols”

Staining protocols for anti-human CD112R antibody clone R12 · Cat.No.: DIA-R12 · Isotype: Mouse IgG1/k · Specificity: Human CD112R / PVRIG · Physical State: Lyophilized powder

  1. Reconstitution: restore DIA-R12 to 100 µl with sterile distilled water, gentle shaking for 10 minutes
  2. Pretreatment buffer: 121 °C / 5 min / pH 7.8
  3. Incubation primary antibody: 60 min / 37 °C, dilution 1:150
  4. EnVision HRP rabbit/mouse: 30 min / 37 °C

The ONCOdianova product documentation adds: “Applicable for automated staining procedures and validated for multicolor immunofluorescence (multiplexed IHC)”. Suitability for automated procedures is also stated in the datasheet; the multiplex application is documented by the clone-specific studies (block C) and the gallery. The 1:150 dilution lies within the datasheet range of 1:100–1:200.

C · Published study conditionsFluorescence multiplex IHC with DIA-R12 — published application contextYang et al., Mod Pathol 2023, Table 1 · Yang, doctoral thesis UKE 2021, Table 3 · Jansen, doctoral thesis UKE 2020, section 2.3

Yang et al. 2023 (TMA cohort of 523 colorectal cancers; large sections of 20 cases)

  1. Freshly cut 4-µm consecutive sections; Opal Dye Kit (Akoya Biosciences, Cat. #NEL811001KT)
  2. CD112R: Dianova, clone R12, Cat# DIA-R12 · antigen retrieval pH 7.8 · dilution 1:500 · staining position 2 · Opal 570
  3. Other panel markers of the study: Ki67 (position 1), PD-1 (position 3), CD8 (position 4) – reagents of other manufacturers, see publication
  4. Image acquisition: Leica Aperio VERSA 8 epifluorescence scanner; deep learning–based cell segmentation

Jansen 2020 (122 tumors from seven entities, 17 normal tissues)

  1. 4-µm consecutive sections of 4-mm microenvironment TMAs and large sections; OPAL Dye Kit (Perkin Elmer, Cat. #OP7DS1001KT) following the Akoya protocol
  2. Epitope retrieval in an autoclave for 30 min at 100–120 °C in buffer pH 9 (protocol of the thesis); antibody stripping in the microwave (5 min 100 °C, 5 min 93 °C) between cycles; DAPI counterstain
  3. Anti-CD112R clone DIA-R12 (Dianova, Hamburg); specificity examined in the thesis by Western blot, ELISA and pre-absorption with CD112R protein; membranous staining pattern reported

The 1:500 dilution and the Opal panel position belong to the described multiplex protocol; the datasheet range of 1:100–1:200 applies to brightfield IHC with polymer detection. Each source is presented with its own workflow.

P·06 Interpretation guide

How to read the staining.

Expected pattern

Membranous, lymphocyte subsets

The datasheet describes a membranous pattern with tonsil as positive control. In the published studies with DIA-R12, CD112R signal was reported at variable intensity on CD8+, CD4+, FOXP3+ and CD56+ cells but not on CD11c+ dendritic cells or CD68+ macrophages (Jansen 2020/2021), and was elevated on CD8+ T cells within T-cell nests of colorectal cancer (Yang 2023). These findings from the respective study cohorts illustrate the lymphocyte subsets in which CD112R can be evaluated with clone R12.

Documented validation scope

High signal-to-noise CD112R detection in human tissue

Clone R12 was validated for sensitivity and specificity on a large number of normal and tumor tissues and showed no background staining in non-lymphoid and epithelial cells. It combines high specificity with an inherently high signal-to-noise ratio (ONCOdianova product information), well suited for multiplex IHC studies. The Jansen thesis reports that DIA-R12 was examined by Western blot, ELISA and pre-absorption of the primary antibody with CD112R protein before multiplex use, and describes a consistently membranous pattern; Yang 2023 used DIA-R12 in a peer-reviewed multiplex panel.

Research use

Research interpretation of CD112R staining

R12 visualises the inhibitory receptor CD112R/PVRIG on the membrane of lymphocyte subsets in human FFPE sections. Researchers can evaluate CD112R-positive cells by distribution and tissue compartment and, in multiplex panels, by co-expression with markers such as CD8, PD-1, Ki67 or FOXP3, according to the study design. R12 has been used in fluorescence multiplex IHC to study CD112R expression on CD8+ tumor-infiltrating lymphocytes (TILs) within the human tumor microenvironment (Jansen, 2020). The documented evidence for R12 consists of datasheet DIA-R12, the ONCOdianova product documentation and gallery, the peer-reviewed multiplex study naming DIA-R12 (Yang 2023), the doctoral theses of Yang (2021) and Jansen (2020) and two AACR 2020 congress abstracts of the same group.

P·07 Target background

CD112R/PVRIG in research.

CD112R, also called PVRIG (poliovirus receptor-related immunoglobulin domain-containing protein; gene PVRIG, formerly C7orf15), is a transmembrane receptor of the poliovirus receptor/nectin family. It was identified as a coinhibitory receptor on human T cells that binds CD112 (nectin-2) with high affinity and competes with the costimulatory receptor CD226 (DNAM-1) for this ligand; disrupting the CD112R–CD112 interaction enhanced human T-cell responses in vitro.[1]

CD112 is broadly expressed on antigen-presenting cells and on tumor cells. PVRIG and its ligand PVRL2/CD112 are induced in cancer and inhibit CD8+ T-cell function,[4] and blockade of CD112R together with TIGIT sensitizes human NK-cell functions.[3] Together with TIGIT, CD226, CD96 and their ligands CD155 (PVR) and CD112, CD112R forms a receptor–ligand network that is studied as an immune checkpoint axis in cancer.[2, 5]

These statements describe the target biology. Clone R12 is the ONCOdianova research tool for tissue-based CD112R/PVRIG studies within this receptor–ligand network: it visualises CD112R/PVRIG-positive lymphocyte subsets in human FFPE tissue, alone or together with markers such as CD8, PD-1, Ki67 and FOXP3 in multiplex panels.

  • GenePVRIG — NCBI Gene 79037
  • ProteinTransmembrane protein PVRIG / CD112 receptor — UniProt Q6DKI7
  • AliasesCD112R · PVRIG · C7orf15
  • LigandCD112 (nectin-2; gene NECTIN2, formerly PVRL2) — the ligand, a distinct molecule; R12 is directed against the receptor
  • Competing receptorCD226 (DNAM-1) — Zhu et al. 2016 [1]
  • Related axisTIGIT / CD226 / CD96 with CD155 (PVR) and CD112 — see the ONCOdianova anti-TIGIT clones TG1 and TG2

Bracketed numbers refer to the target-literature list below. Clone-specific evidence for R12 is listed under Publications.

P·08 Translational research relevance

PVRIG/CD112R in translational immuno-oncology research.

Target-level

Translational context

PVRIG (identical to CD112R) is an inhibitory checkpoint of the DNAM-1 axis; its ligand is PVRL2 (CD112, nectin-2). COM701 is an investigational therapeutic anti-PVRIG antibody studied in registered phase 1 and phase 1/2 trials, which makes tissue PVRIG expression a target of translational interest.

Clone

What the clone can be used to study

Clone R12 can be used to study CD112R/PVRIG-positive lymphocyte subsets in human FFPE tissue, alone or in multiplex panels; it is named in a peer-reviewed four-marker fluorescence multiplex study of 523 colorectal cancers.

Target-level

Translational research context

Target-level studies, the registered COM701 trials and the immunohistochemical biomarker analysis of trial biopsies provide translational context for tissue PVRIG research; PVRIG IHC on FFPE tissue is part of the biomarker work that accompanies clinical PVRIG blockade.

IHC biomarkers in immunotherapy research Related research context: TG1 — TIGIT · TG2 — TIGIT · KK3 — CD73 · TC8 — CD8 · FX3 — FOXP3

P·09 Publications

Documented evidence.

Clone-specific evidence (clone R12 / DIA-R12)

  1. Yang C, Mandelkow T, Bady E, Raedler JB, Simon R, Sauter G, Lennartz M, Büscheck F, Luebke AM, Dum D, Menz A, Höflmayer D, Weidemann S, Fraune C, Lebok P, Uhlig R, Bernreuther C, Jacobsen F, Clauditz TS, Wilczak W, Minner S, Burandt E, Steurer S, Blessin NC. Nonredundant Upregulation of CD112R (PVRIG) and PD-1 on Cytotoxic T Lymphocytes Located in T Cell Nests of Colorectal Cancer. Modern Pathology (2023); 36(4):100089. doi:10.1016/j.modpat.2022.100089 · PMID 36788088
    Fluorescence multiplex IHC (CD8, PD-1, CD112R, Ki67) on FFPE tissue: a tissue-microarray cohort of 523 colorectal cancers with clinical follow-up and large sections of 20 colorectal cancers; R12 at antigen retrieval pH 7.8, 1:500, staining position 2, Opal 570.    Peer-reviewed article · clone R12 / DIA-R12
  2. Yang C. Upregulation of CD112R (PVRIG) and PD-1 on cytotoxic T-cells located in T-cell niche of colorectal cancer. Doctoral thesis, University of Hamburg / Institute of Pathology, University Medical Center Hamburg-Eppendorf (2021 (oral defense 17 Dec 2021)); URN urn:nbn:de:gbv:18-ediss-97788. Repository (University of Hamburg)
    Fluorescence multiplex IHC on colorectal cancer tissue with DIA-R12 in the Opal panel; the doctoral thesis of the first author of entry 1 on the same topic and from the same research group.    Academic thesis · clone R12 / DIA-R12
  3. Jansen HL. Expressionsunterschiede des Immuncheckpoint-Rezeptors CD112R in humanen Karzinomen im Vergleich zu Normalgeweben. Doctoral thesis, University of Hamburg / Institute of Pathology, University Medical Center Hamburg-Eppendorf (2020 (oral defense 27 Jul 2021)); URN urn:nbn:de:gbv:18-ediss-94757. Repository (University of Hamburg)
    Fluorescence multiplex IHC on 4-mm microenvironment TMAs of 122 tumors (seven entities) and large sections of 17 normal tissues; CD112R examined on CD8+, CD4+, FOXP3+, CD56+, CD11c+ and CD68+ cells; specificity of DIA-R12 examined by Western blot, ELISA and pre-absorption with CD112R protein.    Academic thesis · DIA-R12

Specific references from the American Association of Cancer Research (AACR) annual meeting 2020

Proceedings: AACR Annual Meeting 2020; April 27-28, 2020 and June 22-24, 2020; Philadelphia, PA, Volume 80, Issue 16 Supplement, pp. 3870 and pp. 4970

  1. Blessin NC, Mandelkow T, Bady E, Hube-Magg C, Sauter G, Simon R, Fraune C, Lennartz M, Weidemann SA, Möller K, Höflmayer D. Abstract 3870: Patterns of CD112R expression in normal lymphatic tissues, inflammation and cancer. Cancer Research (2020); 80(16 Suppl): 3870. doi:10.1158/1538-7445.AM2020-3870Congress abstract · AACR 2020
  2. Simon R, Blessin NC, Mandelkow T, Bady E, Hube-Magg C, Sauter G, Lennartz M, Fraune C, Weidemann SA, Möller K, Büscheck F. Abstract 4970: Prognostic role of CD112R, PD-1 and Ki67 expression in CD8+cytotoxic T cells in colorectal cancer. Cancer Research (2020); 80(16 Suppl): 4970. doi:10.1158/1538-7445.AM2020-4970Congress abstract · AACR 2020

The published abstract text of both references describes CD112R multiplex IHC by the same Hamburg group that later published Yang et al. 2023.

Target literature cited in the datasheet

  1. Zhu Y et al. Identification of CD112R as a novel checkpoint for human T cells. Journal of Experimental Medicine (2016). doi:10.1084/jem.20150785 · PMID 26755705Target biology
  2. Stamm H, Wellbrock J, Fiedler W. Interaction of PVR/PVRL2 with TIGIT/DNAM-1 as a novel immune checkpoint axis and therapeutic target in cancer. Mammalian Genome (2018). doi:10.1007/s00335-018-9770-7 · PMID 30132062Target biology · review
  3. Xu F et al. Blockade of CD112R and TIGIT signaling sensitizes human natural killer cell functions. Cancer Immunology, Immunotherapy (2017). doi:10.1007/s00262-017-2031-x · PMID 28623459Target biology
  4. Whelan S et al. PVRIG and PVRL2 Are Induced in Cancer and Inhibit CD8+ T-cell Function. Cancer Immunology Research (2019). doi:10.1158/2326-6066.CIR-18-0442 · PMID 30659054Target biology
  5. Sanchez-Correa B et al. DNAM-1 and the TIGIT/PVRIG/TACTILE Axis: Novel Immune Checkpoints for Natural Killer Cell-Based Cancer Immunotherapy. Cancers (2019). doi:10.3390/cancers11060877 · PMID 31234588Target biology · review

Entry 1 is the peer-reviewed clone-specific publication; entries 2 and 3 are academic theses cited with their repository records. The AACR abstracts and the target literature provide scientific context.

P·10 FAQ

Frequent scientific questions.

What is clone R12, and what does DIA-R12 identify?

R12 is a mouse monoclonal antibody (IgG1/κ) raised against a recombinant peptide of human CD112R, also called PVRIG (gene PVRIG). DIA-R12 is the ONCOdianova catalogue number of this antibody; the distributor catalogue number at BIOZOL is ODN-DIA-R12. The antibody is supplied as 100 µl lyophilisate (PBS, 2 % BSA, sodium azide) and is documented for immunohistochemistry on human formalin-fixed, paraffin-embedded (FFPE) tissue in brightfield IHC and fluorescence multiplex IHC, with tonsil as positive control.

What is the difference between CD112, CD112R, PVRIG and NECTIN2?

PVRIG and CD112R are two names for the same molecule: an inhibitory immune-checkpoint receptor of the poliovirus receptor/nectin family (gene PVRIG, formerly C7orf15) that is expressed in T-cell and NK-cell biology. CD112 and NECTIN2 (formerly PVRL2) are likewise two names for one molecule: the ligand of this receptor, expressed on antigen-presenting cells and tumor cells. Receptor and ligand are distinct proteins encoded by different genes. CD112R/PVRIG binds CD112 and competes with the costimulatory receptor CD226 (DNAM-1) for this ligand (Zhu et al. 2016). Clone R12 is directed against the receptor CD112R/PVRIG.

Is R12 suitable for immunohistochemistry on human FFPE tissue (IHC-P)?

Yes. Immunohistochemistry on formalin-fixed, paraffin-embedded sections (IHC-P) is the core documented application of clone R12. The datasheet specifies heat-induced epitope retrieval, a starting dilution of 1:100 to 1:200 for polymer-based detection, tonsil as positive control and a membranous staining pattern of lymphocyte subsets; the ONCOdianova product documentation documents a manual protocol with 1:150 and EnVision detection. Seven brightfield figures in the gallery on this page show CD112R staining with clone R12 in human FFPE tissue.

Which publications document the use of clone R12?

One peer-reviewed journal article used clone R12 (catalogue number DIA-R12): Yang et al., Modern Pathology 2023 (PMID 36788088), a fluorescence multiplex IHC study of 523 colorectal cancers. Two doctoral theses of the University of Hamburg used DIA-R12: Yang (2021) on CD112R and PD-1 in colorectal cancer and Jansen (2020) on CD112R expression in carcinomas compared with normal tissues; both are academic theses cited with their repository records. Two AACR 2020 congress abstracts of the same Hamburg group, listed as specific references on the ONCOdianova product documentation, describe CD112R multiplex IHC and are cited as congress abstracts. All entries with links are listed under Publications.

Is R12 documented for multiplex immunofluorescence (mIF)?

Yes. Fluorescence multiplex IHC, also termed multiplex immunofluorescence (mIF), is the published clone-specific application of DIA-R12: Yang et al. 2023 used the antibody within an Opal panel (antigen retrieval pH 7.8, dilution 1:500, Opal 570) on FFPE colorectal cancer tissue, and the theses of Yang (2021) and Jansen (2020) describe Opal-based multiplex workflows with DIA-R12. The datasheet reactivity text describes clone R12 as developed for brightfield IHC and moreover for multicolor immunofluorescence, and the ONCOdianova gallery documents CD112R × CD8 and CD112R × FOXP3 multiplex staining of human tonsil. The published conditions belong to their study panels and provide documented starting points for panel establishment.

Can R12 be combined with CD8 or FOXP3 in a multiplex panel?

Yes, both combinations are documented. A published fluorescence multiplex panel combined CD112R (clone R12) with CD8, PD-1 and Ki67 on FFPE colorectal cancer tissue (Yang et al. 2023), and the Jansen thesis (2020) evaluated CD112R on CD8+, CD4+, FOXP3+, CD56+, CD11c+ and CD68+ cells. ONCOdianova figures document CD112R × CD8 multiplex staining with clones R12 and TC8 and CD112R × FOXP3 multicolor immunofluorescence with clone R12 of normal human tonsil. Antibody order, retrieval, dilution and detection are established per panel; the published conditions provide a documented starting point.

What dilution conditions are documented for R12?

Three documented values, each within its own workflow: the datasheet states 1:100 to 1:200 for polymer-based peroxidase/DAB detection on FFPE sections (brightfield IHC-P); the manual protocol of the ONCOdianova product documentation used 1:150 with EnVision HRP detection; the published multiplex study used 1:500 within an Opal panel with Opal 570 (Yang et al. 2023). Each laboratory establishes its working dilution for its tissue, fixation, platform and detection system on this documented basis.

What antigen-retrieval conditions are documented for R12?

Heat-induced epitope retrieval is required. The datasheet recommends an autoclave at 121 °C for 5 minutes in Tris-EDTA-citrate buffer, pH 7.8 (TEC buffer); the manual protocol of the ONCOdianova product documentation uses the same pretreatment (121 °C, 5 min, pH 7.8). In the published multiplex study, the R12 step used antigen retrieval at pH 7.8 (Yang et al. 2023); the Jansen thesis (2020) describes autoclave retrieval at pH 9 within its Opal workflow. Details are given in the protocol section.

Which positive-control tissue and staining pattern are documented for R12?

Tonsil is the positive-control tissue listed for clone R12, with a membranous staining pattern of lymphocyte subsets (datasheet). Figure A of the datasheet, shown as figure 03 in the gallery, presents normal human tonsil with numerous CD112R/PVRIG-positive lymphocytes in brightfield IHC; the multiplex figures of the gallery include CD112R × CD8 and CD112R × FOXP3 staining of normal human tonsil. In the Jansen thesis (2020), DIA-R12 showed a consistently membranous pattern; the published studies report CD112R signal on CD8+, CD4+, FOXP3+ and CD56+ lymphocytes.

What evidence supports the characterization and tissue staining of clone R12?

ONCOdianova product information (datasheet reactivity text, ONCOdianova product documentation and gallery) states that clone R12 was validated for sensitivity and specificity on large quantities of normal and tumor tissues, displays no background in non-lymphoid cells and in epithelial cells, and combines high specificity with an inherently high signal-to-noise ratio, which makes it well suited for multiplex IHC studies of CD112R in human tissue. Beyond the manufacturer, the Jansen thesis (2020) reports that the specificity of DIA-R12 was examined by Western blot, ELISA and pre-absorption with CD112R protein before multiplex use, and Yang et al. 2023 used DIA-R12 in a peer-reviewed multiplex panel. Brightfield IHC, fluorescence multiplex IHC and Western blot are the documented applications.

Is Western blot a documented application of R12?

Yes. ONCOdianova's original R12 product information lists Western blot among the antibody's applications, alongside immunohistochemistry on human FFPE sections. In addition, the doctoral thesis of Jansen (2020, University of Hamburg) reports that the specificity of DIA-R12 was examined by Western blot, ELISA and pre-absorption with CD112R protein.

Is R12 suitable for automated IHC platforms?

The datasheet states that the antibody is suited for immunohistochemical staining on automated platforms, and the ONCOdianova product documentation describes its manual protocol as applicable to automated staining procedures. The documented conditions — heat-induced epitope retrieval at pH 7.8 and 1:100 to 1:200 for polymer-based detection — are the starting point for establishing R12 on a laboratory's own platform.

P·11 Documents

Ordering and catalogue information.

Datasheet

Datasheet_DIA-R12_anti-CD112R

Product data, instructions for use, storage, general references and the tonsil reference images (figures A and B).

Download PDF ↓

PDF · version 25.02.2025/04 · 267 KB

IHC gallery

CD112R IHC gallery

The ONCOdianova “CD112R IHC-Gallery” is presented in section P·04 of this page with 15 figures and their legends.

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15 figures · brightfield and multiplex

Safety

MSDS

Material safety data sheet (ONCOdianova, all lyophilized antibodies).

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PDF · V02 · 216 KB

Ordering. ONCOdianova products are purchased through our distribution partner BIOZOL Diagnostica Vertrieb GmbH, Oehleckerring 11–13, 22419 Hamburg, Germany — order requests by e-mail: order@biozol.de (CC info@oncodianova.com). See also order information.

The technical data on this page are taken from datasheet DIA-R12 (version 25 Feb 2025/04; product content identical to V03, 22 Feb 2022). Related: TG1 (anti-TIGIT) · TG2 (anti-TIGIT) · KK3 (CD73/NT5E) · TC8 (CD8) · FX3 (FOXP3).

For Research Use Only. Not for use in diagnostic procedures.