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HAVCR2 / Hepatitis A virus cellular receptor 2 · IHC design guide
Design Immunohistochemistry for HAVCR2
Plan HAVCR2 IHC around cytoplasmic staining in a subset of lymphoid cells (HPA tissue IHC) and its expected membrane location (UniProt). This guide covers fixation, controls, and interpretation of paraffin-section staining.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
HAVCR2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasm observed in some lymphoid cells (HPA tissue IHC); cell membrane expected (UniProt) | |
| Staining pattern | Subset of lymphoid cells with cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+3 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00657) | |
| Caveat | Presumed off-target staining may mimic positive cells (HPA tissue IHC) | |
| Regulation | Treg expression follows TCR stimulation (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope against extracellular/cytoplasmic topology (UniProt) |
Section 1
Recommended HAVCR2 IHC & IF Protocols
The catalog antibody protocol is accompanied by published IHC methods for paraffin-embedded tissues (PMC9751455) and human breast cancer FFPE tissue (PMC13397360).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A00657); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-HAVCR2, 2 μg/mL (datasheet A00657) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | HAVCR2-positive staining in proximal tubules (microvilli) of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in a subset of lymphoid cells. No signal in the no-primary control. |
💡Decision noteFor this membrane antigen (UniProt Q8TDQ0 topology), start with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule).
Section 2
What Is the Expected HAVCR2 Staining Pattern?
HAVCR2 is a cell membrane protein with an extracellular region at residues 22–202 and a cytoplasmic region at 224–301 (UniProt Q8TDQ0 topology). In IHC, expect strong proximal-tubule microvillus staining in kidney and staining in a subset of lymphoid cells; HPA also describes cytoplasmic staining in those lymphoid cells (HPA tissue IHC). HPA rates its IHC evidence Enhanced, while noting medium agreement with RNA data and disregarded presumed off-target staining (HPA tissue IHC).
| Strong staining along kidney proximal-tubule microvilli, with scattered positive non-germinal-center lymphoid cells. | This fits HPA's High kidney microvillus signal and Medium lymph-node and tonsil signal (HPA tissue IHC). Assess each named cell population separately: the lymphoid pattern is a subset, so a positive section need not show every lymphoid cell stained (HPA tissue IHC). |
| Predominantly nuclear staining, especially where expected microvillus or lymphoid staining is absent. | A nuclear-dominant pattern conflicts with the reported membrane location and transmembrane topology (UniProt Q8TDQ0 topology; HPA subcellular). Review localisation against a positive kidney section and a no-primary control before scoring it as HAVCR2 (general IHC practice). HPA's reported cytoplasmic lymphoid staining should not be mistaken for nuclear staining (HPA tissue IHC). |
| Convincing staining in adipocytes or other cells HPA lists as not detected. | HPA reports adipocytes as Not detected in adipose tissue and breast (HPA tissue IHC). Check cell identification and control staining; cross-reactivity or detection-system activity can mimic a positive result (general IHC practice). HPA also reports presumed off-target binding in its overall reliability assessment (HPA tissue IHC). |
| Diffuse chromogen covers cells and extracellular areas without a readable cell pattern. | A uniform haze cannot establish the reported proximal-tubule or subset-of-lymphoid-cells pattern (HPA tissue IHC). Inspect a no-primary section for detection background and review blocking, washes and chromogen development before interpreting individual cells (general IHC practice). |
| No signal in an adequately sampled kidney section, including proximal-tubule microvilli. | This conflicts with HPA's High proximal-tubule microvillus staining (HPA tissue IHC). First assess section quality, tissue identification and assay controls, then review the antibody dilution, retrieval and detection steps against the laboratory's validated IHC procedure (general IHC practice). |
💡Expected HAVCR2 appearanceCall the result positive when kidney proximal-tubule microvilli stain strongly and/or a subset of non-germinal-center lymphoid cells stains at a moderate level; nuclear-dominant or indiscriminate all-cell colour warrants control review (HPA tissue IHC; UniProt Q8TDQ0 topology; general IHC practice).
| Membrane topology and compartment | HAVCR2 spans residues 203–223, with extracellular and cytoplasmic portions on opposite sides of the membrane (UniProt Q8TDQ0 topology). The catalog antibody's epitope is not supplied here, so topology alone does not establish which portion it recognizes or which retrieval condition works. |
| Cell population and tissue choice | Kidney proximal-tubule microvilli are High; duodenal Paneth cells and lymph-node and tonsil non-germinal-center cells are Medium (HPA tissue IHC). Lung macrophages, small-intestinal Paneth cells and appendix non-germinal-center cells are listed as Low, making a faint result less decisive there (HPA tissue IHC). |
| Evidence and antibody validation | HPA calls the tissue IHC result Enhanced and lists CAB026003 as IHC Enhanced (HPA tissue IHC; HPA antibodies). Its reliability note also reports medium staining–RNA consistency and presumed off-target binding that was disregarded; unusual positives therefore need control review (HPA tissue IHC; general IHC practice). |
| IF/ICC Q&A: Where should signal appear? | HPA's subcellular summary is Membrane, and UniProt lists cell membrane and cell junction, including the immunological synapse between CD8+ T cells and targets (HPA subcellular; UniProt Q8TDQ0 subcellular). HPA supplies no ICC-IF cell-line images here, so this record cannot confirm a cell-line-specific IF pattern (HPA subcellular). |
| Protein forms and modifications | UniProt lists two isoforms, a cleaved signal peptide at residues 1–21, and glycosylation sites at 145 and 172 (UniProt Q8TDQ0). Without epitope information, these annotations do not establish isoform discrimination, staining intensity or a HAVCR2-specific retrieval effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney proximal tubules are blank. | The result disagrees with HPA's High microvillus staining; the specific failed assay step cannot be inferred from the image alone (HPA tissue IHC; general IHC practice). | Confirm proximal tubules and tissue integrity, then compare a positive control and review antibody dilution, retrieval, incubation and detection against the validated IHC procedure (general IHC practice). No HAVCR2-specific retrieval response is supplied. |
| Lymph node or tonsil looks negative at low magnification. | HPA places the Medium signal in non-germinal-center cells and describes expression in a subset of lymphoid cells, so sparse positives can be missed (HPA tissue IHC). | Inspect representative non-germinal-center areas at higher magnification and score cells individually; use the kidney microvillus pattern as a separate reference for assay performance (HPA tissue IHC; general IHC practice). |
| Nuclei carry the strongest signal. | Nuclear-dominant staining does not fit the membrane localisation or topology reported for HAVCR2 (HPA subcellular; UniProt Q8TDQ0 topology). Background or nonspecific detection is a possible assay explanation (general IHC practice). | Compare with a no-primary section, reassess counterstain versus chromogen, and verify whether the expected kidney or lymphoid compartments stain (general IHC practice; HPA tissue IHC). |
| Adipocytes stain strongly. | HPA reports adipocytes as Not detected in adipose tissue and breast; its reliability note also flags presumed off-target binding (HPA tissue IHC). Endogenous detection activity is another possible explanation (general IHC practice). | Verify the stained cell type, examine a no-primary control and review blocking and detection controls before assigning HAVCR2 positivity (general IHC practice). |
| Colour spreads across most of the section. | A widespread, poorly localised signal does not match HPA's specified microvillus and subset-of-lymphoid-cells observations (HPA tissue IHC). Excess detection background or endogenous activity can produce this appearance (general IHC practice). | Check a no-primary control, review endogenous-enzyme blocking where applicable, and assess washes and chromogen development; interpret target staining only after cellular boundaries are readable (general IHC practice). |
Experiment evidence
IHC & IF images showing HAVCR2 staining
A00657 · IHCImmunohistochemistry of TIM-3 in human spleen tissue with TIM-3 antibody at 2 μg/ml.
A00657 · IFImmunofluorescence of TIM-3 in human spleen tissue with TIM-3 antibody at 20 μg/ml. Red: TIM-3 Antibody (A00657) Blue: DAPI staining
M00657 · IHCImmunohistochemistry of TIM-3 in human colon carcinoma tissue with TIM-3 antibody at 5 μg/mL.
M00657 · IFImmunocytochemistry of TIM-3 in transfected HEK293 cells with TIM-3 antibody at 1 μg/mL. Lower left: Immunocytochemistry in transfected HEK2
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000135077-HAVCR2/tissue/cerebral+cortex#imid_6423856
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000135077-HAVCR2/tissue/colon#imid_6423782
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000135077-HAVCR2/tissue/liver#imid_6423783
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000135077-HAVCR2/tissue/kidney#imid_6423784
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for HAVCR2 IHC & IF
🧪Run kidney first: proximal tubule microvilli should stain (HPA: High in kidney proximal tubules). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); neighboring cells on the kidney slide should lack the distinct microvillar staining pattern and show only low background.
Positive control tissue: Kidney (Proximal tubules (microvilli), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for HAVCR2; derive a cell-line control from the positive tissue's cell type (Proximal tubules (microvilli)) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and immunoglobulin class; use HAVCR2 knockout material or a peptide-block control, if available, to assess specificity. Quench endogenous peroxidase for chromogenic detection and check kidney sections for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence, so assess antigen retrieval empirically on paraffin sections. The selected A00657 spleen IHC caption gives 2 µg/ml but does not report a fixative (A00657 caption: fixative not stated). The supplied evidence does not establish whether frozen sections or IF/ICC are easier for HAVCR2; kidney tubules warrant particular attention to endogenous biotin in biotin-based IHC and autofluorescence if IF is evaluated (standard IHC/IF practice).
HPA tissue IHC evidence for HAVCR2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Kidney | Proximal tubules (microvilli) | High | Protein (IHC) | HPA → |
| Duodenum | Paneth cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced HAVCR2 IHC Tips
Troubleshoot HAVCR2 chromogenic IHC in paraffin sections by checking retrieval, cell identity, staining compartment, and assay controls before interpreting signal.
What retrieval conditions should I try first for weak HAVCR2 staining?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page IHC retrieval setting). Allow sections to cool in the buffer, then compare retrieved and unretrieved sections with the same antibody concentration and detection conditions (standard IHC practice). Evaluate staining in the cell population of interest, because HAVCR2 is a membrane protein also reported in a subset of lymphoid cells with cytoplasmic staining (UniProt Q8TDQ0 subcellular location; HPA tissue IHC profile). If signal remains weak, test a second retrieval condition as a fallback on matched sections, checking whether any gain in signal also raises background (standard IHC practice).
How can I assess whether fixation is causing weak HAVCR2 IHC?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the spleen IHC caption for A00657 does not state its fixative (A00657 tissue-IHC caption). Record the fixative, fixation duration, processing history, and section age for each specimen before comparing staining (standard IHC practice). Run matched sections through the same citrate pH 6.0 retrieval at 95–98 °C for 20 minutes, then hold antibody and detection conditions constant (page IHC retrieval setting; standard IHC practice). If specimens with different processing histories stain differently, repeat the comparison with a consistently processed control; the difference alone does not establish a HAVCR2-specific fixation effect (standard IHC practice).
Should HAVCR2 IHC appear on the membrane or in the cytoplasm?
Expect cell-surface or junction-associated signal from the annotated protein location, including localisation at the immunological synapse between CD8+ T cells and target cells (UniProt Q8TDQ0 subcellular location). Its extracellular region spans residues 22–202, its transmembrane segment 203–223, and its cytoplasmic region 224–301 (UniProt Q8TDQ0 topology). HPA also reports cytoplasmic staining in a subset of lymphoid cells, so document membrane and cytoplasmic patterns separately rather than merging them into one score (HPA tissue IHC profile; standard IHC practice). Compare morphology and cell identity across serial sections before assigning punctate or diffuse chromogen to HAVCR2 (standard IHC practice).
Could epitope position or isoforms explain discordant HAVCR2 staining?
First establish whether the catalog antibody recognises an extracellular or cytoplasmic epitope; the supplied A00657 caption does not identify its epitope (A00657 tissue-IHC caption). HAVCR2 has 2 annotated isoforms, but the supplied record does not show which sequence differences each contains, so isoform recognition cannot be inferred here (UniProt Q8TDQ0 isoforms). An extracellular epitope may lie near the Ig-like V-type domain at residues 22–124 or glycosylation sites at 145 and 172 if its mapped sequence overlaps them (UniProt Q8TDQ0 domains and glycosylation). Compare antibodies with documented, distinct epitopes on serial sections before interpreting discordant patterns as isoform-specific expression (standard IHC practice).
How should I investigate a HAVCR2 IHC pattern with multiplex IF?
Use the companion IF/ICC assay to pair HAVCR2 with a marker identifying the cells being scored, such as a validated macrophage marker when examining the lung macrophages listed by HPA (HPA tissue IHC low-expression list; standard IF practice). Choose fluorophores after imaging an unstained section, placing the weaker signal in a channel with low tissue autofluorescence (standard IF practice). For an extracellular epitope, compare staining with minimal permeabilisation; for a cytoplasmic epitope, test controlled permeabilisation, because HAVCR2 spans extracellular residues 22–202 and cytoplasmic residues 224–301 (UniProt Q8TDQ0 topology; standard IF practice). Confirm cell identity and compartment against the chromogenic sections without treating IF signal alone as proof of specificity (standard IHC/IF practice).
How do I distinguish HAVCR2 staining from nonspecific chromogenic background?
Include a no-primary control and inspect whether staining persists after the primary antibody is omitted; persistent chromogen implicates the detection workflow (standard IHC practice). Check the peroxidase block and inspect blood-rich or damaged areas separately when using peroxidase-based detection (standard IHC practice). Titrate the antibody around a documented starting point only when the same catalog antibody is used: A00657 was shown in human spleen IHC at 2 µg/ml (A00657 tissue-IHC caption). Because HPA reports presumed off-target binding in its tissue assessment, require plausible cell identity and compartment as well as a clean control before calling diffuse staining positive (HPA tissue IHC reliability description; UniProt Q8TDQ0 subcellular location).
What should I score when HAVCR2-positive cells are sparse? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and cell population before scoring, then report the percentage of positive eligible cells and an intensity-weighted H-score for that population (standard IHC practice). For scattered immune cells, also report positive-cell density per mm² of viable tissue, normalised to the measured viable area (standard IHC practice). Keep membrane and cytoplasmic scores separate because HAVCR2 is annotated at the membrane while HPA reports cytoplasmic staining in a subset of lymphoid cells (UniProt Q8TDQ0 subcellular location; HPA tissue IHC profile). Apply the same threshold, counterstain review, and exclusion rules across specimens; record excluded edges, folds, and necrotic regions (standard IHC practice).
When should I doubt a positive HAVCR2 IHC result?
Question a signal dominated by nuclei or by a cell type that lacks corroborating morphology, because HAVCR2 is annotated at the cell membrane and cell junction (UniProt Q8TDQ0 subcellular location; standard IHC practice). Compare the pattern with HPA’s reported high staining in kidney proximal-tubule microvilli and medium staining in lymph-node non-germinal-center cells, while recognising its warning about presumed off-target binding (HPA tissue IHC positive list and reliability description). Exclude section edges and necrotic areas from interpretation, and investigate chromogen in the no-primary control as possible endogenous enzyme or detection background (standard IHC practice). Call a result credible only when cellular location, cell identity, and controls agree across comparable sections (standard IHC practice).
Boster reagents
Best HAVCR2 / Hepatitis A virus cellular receptor 2 IHC Antibodies
The catalog includes human-reactive anti-HAVCR2 antibodies with IHC images from spleen and colon carcinoma tissue and IF images from spleen tissue and transfected HEK293 cells (catalog reactivity and image captions).
Real IHC data
Anti-TIM-3 HAVCR2 Antibody
Cat # A00657
Real IHC data
Anti-TIM3 HAVCR2 Monoclonal Antibody [10C10]
Cat # M00657
A00657 has an IHC-P image from human spleen at 2 μg/mL and an IF image from human spleen (catalog applications and image captions). M00657 has an IHC-P image from human colon carcinoma at 5 μg/mL, plus IF and ICC images from transfected HEK293 cells (catalog applications and image captions).
Which to pick: For paraffin-section IHC, choose A00657 for human spleen or M00657 for human colon carcinoma: each has its own IHC-P listing and tissue image, and neither image caption reports the fixative (catalog applications and respective IHC image captions). For IF or ICC, M00657 is the mouse monoclonal with both applications listed; A00657 is a rabbit antibody with IF listed and a human spleen IF image (catalog host, clone, applications and IF image caption). For work spanning human, mouse and rat, consider A00657-5, a rabbit polyclonal listed for IHC and IF in those species; the payload provides no IHC or IF image for it (catalog reactivity, applications, dilution_raw and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TDQ0 (HAVR2_HUMAN, Hepatitis A virus cellular receptor 2).
- Human Protein Atlas. HAVCR2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. HAVCR2 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. HAVCR2 antibody validation summary (1 antibodies).
- Integrating cell interaction with transcription factors to obtain a robust gene panel for prognostic prediction and therapies in cholangiocarcinoma. Frontiers in genetics 2022 — PMC9748417.
- Differential expression of HAVCR2 gene in pan-cancer: A potential biomarker for survival and immunotherapy. Frontiers in genetics 2022 — PMC9445440.
- Weighted gene co-expression network reveals driver genes contributing to phenotypes of anaplastic thyroid carcinoma and immune checkpoint identification for therapeutic targets. Frontiers in oncology 2022 — PMC9751455.
- Tumour-Derived sEVs Promote Triple-Negative Breast Cancer Progression Associated with HAVCR2 Upregulation in Macrophages. Oncology research 2026 — PMC13397360.
- PubMed PMID:11823861 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for HAVCR2 (Hepatitis A virus cellular receptor 2). Expected localisation Cytoplasm observed in some lymphoid cells (HPA tissue IHC); cell membrane expected (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A00657 at 2 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Proximal tubules (microvilli)); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A00657); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-HAVCR2, 2 μg/mL (datasheet A00657); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00657); Caveat: Presumed off-target staining may mimic positive cells (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope against extracellular/cytoplasmic topology (UniProt); Regulation: Treg expression follows TCR stimulation (UniProt). Evidence: UniProt Q8TDQ0, HPA tissue IHC · Enhanced, HPA ICC-IF · Membrane, Datasheet A00657, PMC9751455, PMC13397360. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/HAVCR2/immunohistochemistry-ihc-guide.