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VIPR2 / Vasoactive intestinal polypeptide receptor 2 · IHC design guide
Design Immunohistochemistry for VIPR2
Plan chromogenic VIPR2 IHC on paraffin sections using membranous staining as the expected tissue pattern (HPA tissue IHC). Compare high-staining bronchial basal cells with glandular cells of the adrenal gland, where staining was not detected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
VIPR2 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 | Membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Membranous staining in pneumocytes, smooth muscle and endothelium (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03768-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Adrenal gland+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium consistency; verify specificity (HPA tissue IHC) | |
| Regulation | CD4+ T cells express VIPR2; CD8+ T cells do not (UniProt) | |
| Isoform / epitope | 2 isoforms; check extracellular vs cytoplasmic epitope (UniProt) |
Section 1
Recommended VIPR2 IHC & IF Protocols
The catalog antibody provides the paraffin-section IHC protocol (datasheet A03768-1). The published option describes chromogenic IHC on free-floating brain sections (PMC4757782).
| Sample | Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A03768-1) |
| Fixation | Image fixative and duration unreported (datasheet A03768-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03768-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03768-1) |
| Primary antibody | Rabbit anti-VIPR2, 2-5 μg/ml (datasheet A03768-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03768-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03768-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | VIPR2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Membranous expression in most tissues, including pneumocytes, smooth muscle cells and endothelial cells. No signal in the no-primary control. |
💡Decision noteStart paraffin sections with heat-mediated EDTA pH 8.0 retrieval (datasheet A03768-1). The published free-floating protocol does not specify retrieval (PMC4757782).
Section 2
What Is the Expected VIPR2 Staining Pattern?
VIPR2 is a cell-membrane receptor with 7 transmembrane segments (UniProt P41587 topology). In paraffin-section IHC, expect predominantly membranous staining in selected cells, including pneumocytes, smooth muscle cells, endothelial cells, adipocytes, bronchial basal cells and glomerular cells (HPA tissue IHC). HPA rates its tissue IHC evidence Approved, with medium consistency between antibody staining and RNA expression; interpret individual positive cells and compartments with that limitation in mind (HPA tissue IHC).
| Clear membrane-associated staining in adipocytes or bronchial basal cells, with limited surrounding background. | This fits the expected compartment (UniProt P41587 topology; HPA tissue IHC). HPA reports High staining in adipocytes in adipose tissue and breast, and in bronchial basal cells (HPA tissue IHC). Compare the relevant cell population, rather than calling an entire mixed tissue uniformly positive. |
| Strong nuclear staining, or uniform cytoplasmic color without a discernible membrane pattern. | That distribution does not match the annotated cell-membrane location or the predominantly membranous tissue pattern (UniProt P41587; HPA tissue IHC). Treat it as suspect staining; review morphology, the negative control and detection conditions before assigning it to VIPR2 (general IHC practice). |
| Prominent staining in adrenal glandular cells or bone-marrow hematopoietic cells. | HPA reports VIPR2 staining as Not detected in those specified cell populations (HPA tissue IHC). An unexpected signal may reflect cross-reactivity or endogenous detection activity; check a no-primary control and whether the signal follows the same compartmental pattern as the positive tissue (general IHC practice). |
| Diffuse color covers many structures, obscures cell boundaries or appears on the no-primary control. | This is background rather than an interpretable membrane pattern (general IHC practice; HPA tissue IHC profile). Examine blocking, washing, detection reagent and chromogen development; use the no-primary control to locate signal arising after the primary-antibody step (general IHC practice). |
| No convincing membrane signal in an expected positive cell population. | First confirm that the section contains the intended cells: HPA reports High staining in bronchial basal cells, glomerular cells and placental decidual cells (HPA tissue IHC). If present, review the IHC-validated antibody, retrieval and detection workflow (general IHC practice). HPA approval does not guarantee staining in every specimen (HPA tissue IHC reliability). |
💡Expected VIPR2 appearanceCall a positive result when the appropriate cells show discernible, predominantly membranous staining at an intensity consistent with their HPA category—for example, High in bronchial basal cells—while broad nuclear or diffuse background staining remains suspect (HPA tissue IHC; UniProt P41587 topology; general IHC practice).
| Membrane topology and epitope access (UniProt P41587 topology). | VIPR2 has extracellular and cytoplasmic regions around 7 transmembrane segments (UniProt P41587 topology). The antibody epitope is not supplied here, so topology alone cannot specify an antigen-retrieval condition or predict staining strength in paraffin sections. |
| Signal peptide and mature chain (UniProt P41587 processing). | The annotated signal peptide spans residues 1–23; the receptor chain spans 24–438 (UniProt P41587 processing). Without the antibody epitope, this record cannot establish whether processing affects recognition in IHC. |
| Glycosylation and isoforms (UniProt P41587). | UniProt lists 3 glycosylation sites at residues 58, 88 and 92, and 2 isoforms (UniProt P41587). These annotations do not identify which form the catalog antibody detects or explain a particular IHC intensity. |
| Tissue and cell-population choice (HPA tissue IHC). | HPA reports High staining in glomerular cells and placental decidual cells, Medium in endometrial stromal cells, and Not detected in adrenal glandular cells (HPA tissue IHC). Score the annotated cell type; a whole-section average can conceal the relevant pattern. |
| IF/ICC Q: Should focal-adhesion staining define a positive paraffin IHC result? | A: No. HPA reports focal adhesion sites in ICC-IF, while its tissue IHC profile is membranous (HPA subcellular ICC-IF; HPA tissue IHC). The listed antibody's ICC status is Uncertain (HPA antibodies); assess IF/ICC on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue has no visible staining. | The sampled field may lack the relevant cells, or the IHC workflow may have failed (general IHC practice). HPA's High label applies to specified cells, such as bronchial basal cells, rather than every cell in bronchus (HPA tissue IHC). | Locate the annotated cells on the counterstained section, then check the IHC-validated antibody, antigen-retrieval run, primary incubation and detection controls (general IHC practice). Avoid inferring target-specific fixation sensitivity from this result. |
| Signal is widespread in a tissue expected to be weak or negative. | HPA reports Not detected in adrenal glandular cells and Low in bladder urothelial cells (HPA tissue IHC). Broad signal there raises concern for nonspecific primary binding or endogenous detection activity (general IHC practice). | Compare cell identity and compartment with a high-staining reference section; inspect no-primary and detection controls, blocking and chromogen development (general IHC practice). Treat HPA's 'Not detected' category as an observation, not proof of biological absence. |
| Only a nuclear or uniformly cytoplasmic pattern appears. | The pattern conflicts with VIPR2's cell-membrane annotation and HPA's membranous tissue profile (UniProt P41587; HPA tissue IHC). Background or antibody cross-reactivity is possible (general IHC practice). | Recheck the cell boundaries, compare a known HPA-high cell population, and review no-primary controls and detection conditions before scoring the section positive (HPA tissue IHC; general IHC practice). |
| Staining obscures the membranes that need scoring. | Excess chromogen development, insufficient washing or detection background can reduce contrast (general IHC practice). This prevents assessment of the expected membranous pattern (HPA tissue IHC). | Review development time and wash steps, then inspect the no-primary control. Score only cells whose boundaries and compartment remain distinguishable (general IHC practice). |
| A medium or low HPA category is being called a failed experiment. | HPA reports Medium staining in endometrial stromal and esophageal squamous cells, and Low staining in oral squamous and bladder urothelial cells (HPA tissue IHC). These categories differ from its high-staining examples. | Compare like cell types and use a separate HPA-high reference population to check assay performance; retain the reported intensity category when interpreting the test tissue (HPA tissue IHC; general IHC practice). |
| Two sections or antibodies give different-looking patterns. | HPA rates the tissue IHC evidence Approved but describes only medium consistency with RNA expression, and the listed antibody is Approved for IHC (HPA tissue IHC; HPA antibodies). Those ratings do not resolve every discordant cell-level result. | Compare the same annotated cell populations, compartment, controls and staining conditions across sections; report unresolved disagreement instead of treating one unexpected pattern as confirmed VIPR2 (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing VIPR2 staining
A03768-1 · IHCIHC analysis of VPAC2/VIPR2 using anti-VPAC2/VIPR2 antibody (A03768-1). VPAC2/VIPR2 was detected in a paraffin-embedded section of human pro
A03768-1 · IHCIHC analysis of VPAC2/VIPR2 using anti-VPAC2/VIPR2 antibody (A03768-1). VPAC2/VIPR2 was detected in a paraffin-embedded section of human lun
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106018-VIPR2/tissue/cerebral+cortex#imid_17301539
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106018-VIPR2/tissue/colon#imid_17301473
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106018-VIPR2/tissue/liver#imid_17301482
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106018-VIPR2/tissue/kidney#imid_17301467
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA062707; green = VIPR2, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA062707; green = VIPR2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for VIPR2 IHC & IF
🧪Run adipose tissue first and score adipocytes, which show high VIPR2 staining (HPA: High in adipocytes). Use adrenal glandular cells as the negative tissue (HPA: Not detected in adrenal glandular cells); on the adipose slide, unstained neighboring cells can serve as a background reference, but their VIPR2-negative status is unverified (HPA: adipocyte-specific row).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show VIPR2 in HeLa, SH-SY5Y, U2OS, with annotated localisation: Focal adhesion sites (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality (standard IHC practice). Confirm specificity with VIPR2 knockout tissue or a validated peptide-block control, and quench endogenous peroxidase before chromogenic detection in adipose sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected paraffin-section caption does not state a fixative (selected-SKU tissue-IHC caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required; whether frozen sections or IF are easier is unreported (selected-SKU tissue-IHC caption: EDTA retrieval). In adipose sections, check lipid-rich areas for staining artefacts when scoring adipocytes (standard IHC practice; HPA: High in adipocytes).
HPA tissue IHC evidence for VIPR2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Basal cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in glomeruli | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced VIPR2 IHC Tips
Troubleshoot VIPR2 staining in paraffin sections by checking retrieval, cell identity and membrane localisation against the catalog antibody’s tissue image and reference patterns (datasheet A03768-1; HPA tissue IHC; UniProt P41587).
What should I change when VIPR2 staining is weak after antigen retrieval?
Start with heat-mediated EDTA retrieval at pH 8.0 for paraffin sections (datasheet A03768-1). The catalog antibody produced a tissue image using that condition, followed by 2 μg/ml primary antibody overnight at 4°C; the caption gives no retrieval time, so record the time used in your run (datasheet A03768-1). Compare adjacent sections retrieved for different durations while holding antibody concentration and DAB development constant (standard IHC practice). Check whether stronger staining outlines cells in the expected compartment rather than appearing uniformly across the section (UniProt P41587 localisation; standard IHC practice). Include a no-primary control to distinguish retrieval-associated background from antibody signal (standard IHC practice).
How can I assess whether fixation caused weak VIPR2 IHC staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A03768-1). Record the fixative, fixation duration and processing history for each specimen before comparing staining intensities (standard IHC practice). On adjacent sections from the same block, keep EDTA at pH 8.0, primary antibody at 2 μg/ml overnight at 4°C, and DAB development consistent while checking other sources of variation (datasheet A03768-1; standard IHC practice). Include a tissue region with a documented cell-level staining pattern as a reference, such as glomerular cells in kidney (HPA: High in kidney glomerular cells). If reference staining also fails, investigate processing records and detection controls before attributing the result to VIPR2 biology (standard IHC practice).
Where should convincing VIPR2 staining appear in a tissue section?
Prioritise cell-associated membrane staining because VIPR2 is annotated as a cell-membrane receptor with 7 transmembrane segments (UniProt P41587 topology and localisation). HPA describes membranous tissue expression, including pneumocytes, smooth muscle cells and endothelial cells, while its cell-imaging record places VIPR2 at focal adhesion sites (HPA tissue IHC; HPA subcellular). Examine cell boundaries at high magnification and compare them with a counterstain before assigning a stained structure to a cell type (standard IHC practice). A solely nuclear or diffuse extracellular DAB pattern conflicts with those localisation records and needs investigation (UniProt P41587 localisation; HPA tissue IHC; standard IHC practice). Use cell identity and controls alongside compartment when judging specificity (standard IHC practice).
Can this IHC stain distinguish VIPR2 isoforms or reveal an inaccessible epitope?
VIPR2 has 2 annotated isoforms, but the supplied antibody caption gives no epitope or isoform-specificity information (UniProt P41587 isoforms; datasheet A03768-1). The mature chain spans residues 24–438, with an extracellular region at 24–124 and a cytoplasmic tail at 379–438 (UniProt P41587 processing and topology). Its extracellular region contains glycosylation sites at 58, 88 and 92 (UniProt P41587 glycosylation). Because the recognized epitope is unspecified, neither glycosylation nor membrane orientation establishes why a particular section stains weakly (datasheet A03768-1; UniProt P41587 topology and glycosylation). Treat staining as VIPR2-associated rather than isoform-specific, and seek epitope documentation before making an isoform claim (standard IHC interpretation; datasheet A03768-1).
How should I adapt the VIPR2 IHC finding for multiplex IF?
Use the chromogenic result as a localisation reference, then validate the antibody and detection conditions separately for IF; the supplied product image documents paraffin-section IHC with DAB (datasheet A03768-1; standard IF practice). Pair VIPR2 with a marker identifying the cell type being evaluated, such as a suitable endothelial marker when examining the endothelial pattern reported by HPA (HPA tissue IHC; standard IF practice). Select fluorophores and acquisition channels after inspecting unstained tissue for autofluorescence, and include single-colour controls for spectral overlap (standard IF practice). The antibody epitope is unspecified despite VIPR2 having extracellular and cytoplasmic regions, so establish whether permeabilisation is needed by comparing otherwise matched conditions (datasheet A03768-1; UniProt P41587 topology; standard IF practice).
What should I check when DAB obscures VIPR2 membrane staining?
Run no-primary and secondary-only controls to locate detection background before changing the primary antibody concentration (standard IHC practice). The selected image used 10% goat-serum blocking, 2 μg/ml rabbit primary overnight at 4°C, a peroxidase-conjugated secondary and DAB (datasheet A03768-1). Include a peroxidase-blocking step and inspect its control because endogenous enzyme activity can produce chromogenic signal (standard IHC practice). Check that wash steps remove unbound reagent, then compare DAB development times on adjacent sections without changing the retrieval condition (standard IHC practice; datasheet A03768-1). Score cell-boundary staining separately from pigment, tissue folds and section-edge deposits (UniProt P41587 localisation; standard IHC practice).
How should I quantify VIPR2 IHC across sections with different cell mixtures? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, because HPA reports differing VIPR2 staining levels among cell types (HPA tissue IHC; standard IHC practice). Record the percentage of eligible cells at each membrane-intensity grade, then calculate an H-score from 0–300, or report the percentage of positive eligible cells using a fixed threshold (standard IHC practice). For spatial comparisons, count positive cells per mm² within the same annotated compartment (standard IHC practice). Normalise cell-based scores to the number of eligible cells, and area-based densities to the analysed tissue area, excluding folds, necrosis and empty space (standard IHC practice). Keep retrieval, DAB development and image-analysis thresholds consistent across the comparison (standard IHC practice).
How can I distinguish true VIPR2 staining from section artefact?
Look for reproducible cell-associated membrane staining, consistent with VIPR2’s cell-membrane annotation and HPA’s tissue pattern (UniProt P41587 localisation; HPA tissue IHC). Interpret the stained cell type as well as the organ: HPA reports High staining in kidney glomerular cells but Not detected in adrenal glandular cells, so those observations do not describe every cell in either tissue (HPA tissue IHC). Review nuclear-only signal, section-edge accentuation, necrotic areas and staining present in a no-primary control as possible artefacts (UniProt P41587 localisation; standard IHC practice). Use peroxidase-blocking and detection controls to assess endogenous enzyme signal before calling weak DAB positive (standard IHC practice). HPA rates its tissue antibody data Approved with medium staining-to-RNA consistency, so corroborate consequential findings independently (HPA tissue IHC).
Boster reagents
Best VIPR2 / Vasoactive intestinal polypeptide receptor 2 IHC Antibodies
A03768-1 has real IHC data from paraffin sections of human prostate cancer, lung cancer and tonsil (IHC captions). No IF/ICC data are supplied (catalog).
Real IHC data
Anti-VPAC2/VIPR2 Antibody ®
Cat # A03768-1
A03768-1 is listed for IHC and has images from paraffin sections of human prostate cancer, lung cancer and tonsil (catalog; IHC captions). It is listed as reactive with human and monkey, but the supplied IHC images show human tissue only (catalog; IHC captions).
Which to pick: Choose the polyclonal A03768-1 for human paraffin-section IHC; its captions document EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody, while the catalog gives an IHC range of 2–5 μg/ml (catalog; IHC captions). Its paraffin-section captions do not report the fixative (IHC captions). No IF/ICC option is validated here; A03768-1 lists monkey reactivity, but the supplied IHC images are from human tissue only (catalog; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P41587 (VIPR2_HUMAN, Vasoactive intestinal polypeptide receptor 2).
- Human Protein Atlas. VIPR2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. VIPR2 subcellular location (ICC-IF): Localized to the focal adhesion sites..
- Human Protein Atlas. VIPR2 antibody validation summary (1 antibodies).
- A Distinct Subpopulation of Extended Amygdala Neurons Drives Food Intake. bioRxiv : the preprint server for biology 2025 — PMC12642667.
- A Distinct Subpopulation of Extended Amygdala Neurons Drives Food Intake. Biological psychiatry global open science 2026 — PMC13260202.
- Live imaging of altered period1 expression in the suprachiasmatic nuclei of Vipr2-/- mice. Journal of neurochemistry 2008 — PMC2658715.
- Gpr176 is a Gz-linked orphan G-protein-coupled receptor that sets the pace of circadian behaviour. Nature communications 2016 — PMC4757782.
- PubMed PMID:7811244 — UniProt-cited evidence.
- PubMed PMID:8938447 — UniProt-cited evidence.
- PubMed PMID:8933357 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for VIPR2 (Vasoactive intestinal polypeptide receptor 2). Expected localisation Membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03768-1). Validated IHC antibody: A03768-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet A03768-1); 2. Fixation: Image fixative and duration unreported (datasheet A03768-1); 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 retrieval: EDTA pH 8.0 (datasheet A03768-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03768-1); 8. Primary antibody: Rabbit anti-VIPR2, 2-5 μg/ml (datasheet A03768-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03768-1). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium consistency; verify specificity (HPA tissue IHC); Isoform / epitope: 2 isoforms; check extracellular vs cytoplasmic epitope (UniProt); Regulation: CD4+ T cells express VIPR2; CD8+ T cells do not (UniProt). Evidence: UniProt P41587, HPA tissue IHC · Approved, HPA ICC-IF · focal adhesion sites., Datasheet A03768-1, PMC4757782. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/VIPR2/immunohistochemistry-ihc-guide.