CAV2 / Caveolin-2 · IHC design guide

Design Immunohistochemistry for CAV2

Plan CAV2 chromogenic IHC in paraffin sections using cell type staining reported by HPA (HPA tissue IHC) and caveolar membrane localisation as a molecular expectation (UniProt). The guide covers fixation consistency, controls and scoring by cell type.

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for CAV2 (IHC for CAV2): expected localisation Caveolar membrane expected (UniProt); tissue compartment unreported (HPA tissue IHC), antibody A01574, validated IHC image, and IHC protocol steps
Printable CAV2 IHC protocol sheet — expected localisation Caveolar membrane expected (UniProt); tissue compartment unreported (HPA tissue IHC), antibody A01574, controls and protocol steps. Open the full CAV2 IHC guide →

CAV2 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 Caveolar membrane expected (UniProt); tissue compartment unreported (HPA tissue IHC)
Staining pattern Adipocytes, endothelium, pneumocytes and squamous basal cells (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet A01574)
Positive control ⓘ Adipose tissue+4 more · see all
Negative control ⓘ Appendix+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 Adipocytes can dominate staining in mixed tissue (HPA tissue IHC)
Regulation Insulin may enrich nuclear signal (UniProt)
Isoform / epitope Three isoforms; epitope differences unreported, both termini cytoplasmic (UniProt)
Section 1

Recommended CAV2 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: A01574) is accompanied by published CAV2 protocols for breast cancer tissue microarrays (PMC2480981) and head and neck squamous cell carcinoma sections (PMC9481523).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human rectal cancer tissue; fixative not specified (datasheet A01574)
FixationImage fixative and duration unreported (datasheet A01574); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet A01574); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A01574)
Primary antibodyRabbit anti-CAV2, 0.5-1μg/ml (datasheet A01574)
Primary incubationOvernight at 4 °C (datasheet A01574)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet A01574)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultCAV2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Distinct expression in adipocytes, endothelial cells, pneumocytes and basal cells of squamous epithelia. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A01574). Use each article’s stated retrieval conditions when reproducing its protocol (PMC2480981; PMC9481523).
Section 2

What Is the Expected CAV2 Staining Pattern?

CAV2 is a caveolar membrane protein with cytoplasmic-facing regions; UniProt also reports cytoplasmic, Golgi and conditional nuclear localization (UniProt P51636 topology and localization). In paraffin-section IHC, expect staining in adipocytes, endothelial cells, pneumocytes and basal cells of squamous epithelia (HPA tissue IHC: Enhanced reliability). Judge the pattern by the stained cell type as well as its compartment.

What am I looking at on my slide?
Distinct peripheral staining in adipocytes or endothelial cells, with little surrounding background.Consistent with CAV2 at caveolar membranes (UniProt P51636 localization) and the HPA tissue profile (HPA: adipocytes and endothelial cells). Assess intact cells and tissue architecture before scoring.
Predominantly nuclear staining throughout many cell types, without a convincing peripheral pattern.Investigate nonspecific signal or an imaging artefact. Nuclear CAV2 is reported under specific phosphorylation and insulin-linked conditions (UniProt P51636); routine nuclear staining alone does not establish that state.
Strong signal in bone-marrow hematopoietic cells or appendix glandular cells.These cell populations are listed as not detected (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic activity; inspect a secondary-only control and the expected positive cell population.
Uniform haze across cells, stroma and empty slide areas.This lacks the cell-selective pattern in the HPA profile (HPA tissue IHC). Check blocking, washes and detection controls as general IHC practice before assigning the haze to CAV2.
No signal in adipocytes of an otherwise interpretable adipose section.HPA reports high staining in adipocytes (HPA: adipose tissue). Review the catalog antibody's IHC-P instructions, antigen retrieval and detection controls as general IHC troubleshooting; absence alone does not identify the failed step.
💡Expected CAV2 appearanceCall positive when intact adipocytes or endothelial cells show distinct, appreciable peripheral staining (HPA: high in adipocytes; UniProt P51636: caveolar membrane); broad staining of cell populations listed as not detected is suspect (HPA tissue IHC).
How each factor affects the staining
Compartment and topologyCAV2 has cytoplasmic-facing regions and no annotated transmembrane segment (UniProt P51636 topology). Expect membrane-associated staining, while allowing reported cytoplasmic or Golgi signal (UniProt P51636 localization); a peripheral pattern need not outline every cell continuously.
Cell population within a sectionHPA reports high staining in adipocytes in adipose tissue and breast, and in Bowman's capsule cells in kidney (HPA tissue IHC). Its negative calls name specific cell populations; they do not make the entire organ a negative control.
Phosphorylation-dependent locationUniProt places Tyr-27-phosphorylated CAV2 in cytoplasm and plasma membrane, Ser-36-phosphorylated CAV2 in intracellular compartments, and Tyr-19-phosphorylated CAV2 in the nucleus after insulin-linked signaling (UniProt P51636). Total-CAV2 staining cannot identify a modification.
Antibody evidenceHPA lists IHC as Supported for HPA044810 and Enhanced for CAB013488 (HPA antibodies). The tissue profile also has Enhanced reliability (HPA tissue IHC). Apply the validation status to the antibody used; it does not validate every unexpected compartment.
IF/ICC Q&A: should vesicles appear?Yes: vesicles are the approved ICC-IF main location (HPA subcellular). This is a microscopy comparison, not an IHC-P protocol option. The paraffin-section interpretation should still use tissue-specific IHC evidence (HPA tissue IHC).
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Adipose positive control is blank.The missing signal could arise at retrieval, primary incubation or detection; HPA staining does not establish CAV2-specific fixation sensitivity (HPA: adipocytes High).Follow the catalog antibody's IHC-P instructions and check retrieval and detection with an established control (general IHC practice).
Only the nuclear compartment is strongly stained.Nuclear localization is conditional in the UniProt account (UniProt P51636); an isolated nuclear result cannot confirm the reported signaling state.Compare with a positive cell population and a secondary-only control; review nuclear counterstain and morphology (general IHC practice).
Hematopoietic or glandular cells stain strongly.HPA lists bone-marrow hematopoietic cells and appendix glandular cells as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible.Check a secondary-only section and the detection-system block, then compare the suspect cells with an HPA-positive cell population (general IHC practice).
All regions have brown haze.Diffuse staining does not match the distinct cellular profile (HPA tissue IHC); excess detection background is possible.Inspect no-primary controls, blocking and washes; adjust the IHC workflow only against a retained positive control (general IHC practice).
A kidney or breast section appears partly negative.HPA's high calls apply to Bowman's capsule cells in kidney and adipocytes in breast, not every cell in those sections (HPA tissue IHC).Score the named cell population separately using morphology; avoid calling the whole section positive or negative (HPA tissue IHC).
IHC and ICC-IF show different-looking patterns.HPA reports vesicles in ICC-IF and distinct positive tissue cell populations in IHC (HPA subcellular; HPA tissue IHC); the observations use different preparations.Interpret the paraffin section against its IHC cell profile; use the separate IF/ICC guide for fluorescence workflow (HPA tissue IHC; HPA subcellular).

Sample controls for CAV2 IHC & IF

🧪Run adipose tissue first and require staining in adipocytes (HPA: High in adipocytes); use appendix glandular cells as the tissue negative (HPA: Not detected in appendix glandular cells). On the positive slide, cells without specific signal should show only background chromogen; do not assume stromal or endothelial cells are internal negatives because CAV2 is expressed in fibroblasts and endothelial cells (UniProt P51636 tissue specificity).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: Appendix (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CAV2 in A-431, A-549, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and nonimmune rabbit IgG matched to the primary antibody concentration (A01574 caption: rabbit antibody at 1 μg/ml); confirm specificity with CAV2 knockout material or an immunizing-peptide block if available (standard IHC controls). For adipose tissue, quench endogenous peroxidase and check for endogenous biotin staining before interpreting the biotin–SABC/DAB signal (A01574 caption: biotinylated secondary and SABC/DAB detection; standard IHC practice).
⚠️Feasibility: The documented paraffin-section starting condition is heat retrieval in EDTA at pH 8.0, followed by 1 μg/ml primary antibody overnight at 4 °C; absolute retrieval dependency is unreported (A01574 caption). A target-specific fixation window and fixation effect are unreported, and the selected caption does not state the fixative (A01574 caption: fixative not stated). No matched evidence establishes that frozen sections or IF are easier; lipid extraction in processed adipose tissue can leave clear vacuoles and narrow cytoplasmic rims that complicate scoring (standard paraffin histology).

HPA tissue IHC evidence for CAV2

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes High Protein (IHC) HPA →
Breast Adipocytes High Protein (IHC) HPA →
Bronchus Respiratory epithelial cells High Protein (IHC) HPA →
Cervix Squamous epithelial cells High Protein (IHC) HPA →
Endometrium Cells in endometrial stroma High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Duodenum Glandular cells Not detected Protein (IHC) HPA →
Section 3

Advanced CAV2 IHC Tips

Troubleshoot CAV2 staining in paraffin sections by checking retrieval, cell identity and subcellular pattern before interpreting chromogenic signal.

Which retrieval conditions should I try first for CAV2 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01574). The selected tissue-IHC example used that retrieval before 10% goat-serum blocking and overnight incubation at 4°C with 1 µg/mL primary antibody (datasheet A01574). Keep retrieval conditions and section thickness consistent across comparison slides, and include a positive control with each run (standard IHC practice). If staining is weak, adjust heating duration in a small pilot series while holding antibody concentration and detection constant (standard IHC practice). Excessive heating can damage tissue morphology, so assess both signal and section integrity (standard IHC practice).
Does fixation explain weak CAV2 staining in my paraffin sections?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown for this antibody (datasheet A01574). Document the fixative, fixation duration and processing history for each specimen before comparing staining intensity (standard IHC practice). On matched sections, first reproduce EDTA retrieval at pH 8.0 and the reported 1 µg/mL overnight primary incubation at 4°C (datasheet A01574). If fixation histories differ, test representative specimens together using the same retrieval and detection run, with a positive control (standard IHC practice). Interpret any intensity difference as an experimental observation until matched processing establishes its cause (standard IHC practice).
Where should convincing CAV2 staining appear within a cell?
Assess membrane-associated and intracellular staining in the context of the cell type: CAV2 is annotated at caveolae, cell membrane, Golgi membrane, cytoplasm and nucleus (UniProt P51636 subcellular location). Its reference topology places residues 1–86 and 108–162 on the cytoplasmic side of the membrane (UniProt P51636 topology). Vesicular localisation has also been observed by ICC/IF (HPA subcellular). Nuclear signal needs additional scrutiny because the proposed insulin-responsive translocation of Tyr-19-phosphorylated CAV2 is annotated by similarity (UniProt P51636 subcellular location). Compare candidate staining with a positive-control section and a no-primary control before assigning a compartment (standard IHC practice).
Could isoforms or phosphorylation change which CAV2 pool my antibody detects?
CAV2 has 3 annotated isoforms—Alpha, Beta and C—so establish which sequence region the antibody recognises before claiming pan-isoform staining (UniProt P51636 isoforms; standard IHC practice). The supplied evidence does not map the catalog antibody’s epitope or establish its isoform coverage (datasheet A01574). The reference protein has cytoplasmic regions at residues 1–86 and 108–162, with annotated phosphorylation at Tyr-19, Ser-23, Tyr-27 and Ser-36 (UniProt P51636 topology and modified residues). If an epitope overlaps a modified region, compare validated reagents or controls before attributing a staining shift to phosphorylation (standard IHC practice). Report the antibody and epitope information available for the experiment alongside any isoform-level interpretation (standard IHC practice).
How should I investigate CAV2 localisation with IF alongside this IHC assay?
Use IF as a separate localisation experiment and multiplex CAV2 with a marker of the cell population being assessed, such as an endothelial-cell marker when evaluating vessels (UniProt P51636 tissue specificity; standard IF practice). Choose spectrally separated fluorophores and check unstained tissue for autofluorescence before selecting a far-red detection channel (standard IF practice). If the antibody epitope is confirmed to face the cytoplasm, evaluate permeabilisation against an otherwise matched control because the reference CAV2 topology is cytoplasmic on both sides of its membrane hairpin (UniProt P51636 topology; standard IF practice). HPA reports approved vesicular ICC/IF localisation, which offers a pattern to assess (HPA subcellular). The paraffin-IHC caption supplies no IF fixation or permeabilisation conditions (datasheet A01574).
How can I distinguish CAV2 signal from chromogenic background?
Run a no-primary control through the same secondary, complex and DAB steps to reveal detection-system background (standard IHC practice). The selected example used 10% goat-serum blocking, biotinylated goat anti-rabbit secondary for 30 minutes at 37°C, and a streptavidin–biotin complex with DAB (datasheet A01574). Include an endogenous-peroxidase block, and assess endogenous biotin when using a biotin-based system (standard IHC practice). If diffuse colour persists, review washing, primary concentration and DAB development time using matched sections (standard IHC practice). Compare the resulting pattern with expected endothelial or adipocyte staining before calling widespread colour CAV2 positive (HPA tissue IHC; standard IHC practice).
How should I score CAV2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; CAV2 staining is reported in adipocytes and endothelial cells, among other populations (HPA tissue IHC). For a specified population, record the percentage of positive viable cells and an intensity score of 0–3, then calculate an H-score of 0–300 when intensity distribution matters (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of evaluated tissue, with the sampled area and exclusion rules (standard IHC practice). Normalise cell counts to the relevant viable-cell denominator rather than total section area alone (standard IHC practice). Apply one threshold, counterstain and imaging setup across groups, and score membrane-associated and intracellular signal separately if both are analysed (standard IHC practice).
When is a CAV2-positive pattern credible rather than artefactual?
A credible result aligns cell identity, compartment and controls: CAV2 is reported in adipocytes and endothelial cells, and annotated at caveolar membranes and intracellular sites (HPA tissue IHC; UniProt P51636 subcellular location). In the selected paraffin example, rectal cancer tissue was stained, but the caption does not identify which cells produced the signal (datasheet A01574). Compare suspicious tumour-cell colour with adjacent vessels and a no-primary section before assigning it to malignant cells (standard IHC practice). Discount staining confined to section edges, necrotic areas or endogenous-enzyme-rich structures when the corresponding controls show similar colour (standard IHC practice). Treat isolated nuclear staining cautiously because the annotated Tyr-19-linked nuclear translocation is qualified as by similarity (UniProt P51636 subcellular location).
Boster reagents

Best CAV2 / Caveolin-2 IHC Antibodies

Catalog anti-CAV2 antibodies have IHC images from human, mouse and rat paraffin sections, plus IF images from human tissue and A549 cells (catalog image captions).

Real IHC data IHC analysis of Caveolin-2/CAV2 using anti Caveolin-2/CAV2 antibody (A01574). Caveolin-2/CAV2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Caveolin-2/CAV2 Antibody (A01574) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-Caveolin-2/CAV2 Antibody ®
Cat # A01574
Real IHC data IHC analysis of Caveolin-2 using anti-Caveolin-2 antibody (PB9166). Caveolin-2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Caveolin-2 Antibody (PB9166) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-Caveolin-2/CAV2 Antibody ®
Cat # PB9166
Real IHC data Anti-Caveolin-2 antibody, PA1540, IHC(P) IHC(P): Human Lung Cancer Tissue
Anti-Caveolin-2/CAV2 Antibody ®
Cat # PA1540
Real IHC data Immunohistochemical analysis of paraffin-embedded human colon, using Caveolin-2 Antibody.
Anti-Caveolin-2 CAV2 Rabbit Monoclonal Antibody
Cat # M01574

A01574 has IHC images from human rectal cancer and mouse and rat lung, plus IF images from human intestinal cancer and A549 cells; PB9166 has IHC images from human rectal cancer and rat lung, plus IF images from human rectal cancer and A549 cells (catalog image captions). PA1540 has IHC images from human lung cancer and rat cardiac muscle; M01574 has an IHC image from human colon (catalog image captions).

Which to pick: For paraffin-section IHC, A01574 or PB9166 has a documented starting workflow: EDTA retrieval at pH 8.0, 10% goat-serum block and 1 μg/ml primary overnight at 4°C, followed by biotin–streptavidin/DAB detection (A01574 and PB9166 IHC image captions); the fixative is unreported in both captions. For IF/ICC, PB9166 has human tissue and A549 images at 2 μg/ml, while A01574 has corresponding images at 5 μg/ml despite a catalog IF dilution of 4 μg/ml (PB9166 and A01574 IF image captions; A01574 catalog dilution). For IHC across human, mouse and rat, choose A01574 because its IHC images cover all three species; M01574 is a rabbit monoclonal option limited to human reactivity, with a human-colon IHC image (A01574 IHC image captions; M01574 catalog applications and reactivity; M01574 IHC image caption).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry P51636 (CAV2_HUMAN, Caveolin-2).
  2. Human Protein Atlas. CAV2 tissue IHC expression (reliability: Enhanced).
  3. Human Protein Atlas. CAV2 subcellular location (ICC-IF): Localized to vesicles..
  4. Human Protein Atlas. CAV2 antibody validation summary (2 antibodies).
  5. Caveolin 1 and Caveolin 2 are associated with breast cancer basal-like and triple-negative immunophenotype. British journal of cancer 2008 — PMC2480981.
  6. P/Q Type Calcium Channel Cav2.1 Defines a Unique Subset of Glomeruli in the Mouse Olfactory Bulb. Frontiers in cellular neuroscience 2018 — PMC6131590.
  7. CAV2 promotes the invasion and metastasis of head and neck squamous cell carcinomas by regulating S100 proteins. Cell death discovery 2022 — PMC9481523.
  8. CAV2 promotes the growth of renal cell carcinoma through the EGFR/PI3K/Akt pathway. OncoTargets and therapy 2018 — PMC6163017.
  9. PubMed PMID:8552590 — UniProt-cited evidence.
  10. PubMed PMID:9361015 — UniProt-cited evidence.
  11. PubMed PMID:10218480 — UniProt-cited evidence.