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- Table of Contents
Plan chromogenic SNAP23 IHC in paraffin sections using membranous staining as a tissue reference (HPA tissue IHC). Start with 2–5 μg/ml of A02487-1 (datasheet A02487-1) and compare controls under consistent fixation.
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | Membranous staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Membranous staining; high at fallopian cilia tips (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02487-1) | |
| Positive control | Fallopian tube+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; no transmembrane segment; epitope map unreported (UniProt) |
The catalog antibody’s IHC-P protocol is paired with one published SNAP23 chromogenic IHC method in mouse lung (PMC4613665: Immunohistochemistry).
| Sample | Paraffin-embedded human lymphoma tissue; fixative not specified (datasheet A02487-1) |
| Fixation | Image fixative and duration unreported (datasheet A02487-1); 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 A02487-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02487-1) |
| Primary antibody | Rabbit anti-SNAP23, 2-5 μg/ml (datasheet A02487-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02487-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02487-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SNAP23-positive staining in ciliated cells (tip of cilia) of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Membranous expression in several tissues. No signal in the no-primary control. |
SNAP23 should appear mainly at the cell membrane, with membranous staining in several tissues (UniProt O00161: cell membrane; HPA tissue IHC: membranous expression). It has no transmembrane segment (UniProt O00161 topology). Fallopian tube ciliated cells show high staining at cilia tips; several glandular and hematopoietic cell populations show medium staining (HPA tissue IHC). HPA rates the tissue staining Approved, with medium consistency against RNA expression (HPA tissue IHC).
| Distinct membrane-associated staining in the expected cells, especially strong staining at fallopian tube cilia tips. | This fits the reported pattern. HPA records high staining at cilia tips in fallopian tube ciliated cells and describes membranous expression in several tissues (HPA tissue IHC). Judge the signal in its tissue and cell context. |
| Predominantly nuclear staining, with little discernible membrane signal. | Nuclear dominance conflicts with the reported cell membrane location (UniProt O00161: cell membrane; HPA subcellular: supported plasma membrane). Treat it as a possible artefact and check controls before assigning it to SNAP23. |
| Strong staining in a cell population listed as undetected, such as skeletal muscle myocytes. | This conflicts with that specific HPA observation (HPA tissue IHC: skeletal muscle myocytes not detected). Check cell identification, antibody specificity and endogenous chromogenic activity; a listed negative result does not prove that every specimen must remain negative. |
| Diffuse brown staining across cells and surrounding tissue, without a defined membrane pattern. | The distribution is less consistent with HPA's membranous tissue profile (HPA tissue IHC). Assess background using appropriate controls; diffuse chromogen alone cannot establish which component of the detection workflow caused it (general IHC practice). |
| No staining in the fallopian tube ciliated cells selected as a positive reference. | HPA reports high staining at their cilia tips (HPA tissue IHC). Confirm that ciliated cells are present and review the staining run with controls before interpreting an absent signal as loss of SNAP23. |
| Tissue and cell selection | HPA reports medium staining in adrenal and breast glandular cells and bone marrow hematopoietic cells, alongside high staining at fallopian tube cilia tips (HPA tissue IHC). Compare the same cell population when using a reference section; tissue names alone conceal differences between cells. |
| Negative-reference selection | HPA lists adipocytes in adipose tissue and cardiomyocytes in heart muscle as not detected, while lung macrophages are low (HPA tissue IHC). These are cell-specific observations. A low-staining population is a weaker negative reference than a listed undetected population. |
| Antibody evidence | The HPA tissue profile is Approved but has medium consistency between staining and RNA expression; three listed antibodies have Approved IHC status (HPA tissue IHC; HPA antibodies). Interpret an unexpected pattern with this validation limit in mind, rather than treating a single stained section as confirmation. |
| Molecular form and localization | SNAP23 has two listed isoforms, SNAP-23a and SNAP-23b, and a single annotated chain spanning residues 1–211, with no signal peptide or propeptide (UniProt O00161). Its lack of a transmembrane segment does not contradict its reported cell membrane localization (UniProt O00161 topology and subcellular location). |
| IF/ICC Q&A: What localization is expected? | Mainly plasma membrane staining; HPA also reports primary cilium localization with uncertain support (HPA subcellular). This is a localization cross-check, not an IHC staining instruction. Do not give the uncertain cilium observation the same weight as the supported plasma membrane location. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-reference section has no visible signal. | The selected cells may be absent, or the staining run may have failed; fallopian tube ciliated cells are reported high at cilia tips (HPA tissue IHC). | Confirm the ciliated cells on the section, then review the antibody, retrieval and detection steps against the run controls (general IHC practice). No SNAP23-specific retrieval condition is established by these sources. |
| Only weak staining appears in a chosen reference tissue. | The chosen population may normally be low: HPA lists lung macrophages as low, but fallopian tube ciliated cells as high (HPA tissue IHC). | Check the annotated cell population and compare with a high-staining reference when available (HPA tissue IHC). Do not infer assay failure solely from a low-staining population. |
| A strong nuclear pattern replaces the expected membrane pattern. | That compartment conflicts with the supported plasma membrane location (HPA subcellular) and UniProt's cell membrane annotation (UniProt O00161). The cause cannot be assigned from appearance alone. | Inspect positive and negative controls and reassess antibody specificity and detection background (general IHC practice). Record the compartment as discordant until resolved. |
| Unexpected brown signal appears in a listed undetected cell population. | Possible explanations include cell misidentification, cross-reactivity or endogenous detection activity (general IHC practice); HPA reports the named population as undetected in its sampled tissue (HPA tissue IHC). | Verify cell identity and inspect appropriate negative and endogenous-activity controls (general IHC practice). Compare the exact cell population with HPA, rather than calling the whole tissue positive or negative. |
| The section shows diffuse brown haze that obscures cell boundaries. | Background from the staining or detection workflow is possible (general IHC practice); haze does not match the reported membranous profile (HPA tissue IHC). | Review blocking, washing and chromogen development with run controls (general IHC practice). Score SNAP23 only where a convincing cell-associated pattern can be distinguished from haze. |
| Staining differs between sections prepared under different conditions. | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Check section quality and documented processing differences, then compare sections with the same controls (general IHC practice). Describe the observed difference without attributing it to a SNAP23-specific fixation effect. |
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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.
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | Ciliated cells (tip of cilia) | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Troubleshoot SNAP23 staining in paraffin sections by checking retrieval, membrane localization, controls, and scoring before interpreting DAB signal (datasheet A02487-1; HPA: membranous expression).
Anti-SNAP23 antibodies have IHC figures from human lymphoma, rectal adenocarcinoma and placenta sections (catalog IHC captions). IF data cover T47D cells (A02487-1 IF caption); PA1774 also lists mouse and rat reactivity (catalog).
A02487 has a human rectal adenocarcinoma IHC figure (A02487 IHC caption), while A02487-1 has a human lymphoma IHC figure and T47D cell IF data (A02487-1 IHC and IF captions). PA1774 has a human placenta paraffin-section IHC figure; its catalog lists IHC reactivity with human, mouse and rat (PA1774 IHC caption; catalog applications and reactivity).
Which to pick: For human paraffin-section IHC, choose A02487 or A02487-1; their own captions document EDTA retrieval at pH 8.0 in rectal adenocarcinoma and lymphoma, respectively (A02487 and A02487-1 IHC captions). For IF/ICC, choose A02487-1 because both applications are listed and its IF caption shows T47D cells (A02487-1 catalog applications; IF caption). For mouse or rat IHC, PA1774 lists both species and IHC, although its displayed paraffin figure is human placenta; fixative and clone are unreported for all three antibodies (PA1774 catalog reactivity and applications; PA1774 IHC caption; all three IHC captions and catalog clone fields).