SNAP23 / Synaptosomal-associated protein 23 · IHC design guide

Design Immunohistochemistry for SNAP23

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.

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for SNAP23 (IHC for SNAP23): expected localisation Membranous staining in several tissues (HPA tissue IHC), antibody A02487-1, validated IHC image, and IHC protocol steps
Printable SNAP23 IHC protocol sheet — expected localisation Membranous staining in several tissues (HPA tissue IHC), antibody A02487-1, controls and protocol steps. Open the full SNAP23 IHC guide →

SNAP23 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 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
Important caveats
Reasons your staining may differ from the expected pattern.
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)
Section 1

Recommended SNAP23 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is paired with one published SNAP23 chromogenic IHC method in mouse lung (PMC4613665: Immunohistochemistry).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human lymphoma tissue; fixative not specified (datasheet A02487-1)
FixationImage fixative and duration unreported (datasheet A02487-1); 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 A02487-1); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet A02487-1)
Primary antibodyRabbit anti-SNAP23, 2-5 μg/ml (datasheet A02487-1)
Primary incubationOvernight at 4 °C (datasheet A02487-1)
DetectionHRP-conjugated secondary, DAB chromogen (datasheet A02487-1)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultSNAP23-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.
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet: A02487-1); compare the published citrate pH 6.0 method for mouse lung (PMC4613665: Immunohistochemistry).
Section 2

What Is the Expected SNAP23 Staining Pattern?

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).

What am I looking at on my slide?
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.
💡Expected SNAP23 appearanceA positive IHC result shows defined membrane-associated staining, with high signal at fallopian tube cilia tips; dominant nuclear staining or diffuse brown haze is suspect (HPA tissue IHC; HPA subcellular: supported plasma membrane).
How each factor affects the staining
Tissue and cell selectionHPA 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 selectionHPA 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 evidenceThe 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 localizationSNAP23 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.
Why is my staining missing, weak or wrong?
SituationLikely causeNext 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.

Sample controls for SNAP23 IHC & IF

🧪Run fallopian tube first and look for staining at the tips of cilia in ciliated cells (HPA: High in fallopian tube ciliated cells). Use heart muscle cardiomyocytes as a negative tissue (HPA: Not detected in cardiomyocytes); on the fallopian tube slide, assess adjacent nonciliated cells for background, without assuming they are SNAP23-negative.
Positive control tissue: Fallopian tube (Ciliated cells (tip of cilia), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SNAP23 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody); use SNAP23-knockout tissue as a biological negative if available. Block endogenous peroxidase and inspect for residual DAB background in the fallopian tube section (selected-SKU caption: HRP/DAB detection; standard chromogenic IHC practice).
⚠️Feasibility: The selected-SKU paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but the fixative is unreported and no target-specific fixation window or retrieval dependency is established (selected-SKU caption). HPA ICC-IF supports plasma-membrane localisation, but the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA: supported plasma-membrane location). Ciliary-tip staining can be spatially limited, so inspect the cilia at sufficient magnification and distinguish signal from surface background (HPA: High at the tip of cilia in fallopian tube ciliated cells).

HPA tissue IHC evidence for SNAP23

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.

Positive expression · recommended positive controls

TissueCell typeLevelEvidenceSource
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 →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
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 →
Section 3

Advanced SNAP23 IHC Tips

Troubleshoot SNAP23 staining in paraffin sections by checking retrieval, membrane localization, controls, and scoring before interpreting DAB signal (datasheet A02487-1; HPA: membranous expression).

Which retrieval conditions should I try first when SNAP23 staining is weak?
Begin with heat-mediated EDTA at pH 8.0 for paraffin-section SNAP23 IHC (datasheet A02487-1). The pictured lymphoma section used this retrieval before 10% goat-serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet A02487-1). Keep section thickness, heating duration, cooling, and detection constant while comparing retrieval exposure across serial sections (standard IHC practice). If staining remains weak, trial another buffer only as a documented fallback, checking tissue integrity and background (standard IHC practice). Include a no-primary control and inspect membrane contrast, since excessive retrieval can obscure compartment-specific staining (standard IHC practice; HPA: membranous expression).
Could fixation explain variable SNAP23 staining between paraffin blocks?
Target-specific SNAP23 fixation sensitivity is unknown here; the pictured paraffin-section lymphoma experiment does not report a fixative (datasheet A02487-1). Record fixative, fixation duration, and processing for each block, then compare matched sections under identical detection conditions (standard IHC practice). Start that comparison with heat-mediated EDTA at pH 8.0 and 2 μg/ml primary overnight at 4°C, the documented staining conditions (datasheet A02487-1). Assess morphology, membrane contrast, and background together before attributing differences to fixation (standard IHC practice; HPA: membranous expression). Membrane localization and phosphorylation establish no particular fixation tolerance (UniProt O00161: cell membrane and modified residues).
What SNAP23 staining pattern should count as plausible in tissue sections?
Prioritize distinct cell-border staining: SNAP23 is assigned to the cell membrane, and tissue IHC shows membranous expression in several tissues (UniProt O00161: subcellular location; HPA: tissue IHC profile). Plasma-membrane localization is supported, while additional primary-cilium localization is uncertain in the subcellular dataset (HPA: subcellular localization). High staining at fallopian-tube cilia tips provides a compartment-specific comparison, whereas staining of lung macrophages is reported as low (HPA: tissue IHC). Compare membrane signal with the counterstain and tissue morphology on matched sections, rather than scoring every brown intracellular deposit as positive (standard IHC practice). Document cell type and compartment separately because their patterns carry different interpretive weight (standard IHC practice).
Can this stain distinguish SNAP23 isoforms or altered epitopes?
Do not assign SNAP-23a versus SNAP-23b from this stain alone: two isoforms are listed, but the antibody epitope is unspecified in the supplied evidence (UniProt O00161: isoforms; datasheet A02487-1: epitope unreported). SNAP23 contains coiled-coil homology regions at residues 14–76 and 146–208, with reported phosphorylation near its amino terminus (UniProt O00161: domains and modified residues). Without epitope mapping, neither isoform recognition nor effects of those modifications on staining can be established (UniProt O00161: isoforms and modified residues; datasheet A02487-1: epitope unreported). Compare serial sections using identical retrieval and detection, then seek an epitope-mapped reagent if isoform specificity matters (standard IHC practice). Report the result as SNAP23 immunoreactivity rather than isoform-specific expression (standard IHC reporting practice).
How should I assess SNAP23 by multiplex IF alongside this IHC result?
Treat IF as a separate assay: the supplied antibody example documents paraffin-section chromogenic IHC, not an IF dilution or fixation condition (datasheet A02487-1). Pair SNAP23 with a marker that identifies the expected cell population, then examine membrane overlap within those cells rather than relying on whole-field fluorescence (standard IF practice; HPA: plasma-membrane localization). Choose a fluorophore away from the tissue’s strongest measured autofluorescence and acquire single-stain and no-primary controls (standard IF practice). Map the antibody epitope before choosing permeabilization: test minimal or no detergent for an accessible surface epitope, and controlled permeabilization for an intracellular-facing epitope (standard IF practice; datasheet A02487-1: epitope unreported). Keep IF intensity and chromogenic IHC scores separate when reporting results (standard assay-reporting practice).
How can I reduce diffuse brown signal without losing SNAP23 membrane staining?
First compare a no-primary slide with the stained section to identify detection-related background, while preserving identical retrieval and DAB development (standard IHC practice). The documented example used 10% goat serum before 2 μg/ml primary overnight at 4°C, followed by peroxidase-linked detection (datasheet A02487-1). Apply an endogenous-peroxidase block and control DAB development as general chromogenic IHC steps, especially when brown deposits appear outside intact cells (standard IHC practice). If the no-primary slide is clean, titrate primary concentration or incubation while checking that cell-border contrast remains visible (standard IHC practice; HPA: membranous expression). Review tissue edges and damaged areas separately from viable cells (standard IHC practice).
How should I score SNAP23 when membrane staining varies across cells? ⚠ ANSWER MARKED FOR VERIFICATION
Define the target cell population and score membrane staining separately from diffuse cytoplasmic or extracellular brown signal (standard IHC practice; HPA: membranous expression). Record the percentage of viable target cells at each intensity and calculate an H-score from 0–300, or report the percentage positive at a prespecified threshold (standard IHC scoring practice). For spatial comparisons, report positive-cell density per mm² of viable tissue or the fraction of positive cells among eligible target cells (standard IHC quantification practice). Keep retrieval, exposure to DAB, counterstain, and scoring thresholds consistent across slides (standard IHC practice). Report compartment and cell type alongside the numerical result, since tissue patterns vary (HPA: tissue IHC profile).
How do I separate true SNAP23 signal from tissue artefacts?
Give greatest weight to reproducible staining at intact cell borders, consistent with SNAP23 membrane assignment and the reported tissue IHC pattern (UniProt O00161: cell membrane; HPA: membranous expression). Treat isolated nuclear staining, necrotic debris, and section-edge deposits cautiously, and compare them with the no-primary control (standard IHC practice). A positive peroxidase-control signal indicates endogenous enzyme or detection background requiring separate investigation before interpreting DAB deposits (standard IHC practice). Compare cell identity as well as location: fallopian-tube cilia tips are reported high, whereas skeletal-muscle myocytes are reported undetected (HPA: tissue IHC). The tissue IHC dataset has approved reliability with medium staining-to-RNA consistency, so corroborate unexpected patterns independently (HPA: reliability).
Boster reagents

Best SNAP23 / Synaptosomal-associated protein 23 IHC Antibodies

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).

Real IHC data IHC analysis of SNAP23 using anti-SNAP23 antibody (A02487-1). SNAP23 was detected in a paraffin-embedded section of human lymphoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SNAP23 Antibody (A02487-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-SNAP23 Antibody ®
Cat # A02487-1
Real IHC data IHC analysis of SNAP23 using anti-SNAP23 antibody (A02487). SNAP23 was detected in a paraffin-embedded section of differentiated adenocarcinoma of the human rectum tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SNAP23 Antibody (A02487) 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-SNAP23 Antibody ®
Cat # A02487
Real IHC data Anti-SNAP23 antibody, PA1774, IHC(P) IHC(P): Human Placenta Tissue
Anti-SNAP23 Antibody ®
Cat # PA1774

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).

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

References

  1. UniProt Consortium. UniProt entry O00161 (SNP23_HUMAN, Synaptosomal-associated protein 23).
  2. Human Protein Atlas. SNAP23 tissue IHC expression (reliability: Approved).
  3. Human Protein Atlas. SNAP23 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the primary cilium..
  4. Human Protein Atlas. SNAP23 antibody validation summary (3 antibodies).
  5. The SNARE protein SNAP23 and the SNARE-interacting protein Munc18c in human skeletal muscle are implicated in insulin resistance/type 2 diabetes. Diabetes 2010 — PMC2911056.
  6. SNAP23 is selectively expressed in airway secretory cells and mediates baseline and stimulated mucin secretion. Bioscience reports 2015 — PMC4613665.
  7. SNAP23 promotes the malignant process of ovarian cancer. Journal of ovarian research 2016 — PMC5114815.
  8. LINC00511 drives invasive behavior in hepatocellular carcinoma by regulating exosome secretion and invadopodia formation. Journal of experimental & clinical cancer research : CR 2021 — PMC8176717.
  9. PubMed PMID:8663154 — UniProt-cited evidence.
  10. PubMed PMID:9070898 — UniProt-cited evidence.
  11. PubMed PMID:11354632 — UniProt-cited evidence.