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SV2C / Synaptic vesicle glycoprotein 2C · IHC design guide
Design Immunohistochemistry for SV2C
Plan chromogenic IHC for SV2C in paraffin sections using brain staining as a reference (HPA tissue IHC). Start with the catalog antibody at 1:100 (datasheet: 1:100), then assess cytoplasmic and neuropil staining against the documented tissue pattern (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SV2C 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 | Cytoplasmic in brain and peripheral nerves (HPA tissue IHC) | |
| Staining pattern | Caudate and cerebellar cytoplasm; cortical neuropil (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Cerebellum+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Adrenal nerves stain; glandular cells are undetected (HPA tissue IHC) | |
| Regulation | Condition-dependent regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; cytoplasmic vs extracellular epitope affects access (UniProt) |
Section 1
Recommended SV2C IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by two published SV2C mouse brain IHC protocols (PMC6013333; PMC5073372).
| Sample | Paraffin-embedded Rat Brain tissue; fixative not specified (datasheet A10958) |
| Fixation | Image fixative and duration unreported (datasheet A10958); 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-SV2C, 1:100 (datasheet A10958) |
| 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 | SV2C-positive staining in gLUC cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Distinct cytoplasmic expression in brain, mainly in caudate nucleus, granular layer in cerebellum and in peripheral nerves in pancreas and adrenal gland. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). The published methods used frozen sections (PMC6013333; PMC5073372).
Section 2
What Is the Expected SV2C Staining Pattern?
SV2C is a 12-pass protein of synaptic vesicle membranes, so IHC should favor cytoplasmic or neuropil-associated staining rather than a smooth cell-surface rim (UniProt Q496J9 topology and localization). HPA reports distinct brain staining, especially in caudate nucleus and the cerebellar granular layer, plus peripheral nerves in pancreas and adrenal gland (HPA tissue IHC). Tissue IHC reliability is Supported, with medium agreement between staining and RNA data (HPA tissue IHC).
| Strong cytoplasmic or membrane-associated staining in cerebellar GLUC cells (HPA tissue IHC: High). | This matches an HPA positive tissue and is consistent with a vesicle-associated protein (HPA tissue IHC; UniProt Q496J9 localization). Judge the stained cell population as well as intensity: HPA's High rating applies to the annotated GLUC cells, not every cell in the section (HPA tissue IHC). |
| Moderate staining in cerebral-cortex neuropil, or distinct cytoplasmic staining in caudate (HPA tissue IHC). | Both fit the reported brain distribution: cortical neuropil is rated Medium, and caudate appears in the tissue-profile summary (HPA tissue IHC). Caudate glial cells are separately listed as Not detected; their staining should not be counted as the expected caudate pattern (HPA tissue IHC). |
| Predominantly nuclear staining in paraffin-section IHC, with little expected cytoplasmic or neuropil signal. | Recheck compartment assignment and controls before calling this the expected tissue pattern: UniProt places SV2C on vesicle membranes, while HPA tissue IHC describes cytoplasmic staining (UniProt Q496J9; HPA tissue IHC). HPA separately reports nucleoplasm in ICC-IF, so the discrepancy does not by itself prove an artifact (HPA subcellular ICC-IF). |
| Staining in adrenal glandular cells, adipocytes, or caudate glial cells (HPA tissue IHC: Not detected). | These cell-specific results conflict with HPA's tissue IHC annotations, even where nearby structures may stain: peripheral nerves are reported in adrenal gland (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity, then compare the signal with a no-primary control (general IHC practice). |
| Diffuse chromogen across cells and stroma, or no discernible signal in cerebellar GLUC cells. | Diffuse signal lacks the cell and compartment pattern reported by HPA; check background controls before scoring it as SV2C (HPA tissue IHC; general IHC practice). An absent signal in the annotated High cerebellar cells warrants a run-control check, although HPA rates tissue IHC reliability Supported rather than conclusive (HPA tissue IHC). |
💡Expected SV2C appearanceCall an IHC section positive when cerebellar GLUC cells show High cytoplasmic or membrane-associated staining, with cortical neuropil potentially Medium; diffuse chromogen or staining confined to HPA-negative cell types is suspect (HPA tissue IHC; UniProt Q496J9 localization).
| Tissue and cell selection (HPA tissue IHC) | Use the annotated cerebellar GLUC cells as a positive reference: HPA rates them High, versus Medium cortical neuropil (HPA tissue IHC). Adrenal gland requires cell-level reading because its glandular cells are Not detected although peripheral nerves appear in the tissue-profile summary (HPA tissue IHC). |
| Vesicle topology and unknown antibody epitope (UniProt Q496J9 topology) | SV2C has 12 transmembrane segments and annotated cytoplasmic and extracellular regions (UniProt Q496J9 topology). The supplied record does not identify the antibody epitope; topology alone cannot establish whether a particular antigen-retrieval condition will expose it. |
| Glycosylation and processing (UniProt Q496J9) | UniProt lists 5 glycosylation sites and a full-length chain spanning residues 1–727, with no annotated signal peptide or propeptide (UniProt Q496J9). These annotations do not establish an IHC intensity shift, an epitope-masking effect, or a need for a special retrieval step. |
| Tissue IHC evidence and antibody validation (HPA tissue IHC; HPA antibodies) | HPA calls the tissue pattern Supported and notes medium consistency with RNA, including staining in cells or structures it has not annotated (HPA tissue IHC). Of the listed antibodies, HPA040770 is IHC Supported; HPA040722 has an ICC Approved designation without a listed IHC status (HPA antibodies). |
| IF/ICC Q&A: should nuclear signal be expected in this IHC guide? (HPA subcellular ICC-IF) | HPA ICC-IF reports supported nucleoplasm and approved vesicle localization in SH-SY5Y and U-251MG images (HPA subcellular ICC-IF). That is assay-specific evidence; score paraffin-section IHC against the HPA tissue cytoplasmic and neuropil pattern, and consult the separate IF/ICC guide for that application (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in cerebellar GLUC cells (HPA tissue IHC: High). | The expected positive cell population may have been missed, or the IHC detection run may have failed (HPA tissue IHC; general IHC practice). | Confirm the cell region, inspect a known-positive control from the same run, and verify primary incubation, detection reagents, and counterstain before interpreting a negative sample (general IHC practice). |
| Weak cortical neuropil signal beside strong cerebellar signal. | This difference may reflect HPA's Medium rating for cortical neuropil versus High for cerebellar GLUC cells (HPA tissue IHC). | Score each tissue against its own annotated level and cell compartment; do not require cortex to match cerebellar intensity (HPA tissue IHC). |
| Broad, diffuse chromogen obscures cellular detail. | Nonspecific reagent binding or endogenous detection activity can obscure the expected tissue distribution (general IHC practice; HPA tissue IHC). | Compare with a no-primary control, check blocking and detection steps, and adjust staining conditions using an annotated positive tissue (general IHC practice; HPA tissue IHC). |
| Adrenal glandular cells stain, while nearby nerve profiles are difficult to distinguish. | HPA lists glandular cells as Not detected but reports peripheral nerves in adrenal gland; a section-level positive call could merge those structures (HPA tissue IHC). | Inspect cell morphology and location at higher magnification; use a no-primary control if glandular signal persists (HPA tissue IHC; general IHC practice). |
| Paraffin-section IHC shows mostly nuclear signal. | That differs from HPA's tissue cytoplasmic profile, although HPA separately reports nucleoplasm in ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). | Check positive and no-primary controls, then report the IHC compartment actually observed; do not transfer the ICC-IF localization call to tissue IHC without support (general IHC practice; HPA tissue IHC). |
| A negative result is attributed to formalin sensitivity or failed antigen retrieval. | 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 the positive run control and documented IHC retrieval conditions; if optimizing retrieval, compare conditions on the same annotated positive tissue (general IHC practice; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing SV2C staining
A10958 · IHCImmunohistochemistry (IHC) analysis of paraffin-embedded Rat Brain, antibody was diluted at 1:100.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122012-SV2C/tissue/cerebral+cortex#imid_11049055
HPA · Adrenal glandHuman Protein Atlas IHC, Adrenal gland. Source: https://www.proteinatlas.org/ENSG00000122012-SV2C/tissue/adrenal+gland#imid_11049031
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122012-SV2C/tissue/colon#imid_11090541
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122012-SV2C/tissue/liver#imid_11090534
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA040722; green = SV2C, blue = nucleus, red = microtubules.
HPA · ICC-IF SH-SY5YHuman Protein Atlas immunofluorescence, SH-SY5Y, antibody HPA040722; green = SV2C, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SV2C IHC & IF
🧪Run cerebellum first and look for cytoplasmic or membrane staining in GLUC cells (HPA: High in GLUC cells). Use adipose tissue as the negative tissue, where adipocytes are not detected (HPA: Not detected in adipocytes); other cerebellar cells can serve as internal background comparators if they remain unstained, but HPA does not identify a confirmed negative cell type on that slide.
Positive control tissue: Cerebellum (GLUC cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SV2C in SH-SY5Y, U-251MG, with annotated localisation: Nucleoplasm (supported), Vesicles (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; use SV2C knockout material as a biological specificity control if available (standard IHC practice). Block endogenous peroxidase for chromogenic detection in brain sections, and assess brain autofluorescence before interpreting any IF signal (standard IHC/IF practice).
⚠️Feasibility: The selected A10958 caption shows paraffin-embedded rat brain at 1:100 but does not report the fixative; a target-specific fixation window is unreported (selected IHC caption). An SV2C-specific antigen-retrieval requirement is also unreported, so optimize retrieval with the catalog antibody’s IHC-P protocol (selected IHC caption; standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; brain autofluorescence can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for SV2C
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | 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 → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SV2C IHC Tips
Troubleshoot SV2C chromogenic IHC in paraffin sections using the selected antibody image, tissue staining references, and SV2C topology.
How should I retrieve SV2C in paraffin sections without losing staining specificity?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting). Cool sections consistently before blocking, then compare signal in a brain region with expected cytoplasmic or neuropil staining against a negative tissue on the same run (HPA tissue IHC; standard IHC practice). If staining remains weak, vary retrieval time cautiously while holding the buffer and detection conditions constant; excessive heating can damage morphology and increase background (standard IHC practice). The selected antibody image documents paraffin-embedded rat brain at 1:100, but its caption does not report a retrieval method (A10958 caption).
Could fixation explain weak SV2C staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected rat-brain paraffin IHC caption does not state a fixative or fixation duration (A10958 caption). Record the fixative, fixation interval, tissue thickness, and processing schedule for every specimen, then compare sections processed alike before attributing a signal difference to SV2C abundance (standard IHC practice). Use consistent routine formalin fixation and avoid comparing poorly preserved areas with intact regions, because variable tissue preservation can alter chromogenic staining and morphology (standard IHC practice). Do not infer a fixation effect from SV2C’s membrane topology, phosphorylation sites, or HPA tissue pattern; those records do not test fixation sensitivity (UniProt Q496J9 topology and modified residues; HPA tissue IHC).
Where should convincing SV2C staining appear in tissue?
In chromogenic tissue IHC, look first for cytoplasmic or process-associated signal: HPA reports distinct cytoplasmic brain staining, particularly in caudate nucleus and cerebellar granular layer, plus staining in peripheral nerves of pancreas and adrenal gland (HPA tissue IHC). Vesicle-associated staining fits SV2C’s assignment to synaptic-vesicle membranes and secretory vesicles (UniProt Q496J9 subcellular location). Interpret each compartment within its cell type: HPA lists medium neuropil staining in cerebral cortex but no detection in caudate glial cells (HPA tissue IHC). HPA also reports nucleoplasmic localisation in cell-based IF, so a nuclear tissue-IHC signal requires independent corroboration before being scored as expected vesicular staining (HPA subcellular; UniProt Q496J9 subcellular location).
How does SV2C topology affect epitope access and antibody interpretation?
SV2C has 12 annotated transmembrane segments and no annotated isoforms in the supplied record, so do not assign a stain to an unreported splice variant (UniProt Q496J9 topology and isoforms). Its amino terminus at residues 1–154 and loop at 342–437 are annotated cytoplasmic, whereas the large loop at 459–578 is annotated extracellular and carries several glycosylation sites (UniProt Q496J9 topology and glycosylation). The selected antibody’s epitope is unspecified here, so its accessibility after paraffin processing cannot be predicted from topology alone (A10958 caption; UniProt Q496J9 topology). If retrieval changes the pattern, check whether expected cytoplasmic or neuropil localisation persists before interpreting stronger DAB as improved specificity (HPA tissue IHC; standard IHC practice).
How can IF help check a disputed SV2C IHC pattern?
Use IF as a separate corroboration experiment and multiplex SV2C with a neuronal marker such as MAP2 to test whether signal belongs to the expected neural cells or processes (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore and include unstained tissue to assess autofluorescence, which can obscure weak brain fluorescence (standard IF practice). Because this antibody’s epitope is unspecified, compare a gently permeabilised condition with an appropriate no-detergent control when testing membrane-sided access; intracellular vesicle epitopes generally require permeabilisation (A10958 caption; UniProt Q496J9 topology; standard IF practice). HPA lists SH-SY5Y and U-251MG cell images and both vesicular and nucleoplasmic IF localisation, but those observations do not validate IF performance for the selected IHC antibody (HPA subcellular; A10958 caption).
What should I check when SV2C DAB staining is diffuse?
Start from the selected rat-brain IHC image’s 1:100 antibody dilution, then titrate the antibody while keeping retrieval, incubation, and detection conditions comparable (A10958 caption; standard IHC practice). Include a no-primary control and a peroxidase block before DAB detection to identify reagent background and endogenous enzyme activity (standard chromogenic IHC practice). Examine whether diffuse colour also appears in adipocytes of adipose tissue, where HPA reports SV2C as not detected, while recognising that a negative reference does not establish antibody specificity by itself (HPA tissue IHC; standard IHC practice). If background persists, check wash stringency, blocking, section drying, and DAB development time against a matched positive section (standard IHC practice).
How should I quantify SV2C when staining is concentrated in neuropil? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and compartment before scoring: HPA describes brain cytoplasmic staining and medium cerebral-cortex neuropil staining, which should not be pooled with unrelated cell types (HPA tissue IHC). For identifiable positive cells, report the percentage positive and an intensity-weighted H-score from 0–300; apply the same thresholds to every section (standard IHC scoring practice). For neuropil, report DAB-positive area or optical density per mm² of annotated neuropil, normalised to analysed tissue area and checked against a matched background control (standard image-analysis practice). Keep section thickness, imaging settings, retrieval, and development consistent, and report counts or area separately for each anatomical region (standard IHC practice).
How do I distinguish genuine SV2C staining from artefact?
Favour staining that follows neural cytoplasm or neuropil in the reported positive brain regions and vesicle-associated localisation, while recording the cell type and compartment rather than calling every brown structure positive (HPA tissue IHC; UniProt Q496J9 subcellular location). Treat strong caudate glial staining cautiously because HPA lists those cells as not detected, despite reporting broader caudate cytoplasmic expression (HPA tissue IHC). Exclude section-edge colour, necrotic areas, and signal reproduced by a no-primary control; these patterns can reflect processing or endogenous peroxidase rather than specific DAB staining (standard chromogenic IHC practice). Nuclear staining deserves separate validation because HPA reports nucleoplasmic IF localisation, whereas the tissue-IHC profile emphasises cytoplasm (HPA subcellular; HPA tissue IHC).
Boster reagents
Best SV2C / Synaptic vesicle glycoprotein 2C IHC Antibodies
A10958 has paraffin-embedded rat brain IHC imagery (catalog IHC image caption). IF is listed without an image (catalog applications; IF image alts). Listed reactivity covers human, mouse, and rat (catalog reactivity).
Real IHC data
Anti-SV2C Antibody
Cat # A10958
A10958 is shown in paraffin-embedded rat brain IHC at 1:100 (catalog IHC image caption). A10958 also lists IF as an application and human, mouse, and rat reactivity, but provides no IF image (catalog applications; catalog reactivity; IF image alts).
Which to pick: Choose A10958 for paraffin-section tissue IHC: its own image shows rat brain at 1:100; the fixative is unreported (catalog IHC image caption). For IF/ICC planning, A10958 lists IF at 1:50, but provides no IF image or ICC validation (catalog applications; catalog IF dilution; IF image alts). For cross-species work, A10958 is a rabbit antibody listed as reactive with human, mouse, and rat; its direct IHC image evidence is limited to rat brain (catalog host; catalog reactivity; catalog IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q496J9 (SV2C_HUMAN, Synaptic vesicle glycoprotein 2C).
- Human Protein Atlas. SV2C tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SV2C subcellular location (ICC-IF): Mainly localized to the nucleoplasm and vesicles..
- Human Protein Atlas. SV2C antibody validation summary (2 antibodies).
- Immunochemical analysis of the expression of SV2C in mouse, macaque and human brain. Brain research 2019 — PMC6013333.
- Synaptic vesicle glycoprotein 2C (SV2C) modulates dopamine release and is disrupted in Parkinson disease. Proceedings of the National Academy of Sciences of the United States of America 2017 — PMC5358362.
- Selective Enhancement of Dopamine Release in the Ventral Pallidum of Methamphetamine-Sensitized Mice. ACS chemical neuroscience 2016 — PMC5073372.
- Synaptic vesicle glycoprotein 2C enhances vesicular storage of dopamine and counters dopaminergic toxicity. bioRxiv : the preprint server for biology 2023 — PMC10326994.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:10470851 — UniProt-cited evidence.
- PubMed PMID:21483489 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SV2C (Synaptic vesicle glycoprotein 2C). Expected localisation Cytoplasmic in brain and peripheral nerves (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10958 at 1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (GLUC cells - cytoplasm/membrane); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Rat Brain tissue; fixative not specified (datasheet A10958); 2. Fixation: Image fixative and duration unreported (datasheet A10958); 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-SV2C, 1:100 (datasheet A10958); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Adrenal nerves stain; glandular cells are undetected (HPA tissue IHC); Isoform / epitope: No isoforms annotated; cytoplasmic vs extracellular epitope affects access (UniProt); Regulation: Condition-dependent regulation not annotated (UniProt). Evidence: UniProt Q496J9, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm and vesicles., Datasheet A10958, PMC6013333, PMC5073372. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SV2C/immunohistochemistry-ihc-guide.