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CAMKV / CaM kinase-like vesicle-associated protein · IHC design guide
Design Immunohistochemistry for CAMKV
Plan chromogenic CAMKV IHC in paraffin sections using human brain staining as a reference (datasheet: A30525). Start with the catalog antibody’s 1:100–1:300 IHC range (datasheet: A30525), then assess glia, cortical neuropil and cerebellar granular cells separately (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CAMKV 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 | Cortical neuropil; cerebellar granular-cell cytoplasm/membrane (HPA tissue IHC) | |
| Staining pattern | Medium CNS staining in glia, neuropil and granular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Caudate+3 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining varies by CNS region and cell type (HPA tissue IHC) | |
| Regulation | No intensity regulator annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope impact is unknown (UniProt) |
Section 1
Recommended CAMKV IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published CAMKV tumor pathology protocol (PMC11186719).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A30525) |
| Fixation | Image fixative and duration unreported (datasheet A30525); 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-CAMKV, 1:100 - 1:300 (datasheet A30525) |
| 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 | CAMKV-positive staining in glial cells of caudate (HPA tissue IHC: Medium). HPA tissue profile: Selective expression in CNS. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval at 95–98 °C for 20 min (page retrieval rule). Use high-pH retrieval to reproduce the published protocol (PMC11186719).
Section 2
What Is the Expected CAMKV Staining Pattern?
For chromogenic IHC of paraffin sections, expect selective CNS staining of cerebellar granular cells, cortical neuropil, and glial cells in caudate and hippocampus, reported at medium levels (HPA tissue IHC: Enhanced). Cytoplasmic and membrane associated signal fits CAMKV localisation at the soma plasma membrane and in neurites; UniProt lists no transmembrane segment (UniProt Q8NCB2: subcellular location and topology).
| Medium cytoplasmic or membrane associated staining in cerebellar granular cells, with signal in cortical neuropil (HPA tissue IHC). | This matches reported positive sites and is compatible with CAMKV association with soma membrane and neurites (HPA tissue IHC; UniProt Q8NCB2: subcellular location). Interpret each structure separately: neuropil staining does not identify an individual cell type (standard IHC interpretation). |
| Predominantly nuclear chromogen in the IHC section, without the expected CNS pattern (HPA tissue IHC). | Treat this as an unexpected IHC result and check specificity and scoring (HPA tissue IHC; standard IHC practice). HPA separately reports approved nucleoplasmic staining in ICC-IF, so nuclear signal across the two assays cannot be dismissed or equated without assay specific validation (HPA subcellular ICC-IF). |
| Strong staining in appendix glandular cells or bronchus respiratory epithelial cells (HPA tissue IHC: not detected). | Those cells are reported as negative in HPA tissue IHC. Check for cross-reactivity and, with chromogenic detection, endogenous enzyme activity before attributing the signal to CAMKV (HPA tissue IHC; standard IHC practice). A negative tissue result alone does not establish the cause. |
| Diffuse chromogen across tissue and background, with cell boundaries difficult to distinguish (standard IHC interpretation). | An uninterpretable distribution cannot support the selective CNS pattern reported by HPA. Review blocking, antibody concentration, washes, and detection controls as general IHC variables; no CAMKV specific background mechanism is supplied (HPA tissue IHC; standard IHC practice). |
| No visible staining in cerebellar granular cells or cortical neuropil (HPA tissue IHC: medium). | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
💡Expected CAMKV appearanceCall IHC positive when medium cytoplasmic or membrane associated signal appears in cerebellar granular cells, cortical neuropil, or reported CNS glia; isolated strong staining in HPA-negative epithelial cells is suspect (HPA tissue IHC; UniProt Q8NCB2: subcellular location).
| Tissue and cell context | HPA reports selective CNS expression, with medium IHC staining in specified brain cells and neuropil; several sampled non-CNS cell types are not detected (HPA tissue IHC). Use the named cells when comparing positive and negative regions, rather than treating every brain cell as positive. |
| Subcellular association and topology | UniProt places CAMKV at the cell membrane and cytoplasmic vesicle membrane, predominantly near soma plasma membrane and neurites, while reporting no transmembrane segment (UniProt Q8NCB2: subcellular location and topology). These annotations guide interpretation but do not guarantee a crisp membrane outline in paraffin sections. |
| Isoforms and modified residues | UniProt lists 3 isoforms and phosphothreonine at residues 435 and 459 (UniProt Q8NCB2: isoforms and modified residues). Epitope location and phospho sensitivity are unspecified here; do not infer which isoforms or states an antibody detects. |
| Assay specific localisation evidence | HPA tissue IHC reports granular cell cytoplasm/membrane, neuropil, and glial staining, while HPA ICC-IF reports approved nucleoplasmic and cytokinetic bridge localisation (HPA tissue IHC; HPA subcellular ICC-IF). Keep those observations tied to their respective assays when scoring an IHC slide. |
| IHC evidence and preparation limits | HPA rates the tissue IHC profile Enhanced and lists 2 antibodies with Enhanced IHC validation, HPA007656 and CAB034931 (HPA tissue IHC; HPA antibodies). Neither that rating nor UniProt topology establishes CAMKV specific retrieval conditions or fixation sensitivity; validate preparation choices experimentally (standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cerebellum or cortex has no CAMKV chromogen (HPA tissue IHC: medium). | The tissue region may be misidentified, or a general retrieval, antibody, or detection step may have failed (standard IHC practice); no CAMKV specific failure mechanism is documented here. | Confirm the annotated granular cell or neuropil region, inspect positive and detection controls, and review the validated IHC protocol and retrieval settings (HPA tissue IHC; standard IHC practice). |
| Nuclear staining dominates a paraffin brain section (HPA tissue IHC: expected CNS pattern). | It departs from the reported tissue IHC distribution, although HPA independently records approved nuclear localisation in ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). | Score the IHC compartment separately, check controls, and confirm the pattern with independent antibody evidence where available; do not use ICC-IF localisation alone to validate the IHC result (HPA antibodies; standard IHC practice). |
| Appendix glandular cells or bronchus epithelium stain strongly (HPA tissue IHC: not detected). | Specificity or chromogenic endogenous activity may contribute; the appearance alone cannot distinguish them (HPA tissue IHC; standard IHC practice). | Compare a known-positive CNS region and a detection-only control, then evaluate blocking and antibody specificity under the same IHC conditions (HPA tissue IHC; standard IHC practice). |
| Background is widespread and obscures cell borders (standard IHC interpretation). | Blocking, antibody concentration, washing, or detection chemistry can raise general IHC background; this payload supplies no CAMKV specific cause (standard IHC practice). | Review reagent controls, blocking, washes, and the antibody's validated IHC dilution before scoring localisation (standard IHC practice). |
| Only some expected CNS structures stain (HPA tissue IHC: reported sites are cell specific). | Caudate and hippocampus entries name glial cells, cerebellum names granular cells, and cortex names neuropil; these are different structures (HPA tissue IHC). | Check the section's anatomical region and score each reported cell or structure separately; avoid calling all unstained neighbouring cells assay failures (HPA tissue IHC; standard IHC interpretation). |
| Q: Should the HPA ICC-IF nuclear pattern replace the IHC-P expectation? | HPA ICC-IF reports approved nucleoplasm and cytokinetic bridge localisation, whereas tissue IHC reports selective CNS cell and neuropil staining (HPA subcellular ICC-IF; HPA tissue IHC). | A: No. Interpret the paraffin IHC slide against tissue IHC evidence and its controls; assess ICC-IF on its own guide page and assay evidence (HPA tissue IHC; HPA subcellular ICC-IF; standard IHC interpretation). |
Experiment evidence
IHC & IF images showing CAMKV staining
A30525 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using CAMK5 Antibody. The picture on the right is blocked with the sy
A30525 · IFImmunofluorescence analysis of HeLa cells, using CAMK5 Antibody. The picture on the right is blocked with the synthesized peptide.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164076-CAMKV/tissue/cerebral+cortex#imid_16693119
HPA · HippocampusHuman Protein Atlas IHC, Hippocampus. Source: https://www.proteinatlas.org/ENSG00000164076-CAMKV/tissue/hippocampal+formation#imid_16693120
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164076-CAMKV/tissue/colon#imid_16693053
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164076-CAMKV/tissue/liver#imid_16693046
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA007656; green = CAMKV, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA007656; green = CAMKV, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CAMKV IHC & IF
🧪Run cerebellum first: granular cells should show cytoplasmic or membrane staining at a Medium level (HPA: Cerebellum, granular cells). Run adipose tissue as the negative tissue: adipocytes should show no detectable staining (HPA: Adipose tissue, adipocytes). On the cerebellum slide, cells outside the granular-cell population should remain at background unless independently shown to express CAMKV; HPA does not designate a specific internal negative cell type (HPA: Cerebellum, granular cells).
Positive control tissue: Caudate (Glial cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CAMKV in CACO-2, SH-SY5Y, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched isotype control of the same host species and immunoglobulin class for a monoclonal antibody, or host-matched nonimmune IgG for a polyclonal antibody (standard IHC practice). Use CAMKV knockout material if available or compare with peptide-blocked staining, as illustrated for the catalog antibody in paraffin-embedded human brain tissue (datasheet: A30525 tissue-IHC caption); quench endogenous peroxidase for chromogenic IHC and assess brain autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A CAMKV-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (datasheet: A30525 tissue-IHC caption). Antigen-retrieval dependency and whether frozen sections or IF/ICC are easier are unreported in the supplied evidence; optimize retrieval on paired sections while scoring granular-cell cytoplasmic or membrane staining (HPA: Cerebellum, granular cells). Brain lipofuscin can complicate fluorescence interpretation (standard IF practice), and HPA reports nucleoplasmic and cytokinetic-bridge signal in its ICC-IF images, so assess IF localization separately from tissue IHC (HPA: subcellular).
HPA tissue IHC evidence for CAMKV
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
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Granular cells - cytoplasm/membrane | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuropil | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CAMKV IHC Tips
Troubleshoot CAMKV chromogenic IHC in paraffin sections using the stated retrieval method, CNS staining evidence, and compartment-aware controls.
How should I retrieve CAMKV in paraffin sections when staining is weak?
Use citrate buffer at pH 6.0 for heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule). Keep section thickness, heating, and cooling consistent across the run, then compare a human brain section with a no-primary control before changing retrieval conditions (selected A30525 caption: paraffin-embedded human brain; standard IHC practice). If staining remains weak, test a second retrieval condition on matched sections as a fallback and assess signal alongside tissue damage and background (standard IHC practice). The selected image shows peptide blocking, but its caption reports no retrieval conditions, so it cannot establish a product-specific retrieval optimum (selected A30525 caption).
Can fixation explain weak CAMKV staining in paraffin sections?
CAMKV-specific fixation sensitivity is unknown from the supplied evidence: the selected human brain IHC caption identifies paraffin embedding but does not state the fixative (selected A30525 caption). Record the actual fixative, fixation duration, and processing history for each specimen, then compare matched sections processed with the same pH 6.0 retrieval and detection settings (page retrieval rule; standard IHC practice). Include a documented positive tissue section in each run so a processing problem can be distinguished from a low-signal specimen (HPA: selective CNS expression; standard IHC practice). Do not assign a fixation effect to CAMKV without a controlled comparison using the same antibody and tissue (standard IHC practice).
Which staining compartments are plausible for CAMKV in brain IHC?
Score membrane-associated and neuropil staining with the sampled structure in view: CAMKV is reported at the cell membrane and cytoplasmic vesicle membrane, including the soma membrane and neurites by similarity (UniProt Q8NCB2 subcellular annotation). HPA reports medium staining in cerebral cortex neuropil and cerebellar granular-cell cytoplasm or membrane, while its caudate and hippocampal entries identify glial cells (HPA tissue IHC). HPA ICC/IF instead lists nucleoplasm as its main location and cytokinetic bridge as an additional location, so document antibody, preparation, and compartment when comparing images (HPA subcellular). In chromogenic sections, use morphology and a hematoxylin counterstain to separate cellular signal from dense neuropil and nonspecific precipitate (standard IHC practice).
How can isoforms and epitope placement affect CAMKV IHC?
CAMKV has 3 annotated isoforms, a protein kinase domain at residues 24–286, and phosphothreonines at residues 435 and 459 (UniProt Q8NCB2). Check the catalog antibody's immunogen or mapped epitope against each isoform before interpreting a negative section as absence of all CAMKV forms (UniProt Q8NCB2 isoforms; standard IHC practice). The record lists no transmembrane segment, so do not infer epitope accessibility or membrane orientation from the protein name alone (UniProt Q8NCB2 topology). Compare matched sections with primary omitted and, where appropriate for the selected reagent, peptide competition; the selected human brain image includes a synthesized-peptide block (selected A30525 caption; standard IHC practice).
How should I investigate CAMKV localisation with multiplex IF?
Plan IF as a separate optimisation: the selected catalog image documents paraffin-section chromogenic IHC, while HPA lists ICC/IF images in CACO-2 and SH-SY5Y without supplying fixation or permeabilisation conditions here (selected A30525 caption; HPA subcellular). Pair CAMKV with a validated marker for the cell population being assessed, and check each channel alone before interpreting overlap (standard IF practice). Choose fluorophores and filter sets after measuring tissue autofluorescence in an unstained section, keeping the dimmer signal in a cleaner spectral channel (standard IF practice). Because CAMKV has no annotated transmembrane segment, establish the antibody epitope's accessibility experimentally and titrate permeabilisation only if access to its membrane-associated side requires it (UniProt Q8NCB2 topology; standard IF practice).
What controls help identify nonspecific CAMKV chromogenic staining?
Run a no-primary section and inspect background before assigning brown precipitate to CAMKV; endogenous peroxidase blocking and DAB development are general chromogenic workflow steps (standard IHC practice). Titrate the primary antibody and detection exposure against a brain section, keeping retrieval at citrate pH 6.0, 95–98 °C for 20 min during the comparison (page retrieval rule; standard IHC practice). Inspect section folds, edges, vessels, and damaged areas separately because local staining there can distort an apparent cellular pattern (standard IHC practice). The selected human brain image includes synthesized-peptide blocking, which supports an additional reagent-specific comparison but does not replace no-primary and tissue-context controls (selected A30525 caption; standard IHC practice).
How should CAMKV staining be quantified across brain sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and compartment before scoring, since HPA identifies cerebral cortex neuropil, cerebellar granular-cell cytoplasm or membrane, and glial-cell staining in caudate and hippocampus (HPA tissue IHC). For cellular staining, report percent-positive cells and an H-score using prespecified intensity categories; for neuropil, report positive area or optical density per mm² within a consistent region (standard IHC practice). Normalise cell counts to the relevant cell population or sampled area, and compare optical density only across sections stained and imaged under matched conditions (standard IHC practice). Retain the no-primary baseline and record exclusions for folds or damaged tissue before analysis (standard IHC practice).
When does CAMKV staining support a true tissue signal?
A credible chromogenic result should follow a reproducible CNS pattern and plausible compartment: HPA describes selective CNS expression, while UniProt reports association with soma membrane, neurites, and possible vesicular structures by similarity (HPA tissue IHC; UniProt Q8NCB2 subcellular annotation). Compare the stained cell population with the region-specific HPA observations rather than treating every brown cell as equivalent (HPA tissue IHC). Review unexpected nuclear staining alongside HPA's nucleoplasmic ICC/IF finding, documenting the preparation and antibody before calling it CAMKV in tissue (HPA subcellular; standard IHC practice). Edge accentuation, necrotic areas, and signal surviving no-primary or peroxidase controls warrant artefact review; peptide blocking is available in the selected human brain image as another comparison (selected A30525 caption; standard IHC practice).
Boster reagents
Best CAMKV / CaM kinase-like vesicle-associated protein IHC Antibodies
A30525 has IHC data from paraffin-embedded human brain and IF data from HeLa cells (catalog image captions); reported reactivity covers human, mouse, and rat (catalog: reactivity).
Real IHC data
Anti-CAMK5 CAMKV Antibody
Cat # A30525
A30525 has an IHC image of paraffin-embedded human brain tissue with peptide-blocking control (A30525 IHC image caption). A30525 also has an IF image of HeLa cells with peptide-blocking control and lists IHC, IF, and ICC among its applications (A30525 IF image caption; catalog: applications).
Which to pick: Choose A30525 for paraffin-section IHC based on its human brain image; the fixative is unreported, so the caption does not establish fixation (A30525 IHC image caption). Choose A30525 for IF/ICC because it lists both applications and shows IF in HeLa cells (catalog: applications; A30525 IF image caption). For human, mouse, or rat samples, A30525 is the listed rabbit polyclonal option, though its image evidence is limited to human brain IHC and HeLa IF (catalog: host, clonality, reactivity; A30525 IHC and IF image captions). The selected A30525 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A30525).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NCB2 (CAMKV_HUMAN, CaM kinase-like vesicle-associated protein).
- Human Protein Atlas. CAMKV tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CAMKV subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytokinetic bridge..
- Human Protein Atlas. CAMKV antibody validation summary (2 antibodies).
- Paraneoplastic Calmodulin Kinase-Like Vesicle-Associated Protein (CAMKV) Autoimmune Encephalitis. Annals of neurology 2024 — PMC11186719.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CAMKV (CaM kinase-like vesicle-associated protein). Expected localisation Cortical neuropil; cerebellar granular-cell cytoplasm/membrane (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A30525 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A30525); 2. Fixation: Image fixative and duration unreported (datasheet A30525); 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-CAMKV, 1:100 - 1:300 (datasheet A30525); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining varies by CNS region and cell type (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope impact is unknown (UniProt); Regulation: No intensity regulator annotated (UniProt). Evidence: UniProt Q8NCB2, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A30525, PMC11186719. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CAMKV/immunohistochemistry-ihc-guide.