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CAMKV / CaM kinase-like vesicle-associated protein · IHC design guide
Design Immunohistochemistry for CAMKV
Plan CAMKV IHC in paraffin brain sections using the reported CNS staining patterns as a scoring reference (HPA tissue IHC; datasheet). This guide covers fixation, retrieval, chromogenic detection and antibody titration at 1:100–1:300 (datasheet; standard IHC practice).
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 | Cerebellar granular-cell cytoplasm/membrane (HPA tissue IHC) | |
| Staining pattern | CNS: glia, granular-cell cytoplasm/membrane, neuropil (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 consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | The stained cell type varies by CNS region (HPA tissue IHC) | |
| Regulation | CNS-selective expression (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended CAMKV IHC & IF Protocols
This section pairs the catalog antibody’s IHC-P protocol (datasheet) with a published CAMKV tumor pathology IHC 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 HIER at 95–98 °C for 20 min (page antigen-retrieval rule); compare the published high-pH retrieval for 20 min (PMC11186719).
Section 2
What Is the Expected CAMKV Staining Pattern?
CAMKV is associated with the cell membrane and cytoplasmic vesicle membrane, particularly around the soma and neurites; it has no transmembrane segment (UniProt Q8NCB2, subcellular location and topology). In paraffin IHC, expect selective CNS staining in glial cells, cerebellar granular cells, and cortical neuropil (HPA tissue IHC: Enhanced reliability). Interpret those tissue observations separately from HPA's nuclear ICC-IF pattern.
| Medium staining in caudate or hippocampal glial cells, cerebellar granular-cell cytoplasm/membrane, or cortical neuropil (HPA tissue IHC). | These are the reported positive patterns. Assess the named cells or structures within each section; a uniformly stained brain section would not reproduce HPA's selective CNS profile (HPA tissue IHC). |
| Predominantly nuclear staining in paraffin brain IHC, with little signal in the expected positive cells or neuropil. | This does not match the reported tissue IHC pattern (HPA tissue IHC). Check staining specificity and detection background before assigning CAMKV localisation; HPA reports nucleoplasm in ICC-IF, a separate application (HPA subcellular ICC-IF). |
| Strong staining in adipocytes or colon endothelial cells despite little staining in the expected CNS structures. | Those cell types were reported as not detected (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity (general IHC practice). The HPA result applies to the named cells, not every cell in those tissues. |
| Diffuse chromogen across cells, extracellular spaces, or most of the section, obscuring cellular boundaries. | An uninterpretable diffuse deposit cannot establish CAMKV positivity. Review blocking, antibody concentration, washes, and detection controls (general IHC practice); HPA reports selective CNS expression rather than a diffuse tissue-wide pattern (HPA tissue IHC). |
| No detectable staining in caudate glial cells, cerebellar granular cells, cortical neuropil, and hippocampal glial cells. | All four are reported at medium level (HPA tissue IHC), so first check section quality and the IHC detection workflow with appropriate controls (general IHC practice). One negative field alone does not establish CAMKV absence throughout a brain section. |
💡Expected CAMKV appearanceA convincing positive is medium, selective staining of the specified CNS cells or cortical neuropil (HPA tissue IHC), with membrane-associated localisation plausible (UniProt Q8NCB2); uniform nuclear or broadly diffuse paraffin-IHC staining warrants investigation (HPA tissue IHC; general IHC practice).
| Tissue and cell selection | HPA reports medium signal in four named CNS cell or structure contexts, but no detection in the listed adipocytes and colon endothelial cells (HPA tissue IHC). Compare the specified compartments rather than calling whole tissues positive or negative. |
| IHC antibody evidence | The HPA tissue profile has Enhanced reliability, and HPA007656 and CAB034931 each have Enhanced IHC validation (HPA tissue IHC; HPA antibodies). That supports the reported pattern, while an individual assay still needs its own controls (general IHC practice). |
| Membrane association and processing | CAMKV is associated with cell and cytoplasmic vesicle membranes despite having no transmembrane segment; no signal peptide or propeptide is annotated (UniProt Q8NCB2). These annotations do not predict a specific chromogen distribution in every CNS cell. |
| IF/ICC Q: Is nuclear signal expected? | A: HPA reports approved nucleoplasmic localisation, plus a cytokinetic bridge location, in ICC-IF; images are listed for CACO-2 and SH-SY5Y (HPA subcellular ICC-IF). Evaluate that finding in the separate IF/ICC guide; it does not replace the paraffin-IHC tissue pattern (HPA tissue IHC). |
| Isoforms and epitope coverage | Three isoforms are annotated (UniProt Q8NCB2). The payload gives no antibody epitope or isoform-specific IHC data, so staining cannot be assigned to a particular isoform or assumed to cover all three. |
| Situation | Likely cause | Next action |
|---|---|---|
| No chromogen in an expected positive brain region. | The selected field may lack the HPA-positive cells, or the IHC workflow may have failed (HPA tissue IHC; general IHC practice). | Inspect the specified cells or neuropil and verify section quality, detection controls, and the catalog antibody's IHC-P instructions (HPA tissue IHC; general IHC practice). |
| Signal is weak in an expected positive cell population. | The reported level is medium, so a faint result may be difficult to separate from background (HPA tissue IHC). | Compare a matched positive area with a detection control; review antigen retrieval and antibody dilution using the IHC-validated antibody's instructions (general IHC practice). No CAMKV-specific retrieval condition is supplied. |
| Most of the section shows diffuse brown staining. | Nonspecific binding or detection background can obscure the selective pattern (general IHC practice; HPA tissue IHC). | Review blocking, washes, antibody dilution, and detection controls; score CAMKV only where cells or neuropil can be resolved (general IHC practice). |
| Adipocytes or colon endothelial cells stain strongly. | This conflicts with the reported lack of detection in those cell types (HPA tissue IHC); cross-reactivity or endogenous activity is possible (general IHC practice). | Check a control that omits primary antibody and review the detection system. Reassess the exact cell identity before comparing with HPA (general IHC practice; HPA tissue IHC). |
| Brain IHC shows mainly nuclear staining. | HPA's approved nuclear localisation comes from ICC-IF, whereas the tissue IHC profile identifies glial cells, granular-cell cytoplasm/membrane, and neuropil (HPA subcellular ICC-IF; HPA tissue IHC). | Document the application and morphology, check detection controls, and compare the slide with the reported tissue-IHC pattern before interpreting the nuclear signal (general IHC practice; HPA tissue IHC). |
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 and assess staining in granular cells at the cytoplasm/membrane; HPA rates this signal Medium (HPA: cerebellum, granular cells). Run adipose tissue as a negative comparator because adipocytes are Not detected (HPA: adipose tissue, adipocytes); on the cerebellar slide, use unstained areas as background comparators, without treating other cell types as validated negatives (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 slide and a control immunoglobulin matched to the primary antibody’s host species and isotype or clonality (standard IHC practice). A peptide-blocked section is a product-specific control shown for the catalog antibody in paraffin-embedded human brain tissue (A30525 caption: peptide block); quench endogenous peroxidase for chromogenic IHC, and check brain tissue autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the paraffin-section caption does not name a fixative (A30525 caption: fixative unreported). The evidence does not establish whether frozen sections or IF are easier for CAMKV than paraffin IHC (A30525 caption: paraffin tissue IHC; HPA: ICC-IF images). Brain autofluorescence can complicate IF interpretation, so assess an unstained section alongside any IF experiment (standard IF practice).
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 staining in paraffin sections by checking retrieval, compartment, tissue context, and controls before comparing staining intensity.
What retrieval should I try first for weak CAMKV staining in paraffin sections?
Use citrate pH 6.0 heat-induced retrieval at 95–98 °C for 20 min as the starting condition for CAMKV paraffin IHC (page retrieval rule). Run a human brain section alongside the study samples, because the selected antibody has a paraffin-embedded human brain staining image with peptide blocking (catalog A30525 caption). If staining remains weak, compare a longer exposure to the same citrate buffer on adjacent sections while holding detection conditions constant (standard IHC practice). Check tissue preservation and antibody titration before attributing a negative result to retrieval, since the caption reports neither fixative nor antigen-retrieval conditions (catalog A30525 caption).
Could fixation explain inconsistent CAMKV staining across paraffin blocks?
CAMKV-specific fixation sensitivity is unknown because no target-specific fixation evidence is supplied for this guide (supplied evidence). The selected antibody's brain image establishes staining in paraffin-embedded tissue, but its caption does not identify the fixative or fixation duration (catalog A30525 caption). Compare blocks with documented processing histories, and stain adjacent sections in the same run with the same retrieval and detection settings (standard IHC practice). If signal differs, assess morphology and a known positive control before assigning the difference to CAMKV abundance (standard IHC practice). Do not infer a fixation effect from CAMKV membrane association or phosphorylation sites (UniProt Q8NCB2).
Where should CAMKV staining appear, and how should nuclear signal be handled?
Assess CAMKV chromogen by cell type and compartment: UniProt places it at the cell and cytoplasmic vesicle membranes, particularly around neuronal somata and neurites (UniProt Q8NCB2). Human tissue IHC also reports cortical neuropil and cerebellar granular-cell cytoplasm or membrane staining, while caudate and hippocampal glial cells show medium staining (HPA tissue IHC). HPA cell imaging instead reports mainly nucleoplasmic signal, with additional cytokinetic-bridge localisation (HPA subcellular). Treat isolated nuclear staining in paraffin sections as unresolved until peptide blocking or an independent antibody supports it in that preparation (catalog A30525 caption; standard IHC practice). Compare compartments on well-preserved internal controls before scoring (standard IHC practice).
How can an unknown epitope complicate interpretation of CAMKV IHC?
CAMKV has 3 annotated isoforms, a protein kinase domain spanning residues 24–286, and reported phosphothreonines at 435 and 459 (UniProt Q8NCB2). The supplied caption does not map the catalog antibody's epitope, so its coverage of individual isoforms and sensitivity to those modifications remain unknown (catalog A30525 caption). Request an epitope map before treating unequal staining across samples as an isoform or phosphorylation change (standard IHC practice). Where material permits, compare an independently mapped antibody on adjacent sections and retain the same cell-compartment scoring rules (standard IHC practice). Peptide blocking supports binding specificity in the pictured brain section, without identifying which isoform generated the signal (catalog A30525 caption).
How should I investigate CAMKV patterns with a complementary IF assay?
Use IF as a complementary localisation check for paraffin IHC, with a validated neuronal or glial lineage marker chosen for the structure being scored (HPA tissue IHC; standard IF practice). Place the weaker signal in a red or far-red channel after checking unstained tissue autofluorescence and single-stain controls (standard IF practice). CAMKV lacks an annotated transmembrane segment, and the supplied evidence does not establish which side of an associated membrane contains the antibody epitope (UniProt Q8NCB2; catalog A30525 caption). Test mild permeabilisation when an intracellular epitope is suspected, and compare it with an unpermeabilised control if surface accessibility is plausible (standard IF practice). Confirm any IF pattern against tissue morphology and the chromogenic result (standard IHC/IF practice).
What controls help identify diffuse brown background in CAMKV sections?
First compare a no-primary section with the stained section to identify secondary-detection or tissue-derived chromogen signal (standard IHC practice). Include a peroxidase block before DAB development when using an HRP detection system, and examine whether deposits track vessels, damaged areas, or section edges (standard IHC practice). Titrate the primary antibody and adjust blocking or wash conditions if background persists across compartments (standard IHC practice). The selected brain image includes peptide-blocked tissue, which offers a specificity comparison for that pictured preparation (catalog A30525 caption). Evaluate loss of structured staining separately from loss of diffuse background, because the expected tissue distribution is selective within the CNS (HPA tissue IHC).
How should I quantify CAMKV staining without conflating cell and neuropil signal? ⚠ ANSWER MARKED FOR VERIFICATION
Define regions and compartments before scoring, because reported CAMKV staining includes cortical neuropil, cerebellar granular-cell cytoplasm or membrane, and glial cells in caudate and hippocampus (HPA tissue IHC). For identifiable cells, record the percentage positive and an intensity-based H-score using the same thresholds across slides (standard IHC practice). For neuropil, report positive area or signal density per mm² of viable region, with background subtraction from matched negative controls (standard IHC practice). Normalise cell counts to the number of eligible cells and area measures to the analysed tissue area, excluding folds, edges, and necrosis by a preset rule (standard IHC practice). Keep compartments separate in the final report (standard IHC practice).
When is a CAMKV-positive result credible rather than an IHC artefact?
A credible result has reproducible staining in an appropriate compartment and tissue context: CAMKV is associated with cell and vesicle membranes, while human IHC shows selective CNS expression (UniProt Q8NCB2; HPA tissue IHC). Check whether the pattern follows neuropil, granular cells, or the reported glial populations instead of only torn edges or necrotic regions (HPA tissue IHC; standard IHC practice). Isolated nuclear staining needs separate validation because HPA cell imaging reports nucleoplasmic CAMKV, whereas the tissue and UniProt patterns differ (HPA subcellular; HPA tissue IHC; UniProt Q8NCB2). Use no-primary and peroxidase-block controls to assess endogenous enzyme signal, and compare peptide blocking where feasible (standard IHC practice; catalog A30525 caption).
Boster reagents
Best CAMKV / CaM kinase-like vesicle-associated protein IHC Antibodies
A30525 has IHC data from paraffin-embedded human brain tissue and IF data from HeLa cells; listed reactivity covers human, mouse, and rat (catalog IHC/IF image captions; reactivity).
Real IHC data
Anti-CAMK5 CAMKV Antibody
Cat # A30525
A30525 is listed for IHC and shows staining of paraffin-embedded human brain tissue with a peptide-blocked comparison (catalog applications; IHC image caption). A30525 is also listed for IF/ICC and shows HeLa cell IF with a peptide-blocked comparison; listed reactivity is human, mouse, and rat (catalog applications/reactivity; IF image caption).
Which to pick: For tissue IHC, choose A30525: its own image shows paraffin-embedded human brain tissue, while the fixative is unreported (IHC image caption). For IF/ICC, choose A30525 because both applications are listed and its IF image shows HeLa cells (catalog applications; IF image caption). For cross-species work, A30525 is the listed rabbit polyclonal with human, mouse, and rat reactivity; the pictured IHC sample is human brain tissue (catalog host/clonality/reactivity; IHC image caption).
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 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 consistent across sections (standard IHC practice; not target-specific); Caveat: The stained cell type varies by CNS region (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt; datasheet); Regulation: CNS-selective expression (HPA tissue IHC). 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.