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CAMK2A / Calcium/calmodulin-dependent protein kinase type II subunit alpha · IHC design guide
Design Immunohistochemistry for CAMK2A
Plan CAMK2A chromogenic IHC in paraffin sections using CNS neurons and neuropil as the expected staining pattern (HPA tissue IHC). Compare with a tissue control reported as unstained, and interpret positive staining cautiously because the HPA profile may reflect proteins from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CAMK2A 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 | Neuronal neuropil (HPA tissue IHC); synaptic sites expected (UniProt) | |
| Staining pattern | CNS neurons and neuropil; finer pattern unspecified (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03241) | |
| Positive control | Adrenal gland+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 sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03241) | |
| Caveat | Staining may reflect proteins from more than one gene (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms (A/B); epitope coverage unknown (UniProt) |
Section 1
Recommended CAMK2A IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by one published CAMK2A staining protocol for paraffin-embedded brain sections (PMC3660221).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M03241) |
| Fixation | Image fixative and duration unreported (datasheet M03241); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M03241); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03241) |
| Primary antibody | Rabbit monoclonal (clone AAFF-3) anti-CAMK2A, 1:50 (datasheet M03241) |
| Primary incubation | Overnight at 4 °C (datasheet M03241) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03241) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CAMK2A-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Selective expression in neuropil and neurons in CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet M03241); citrate at pH 6 is a published alternative (PMC3660221).
Section 2
What Is the Expected CAMK2A Staining Pattern?
CAMK2A is associated with synapses, postsynaptic densities, dendritic spines and dendrites, and has no transmembrane segment (UniProt Q9UQM7). In paraffin-section IHC, expect staining in CNS neurons and neuropil (HPA: selective CNS profile). HPA rates its tissue pattern Supported, with medium agreement between staining and RNA data, and cautions that the antibodies may also detect proteins from more than one gene (HPA: tissue IHC).
| Neuronal cells and surrounding neuropil stain strongly in cerebral cortex or caudate; cerebellar granular-layer cells also stain (HPA: High in these sites). | This fits the reported CNS pattern and synaptic association (HPA: tissue IHC; UniProt Q9UQM7). At IHC resolution, neuropil staining cannot establish staining of individual postsynaptic densities; assess the tissue pattern alongside a control for background (general IHC practice). |
| Staining is predominantly nuclear, with little corresponding signal in neurons or neuropil (HPA: selective CNS profile). | A nuclear-only pattern is inconsistent with the listed synaptic, dendritic and postsynaptic locations (UniProt Q9UQM7). Check whether staining persists in the negative detection control and whether counterstain has been mistaken for chromogen (general IHC practice). |
| Adipocytes or skin fibroblasts stain strongly, while the selected neuronal positive control remains weak (HPA: Not detected in those cells; High in CNS neuronal cells). | Consider antibody cross-reactivity or endogenous detection activity (general IHC practice). Cell identity matters: glandular cells in adrenal gland, endometrium and gallbladder also show High staining, so staining outside the CNS alone does not establish an artefact (HPA: tissue IHC). |
| Chromogen spreads across several cell types or extracellular spaces without a clear cellular pattern (general IHC practice). | This is diffuse background rather than a convincing match to the selective CNS profile (HPA: tissue IHC). Review blocking, primary-antibody concentration and wash stringency; endogenous enzyme activity can also contribute with chromogenic detection (general IHC practice). |
| No signal appears in neuronal cells of cerebral cortex or caudate (HPA: High in those cells). | First check section integrity and the assay controls, then review antigen retrieval, primary-antibody dilution and detection steps (general IHC practice). HPA's Supported rating does not guarantee that every antibody or run will reproduce its reported pattern (HPA: tissue IHC). |
💡Expected CAMK2A appearanceCall the IHC result consistent when neuronal cells and neuropil show a clear, comparatively strong signal in CNS positive tissue (HPA: selective CNS profile; High in cortex and caudate); diffuse background or dominant nuclear-only staining is unconvincing (UniProt Q9UQM7; general IHC practice).
| Subcellular context | Synapse, postsynaptic density, dendrite and dendritic spine are listed locations (UniProt Q9UQM7). Use these to interpret a neuronal and neuropil pattern; chromogenic section IHC does not resolve those structures individually (general IHC practice). |
| Tissue and cell context | Cortex and caudate neuronal cells and cerebellar granular-layer cells are High, whereas adipocytes and skin fibroblasts are Not detected (HPA: tissue IHC). The reported High glandular-cell staining means CNS restriction should not be imposed on every section (HPA: tissue IHC). |
| Antibody validation limit | Four listed antibodies have Supported IHC ratings (HPA: HPA051783, HPA051785, HPA053973, CAB004330). HPA also reports medium staining–RNA consistency and possible detection of proteins from more than one gene; a plausible pattern alone is not proof of CAMK2A specificity (HPA: tissue IHC). |
| Molecular features | UniProt lists isoforms A and B, nine modified residues, and no transmembrane segment or signal peptide (UniProt Q9UQM7). The supplied records do not map antibody epitopes or establish how isoforms or phosphorylation alter this IHC signal (UniProt Q9UQM7; HPA: tissue IHC). |
| IF/ICC Q&A: should membrane or cilium staining be expected? | HPA lists plasma membrane and primary-cilium locations as uncertain and warns of antibodies targeting proteins from multiple genes (HPA: subcellular ICC-IF). Treat those ICC-IF observations as provisional; they do not redefine the supported neuronal and neuropil IHC pattern (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| CNS positive tissue has no detectable neuronal signal (HPA: High in cortex and caudate). | The run may have insufficient retrieval or ineffective primary or detection reagents (general IHC practice); target-specific fixation sensitivity is unreported in the supplied records (UniProt Q9UQM7; HPA: tissue IHC). | Verify the positive control and detection controls, then review the chosen retrieval conditions and catalog-antibody dilution against its IHC instructions (general IHC practice). |
| The entire section has diffuse brown staining (general IHC practice). | Excess primary reagent, incomplete washing or endogenous enzyme activity can produce broad chromogenic background (general IHC practice). | Compare a no-primary control, check blocking and endogenous-enzyme quenching, and adjust primary dilution or washes within the validated IHC workflow (general IHC practice). |
| Adipocytes or skin fibroblasts stain as strongly as the CNS control (HPA: Not detected in these cells). | The cell pattern may reflect off-target recognition or endogenous detection activity; HPA flags possible detection of proteins from more than one gene (HPA: tissue IHC; general IHC practice). | Confirm cell identity and review a negative detection control. Compare the neuronal and neuropil pattern with the reported positive tissues before interpreting the signal as CAMK2A (HPA: tissue IHC; general IHC practice). |
| Only nuclei appear positive in an otherwise readable CNS section (HPA: selective neuronal and neuropil profile). | The distribution conflicts with UniProt's synaptic and dendritic locations; counterstain or nonspecific chromogen may be mistaken for target signal (UniProt Q9UQM7; general IHC practice). | Inspect the counterstain and no-primary control, then reassess signal in neuronal cell bodies and neuropil using the same scoring criteria across controls (general IHC practice). |
| A glandular section is positive despite an expectation of CNS-only staining (HPA: tissue IHC). | HPA reports High glandular-cell staining in adrenal gland, endometrium, fallopian tube and gallbladder (HPA: tissue IHC). | Record the cell type and compare it with the relevant HPA tissue observation; assess controls and antibody specificity before calling the glandular signal false (HPA: tissue IHC; general IHC practice). |
| A low-level tissue appears negative beside a strong CNS control (HPA: tissue IHC). | HPA lists only Low staining in parathyroid glandular cells, colon peripheral nerve or ganglion, and smooth muscle cells; weak signal can be harder to score (HPA: tissue IHC; general IHC practice). | Score the weak tissue separately from the CNS positive control and use matched negative controls to define background; avoid treating absence of visible weak signal alone as assay failure (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CAMK2A staining
M03241 · IHCIHC analysis of CAMK2A using anti-CAMK2A antibody (M03241). CAMK2A was detected in a paraffin-embedded section of human colon cancer tissue.
M03241 · IHCIHC analysis of CAMK2A using anti-CAMK2A antibody (M03241). CAMK2A was detected in a paraffin-embedded section of rat brain tissue. Heat med
M03241-3 · IHCHuman brain was stained with anti-CaMKII α rabbit antibody
M03241-3 · IHCHuman hepatocellular carcinoma was stained with anti-CaMKII α rabbit antibody
A03241-3 · IHCImmunohistochemistry (IHC) analyzes of CaMKIIα/δ (M281) pAb in paraffin-embedded human breast carcinoma tissue at 1:100.
A03241T286 · IHCImmunohistochemistry (IHC) analyzes of p-CaMKIIα/δ (T286) pAb in paraffin-embedded human brain tissue.
A03241-2 · IFIF analysis of CaMKII alpha/CAMK2A using anti-CaMKII alpha/CAMK2A antibody (A03241-2). CaMKII alpha/CAMK2A was detected in immunocytochemica
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000070808-CAMK2A/tissue/cerebral+cortex#imid_14805140
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000070808-CAMK2A/tissue/colon#imid_14805074
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000070808-CAMK2A/tissue/liver#imid_14805075
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000070808-CAMK2A/tissue/kidney#imid_14805076
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA051785; green = CAMK2A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA051785; green = CAMK2A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CAMK2A IHC & IF
🧪Run cerebral cortex first: neuronal cells should stain at the High HPA level (HPA: cerebral cortex neuronal cells, High). Run adipose tissue as the negative comparator because adipocytes are Not detected (HPA: adipose tissue adipocytes, Not detected); on the cortex slide, use non-neuronal cells without specific staining as an internal background comparator, without assuming every such cell is negative (standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CAMK2A in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, with annotated localisation: Plasma membrane (uncertain), Primary cilium tip (uncertain), Primary cilium transition zone (uncertain), Principal piece (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a matched rabbit IgG isotype control, matched for clonality if known; and CAMK2A knockout tissue or a validated peptide-block control (selected-SKU caption: rabbit primary; standard IHC practice). Block endogenous peroxidase for chromogenic detection, and assess tissue autofluorescence if using IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU tissue-IHC caption). The caption uses heat-mediated retrieval in EDTA, pH 8.0, for human colon cancer tissue; whether cerebral cortex staining depends on that retrieval condition is unreported (selected-SKU tissue-IHC caption). The supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC; for cortical IF, check unstained tissue for autofluorescence before interpreting signal (standard IF practice).
HPA tissue IHC evidence for CAMK2A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CAMK2A IHC Tips
Use the selected paraffin-section IHC image as a starting point, then assess staining against neuronal distribution and appropriate controls (datasheet M03241; HPA tissue IHC).
How should I optimize retrieval when CAMK2A staining is weak?
Start paraffin-section chromogenic IHC with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M03241). The selected image used this retrieval before a 1:50 primary incubation overnight at 4°C and peroxidase detection with DAB (datasheet M03241). If staining is weak, compare heating durations on adjacent sections while keeping the buffer and detection conditions consistent (standard IHC practice). Include cerebral-cortex neuronal cells as a positive reference, and inspect tissue morphology for damage from excessive heating (HPA tissue IHC: High in cerebral-cortex neuronal cells; standard IHC practice). Record the retrieval duration and score signal against background under identical imaging conditions (standard IHC practice).
Can I assume the catalog image establishes an optimal fixative?
No fixative is reported for the selected paraffin-embedded human colon cancer section, so target-specific fixation sensitivity remains unknown (datasheet M03241). Do not infer a preferred fixative or fixation duration from its EDTA pH 8.0 retrieval and 1:50 antibody incubation (datasheet M03241). For a new specimen series, document the actual fixation conditions and compare matched sections processed with the same retrieval and detection steps (standard IHC practice). Include a positive neuronal reference because CAMK2A staining is reported in cerebral-cortex neuronal cells (HPA tissue IHC: High in cerebral-cortex neuronal cells). Interpret differences only after checking tissue preservation and background across the matched sections (standard IHC practice).
Which staining pattern fits CAMK2A in paraffin sections?
Prioritize neuronal and neuropil staining, particularly dendritic and synaptic regions, when reviewing chromogenic sections (HPA tissue IHC: selective expression in CNS neuropil and neurons; UniProt Q9UQM7: dendrite, dendritic spine and postsynaptic density). CAMK2A has no transmembrane segment, so an exclusively sharp cell-surface outline warrants scrutiny rather than automatic acceptance (UniProt Q9UQM7: topology; standard IHC practice). Compare the pattern with nearby neuronal cells and tissue architecture on the same section (standard IHC practice). Report compartment and cell type separately, since diffuse tissue color can obscure a localized neuronal pattern (standard IHC practice). Treat reported ciliary and membrane signals cautiously because those antibody observations may include proteins from multiple genes (HPA subcellular: uncertain locations and multi-gene caution).
Does this stain distinguish CAMK2A isoforms or phosphorylation states?
The supplied record lists CAMK2A isoforms A and B, but the selected IHC caption does not identify the antibody epitope or establish isoform selectivity (UniProt Q9UQM7: isoforms; datasheet M03241). It also does not establish recognition of phosphorylated Thr-286, an autophosphorylation site (UniProt Q9UQM7: modified residues; datasheet M03241). Ask for epitope information and test an independently validated reagent before assigning staining to one isoform or activation state (standard IHC practice). Interpret apparent staining alongside controls for related kinase chains, since CAMK2A can assemble with CAMK2B, CAMK2G and CAMK2D (UniProt Q9UQM7: subunit). Tissue staining alone cannot resolve those molecular distinctions (standard IHC practice).
How can I adapt the IHC readout to multiplex IF?
Use the chromogenic IHC pattern to select regions, then establish the IF staining conditions separately; the selected caption documents paraffin-section IHC with DAB (datasheet M03241). Pair CAMK2A with an independently validated neuronal marker to assess the expected cellular pattern (HPA tissue IHC: selective expression in CNS neurons and neuropil; standard IF practice). Choose spectrally separated fluorophores and favor a longer-wavelength CAMK2A channel when tissue autofluorescence interferes (standard IF practice). CAMK2A has no transmembrane segment and is associated with dendrites and postsynaptic densities, so optimize permeabilisation for intracellular epitope access while preserving structure (UniProt Q9UQM7: topology and subcellular location; standard IF practice). Include single-color and secondary-only controls to assess bleed-through and background (standard IF practice).
What should I check when DAB appears across the section?
Check a section without primary antibody to separate detection-system background from primary-associated staining (standard IHC practice). The selected workflow used 10% goat-serum blocking, a 1:50 primary overnight at 4°C, peroxidase-linked detection and DAB (datasheet M03241). If background remains high, reassess serum blocking, primary concentration, washes and the general peroxidase-block step on matched sections (standard IHC practice). Examine edges, damaged regions and pigment before scoring, since these can resemble diffuse chromogen deposition (standard IHC practice). Compare with expected neuronal and neuropil staining and a negative reference, such as adipocyte-rich tissue, rather than relying on overall brown intensity (HPA tissue IHC: CNS profile and adipocytes Not detected; standard IHC practice).
How should I score CAMK2A staining across cases? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and region before scoring, because neuronal and neuropil signals need different denominators (HPA tissue IHC: selective expression in CNS neurons and neuropil; standard IHC practice). For cellular staining, report percent positive neuronal cells and a prespecified intensity-weighted H-score; normalize to the number of evaluable neuronal cells (standard IHC practice). For neuropil, quantify positive area or optical density per mm² of evaluable neuropil, using identical DAB development and image settings (standard IHC practice). Exclude folds, necrosis and edges with a documented rule, and score a matched positive reference in each batch (standard IHC practice). Record retrieval and primary incubation conditions, including EDTA pH 8.0, because processing differences complicate comparisons (datasheet M03241; standard IHC practice).
How do I distinguish convincing CAMK2A signal from artefact?
A convincing chromogenic result should follow identifiable neuronal or neuropil structures, consistent with the reported CNS pattern and dendritic/postsynaptic location (HPA tissue IHC: selective expression in CNS neurons and neuropil; UniProt Q9UQM7: subcellular location). Question isolated edge staining, necrotic patches, diffuse nuclear color or staining that persists without primary antibody (standard IHC practice). Compare the section with a cerebral-cortex neuronal positive reference and a negative reference such as skin fibroblasts (HPA tissue IHC: High in cerebral-cortex neuronal cells; skin fibroblasts Not detected). Check peroxidase blocking and tissue pigment before treating DAB deposits as target signal (standard IHC practice). Avoid claiming isoform or phosphorylation specificity from morphology alone, because those properties are not established by the selected caption (datasheet M03241; standard IHC practice).
Boster reagents
Best CAMK2A / Calcium/calmodulin-dependent protein kinase type II subunit alpha IHC Antibodies
Anti-CAMK2A antibodies have IHC images from human and rat paraffin sections and an IF image from human cells (catalog image captions); all five list human, mouse and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-CaMKII alpha CAMK2A Rabbit Monoclonal Antibody
Cat # M03241
Real IHC data
Anti-CaMKII α Rabbit Monoclonal Antibody
Cat # M03241-3
Real IHC data
Anti-CaMKII Alpha / Delta (M281) CAMK2A Antibody
Cat # A03241-3
Real IHC data
Anti-CaMKII Alpha / Delta (phospho-T286) CAMK2A Antibody
Cat # A03241T286
Real IF data
Anti-CaMKII alpha/CAMK2A Antibody ®
Cat # A03241-2
M03241 has IHC images from human colon cancer and rat brain; M03241-3 from human and mouse brain and human hepatocellular carcinoma (catalog IHC image captions). A03241-3 has an IHC image from human breast carcinoma, A03241T286 from human brain, and A03241-2 an IF/ICC image from MCF-7 cells (catalog image captions).
Which to pick: For paraffin-section IHC, choose rabbit monoclonal M03241: its images document EDTA retrieval at pH 8.0, 1:50 primary antibody and DAB detection; the fixative is unreported (catalog IHC image captions; catalog clone). For IF/ICC, choose A03241-2 at 5 μg/mL for its MCF-7 cell image and listed IF/ICC applications (catalog IF image caption; catalog applications). For cross-species tissue IHC, consider rabbit monoclonal M03241-3: human, mouse and rat reactivity is listed, while its IHC images show human and mouse tissues; the captions do not report fixation or retrieval (catalog reactivity; catalog IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UQM7 (KCC2A_HUMAN, Calcium/calmodulin-dependent protein kinase type II subunit alpha).
- Human Protein Atlas. CAMK2A tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CAMK2A subcellular location (ICC-IF): Mainly localized to the plasma membrane, primary cilium tip, primary cilium transition zone and principal piece. In addition localized to the cell junctions, perinuclear theca, calyx, connecting piece and end piece. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. CAMK2A antibody validation summary (4 antibodies).
- Quantitative proteomics for identifying biomarkers for Rabies. Clinical proteomics 2013 — PMC3660221.
- CAMK2A supported tumor initiating cells of lung adenocarcinoma by upregulating SOX2 through EZH2 phosphorylation. Cell death & disease 2020 — PMC7264342.
- NF2 can mediate the expression of CAMK2A in a tissue specific manner. Scientific reports 2024 — PMC11685432.
- APP Overexpression Causes Aβ-Independent Neuronal Death through Intrinsic Apoptosis Pathway. eNeuro 2016 — PMC4967816.
- PubMed PMID:10231032 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
- PubMed PMID:11972023 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CAMK2A (Calcium/calmodulin-dependent protein kinase type II subunit alpha). Expected localisation Neuronal neuropil (HPA tissue IHC); synaptic sites expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03241). Validated IHC antibody: M03241 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M03241); 2. Fixation: Image fixative and duration unreported (datasheet M03241); 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 retrieval: EDTA pH 8.0 (datasheet M03241); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03241); 8. Primary antibody: Rabbit monoclonal (clone AAFF-3) anti-CAMK2A, 1:50 (datasheet M03241); 9. Primary incubation: Overnight at 4 °C (datasheet M03241). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03241); Caveat: Staining may reflect proteins from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms (A/B); epitope coverage unknown (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q9UQM7, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane, primary cilium tip, primary…, Datasheet M03241, PMC3660221. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CAMK2A/immunohistochemistry-ihc-guide.