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- Table of Contents
Plan CAMK2A paraffin IHC around neuronal and neuropil staining in CNS (HPA tissue IHC). Use matched controls when interpreting signal because the tissue evidence cautions that staining may target protein from more than one gene (HPA tissue IHC).
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
| Expected localisation | CNS neuropil; synaptic enrichment expected (HPA tissue IHC; UniProt) | |
| Staining pattern | Neurons and neuropil in CNS; texture unreported (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 |
| Fixation | Keep formalin fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining may include protein from another gene (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported (UniProt) |
The catalog antibody’s IHC protocol (datasheet: M03241) sits alongside published CAMK2A protocols for rabies brain sections (PMC3660221) and human lung cancer microarrays (PMC7264342).
| 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. |
CAMK2A is a protein associated with synapses, postsynaptic densities, dendrites, and dendritic spines (UniProt Q9UQM7: subcellular location). In paraffin sections, expect selective neuronal and neuropil staining in CNS tissue (HPA: tissue IHC profile). CAMK2A has no transmembrane segment (UniProt Q9UQM7: topology). Treat the pattern as supported rather than definitive: HPA reports medium agreement with RNA data and cautions that its antibodies may target proteins from more than one gene (HPA: IHC reliability).
| Neuronal cells and surrounding neuropil stain in cerebral cortex or caudate. | This fits the reported CNS pattern; both sites show high neuronal-cell staining (HPA: tissue IHC). Assess distribution alongside intensity, since an intense deposit alone does not establish specificity (general IHC practice). |
| Neuropil or dendritic regions stain, while nuclei are not the dominant signal. | This is consistent with synaptic and dendritic localization (UniProt Q9UQM7: subcellular location). Predominantly nuclear staining is outside that expected pattern; review controls and antibody specificity before interpreting it as CAMK2A (UniProt Q9UQM7; general IHC practice). |
| Adipocytes, squamous epithelium, or fibroblasts stain strongly. | These selected cell populations were not detected in HPA tissue IHC: adipose adipocytes, esophageal or oral squamous cells, and skin or soft-tissue fibroblasts (HPA: tissue IHC). Consider cross-reactivity or endogenous detection activity; neither is established from the slide alone (general IHC practice). |
| A uniform chromogenic haze covers cells and extracellular spaces. | Diffuse background does not resolve the selective neuronal and neuropil pattern (HPA: tissue IHC profile). Check background in a primary-antibody omission control and review blocking, washes, and detection conditions (general IHC practice). |
| A cerebral cortex positive-control section has no convincing neuronal signal. | HPA reports high staining in cortical neuronal cells (HPA: tissue IHC). First verify section and detection controls, then review the IHC-validated antibody's supplied protocol; one blank section cannot establish absent CAMK2A expression (general IHC practice). |
| Cellular distribution | UniProt places CAMK2A at synapses, postsynaptic densities, dendrites, and dendritic spines (UniProt Q9UQM7: subcellular location). Interpret IHC at the resolution of neuronal cells and neuropil; do not require individual synapses to be resolved in chromogenic sections (general IHC practice). |
| Tissue choice | Cerebral cortex and caudate neuronal cells, and cerebellar granular-layer cells, are reported high (HPA: tissue IHC). Adipocytes and esophageal squamous cells are reported not detected and can provide contrasting cell populations (HPA: tissue IHC). |
| Antibody specificity | HPA assigns Supported IHC status to HPA051783, HPA051785, HPA053973, and CAB004330, while warning that the staining may reflect proteins from more than one gene (HPA: antibody validation; tissue IHC reliability). Match the antibody and controls used before attributing every positive cell to CAMK2A. |
| Isoforms and epitope | UniProt lists two CAMK2A isoforms, A and B, and a protein kinase domain at residues 13–271 (UniProt Q9UQM7: isoforms; domains). The supplied evidence does not locate an antibody epitope, so isoform recognition and any retrieval-specific effect cannot be predicted from these records. |
| IF/ICC Q: Should its pattern set the IHC expectation? | A: Use it only as context. HPA lists plasma membrane and several ciliary or sperm locations as uncertain and cautions that the ICC antibodies may target proteins from multiple genes (HPA: subcellular ICC-IF). The IHC decision should rest on the tissue pattern and its controls (HPA: tissue IHC; general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in cerebral cortex neurons | Possible section, antibody, or detection failure; cortical neurons are reported high (HPA: tissue IHC). | Confirm tissue identity and run the IHC-validated antibody with its supplied IHC-P conditions and an appropriate detection control (general IHC practice). |
| Strong staining appears only in nuclei | A nucleus-dominant pattern does not match the reported synaptic and dendritic locations (UniProt Q9UQM7: subcellular location). | Compare with neuronal and neuropil staining in a positive-control section; review primary-antibody specificity before scoring the nuclear signal (HPA: tissue IHC; general IHC practice). |
| Adipocytes or squamous cells stain strongly | These selected populations are reported not detected (HPA: tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Inspect a primary-antibody omission control, check chromogen and detection background, and compare staining with a CNS positive control (general IHC practice; HPA: tissue IHC). |
| Diffuse brown haze obscures neuropil | Nonspecific background or residual endogenous enzyme activity may obscure a selective pattern (general IHC practice; HPA: tissue IHC profile). | Check the omission control, endogenous-enzyme blocking where applicable, wash quality, and detection exposure before interpreting intensity (general IHC practice). |
| Peripheral glandular staining seems unexpected | High glandular-cell staining is also listed in adrenal gland, endometrium, fallopian tube, and gallbladder (HPA: tissue IHC). | Do not reject the result solely because it is outside CNS; compare its cell distribution with HPA and use antibody-specific controls given the multigene-target caution (HPA: tissue IHC reliability; general IHC practice). |
| IF/ICC signal differs from the paraffin-section result | HPA's listed ICC-IF locations are uncertain, and its ICC summary carries a multigene-target caution (HPA: subcellular ICC-IF). | Interpret the paraffin section against HPA tissue IHC and its own controls; assess IF/ICC separately on its guide page (HPA: tissue IHC; general IHC practice). |
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.
| 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 → |
| 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 → |
Troubleshoot CAMK2A staining in paraffin sections by checking retrieval, compartment, antibody specificity and matched tissue controls before comparing chromogenic signal.
The catalog includes CAMK2A antibodies with IHC images from human tissue and rat brain, and IF images from human MCF-7 cells and rat brain (catalog image captions).
M03241 has IHC images from paraffin-embedded human colon cancer and rat brain; M03241-3 has IHC images from human brain, hepatocellular carcinoma, and mouse brain (catalog image captions). A03241-3 has a paraffin-embedded human breast carcinoma IHC image, A03241T286 has a paraffin-embedded human brain IHC image, and A03241-2 has IF images from MCF-7 cells and paraffin-embedded rat brain (catalog image captions).
Which to pick: For paraffin-section IHC, M03241 offers a rabbit monoclonal with a documented 1:50 IHC example using heat retrieval in EDTA at pH 8.0; the caption does not report the fixative (M03241 catalog entry and IHC caption). For IF/ICC, choose A03241-2 at the listed 5 μg/mL, supported by an MCF-7 cell IF image; it has no listed IHC application (A03241-2 catalog entry and IF caption). For work spanning species, M03241 has human and rat IHC images and listed human, mouse, and rat reactivity; its mouse IHC performance still needs validation in your sample (M03241 catalog entry and IHC captions).