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AKAP12 / A-kinase anchor protein 12 · IHC design guide
Design Immunohistochemistry for AKAP12
Plan paraffin-section AKAP12 IHC with catalog antibody A02303 at 1:100–1:300 (datasheet). Use the cytoplasmic tissue profile to choose scoring regions, and review unannotated stained structures when interpreting results (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AKAP12 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 (HPA tissue IHC); cortical cytoskeleton possible (UniProt) | |
| Staining pattern | Cytoplasmic in seminiferous ducts, endothelium and connective tissue (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | Adrenal gland+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 | Unannotated stained cells may complicate scoring (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unresolved (UniProt) |
Section 1
Recommended AKAP12 IHC & IF Protocols
The catalog antibody protocol uses citrate retrieval (page antigen-retrieval rule). Two published chromogenic IHC protocols provide additional conditions (PMC11695186; PMC10202715).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A02303) |
| Fixation | Image fixative and duration unreported (datasheet A02303); 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-AKAP12, 1:100-1:300 (datasheet A02303) |
| 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 | AKAP12-positive staining in bowman's capsule of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in seminiferous ducts, endothelial cells and connective tissue. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 at 95–98 °C for 20 min (page antigen-retrieval rule); consider Tris-EDTA for colon carcinoma cores (PMC11695186).
Section 2
What Is the Expected AKAP12 Staining Pattern?
AKAP12 should show predominantly cytoplasmic staining, sometimes accentuated at the cell edge, in selected structures such as testicular Sertoli cells, cells of the kidney Bowman’s capsule, endothelium and connective tissue (HPA: tissue IHC; UniProt Q02952: cell cortex and cytoskeleton). It has no transmembrane segment (UniProt Q02952: topology). HPA rates the tissue IHC evidence “Enhanced” while noting medium consistency with RNA expression (HPA: reliability description).
| Cytoplasmic staining in Sertoli cells or cells of the kidney Bowman’s capsule, with little staining in adjacent cells. | This matches the reported high staining in those specific cell populations (HPA: Testis, Sertoli cells, High; Kidney, Bowman’s capsule, High). Judge the named cells within each tissue rather than treating an entire section as uniformly positive (HPA: tissue IHC). |
| Cytoplasmic staining with some cell-edge accentuation; no dominant nuclear signal. | This fits the reported cytoplasmic IHC profile and AKAP12’s cortical cytoskeleton and membrane associations (HPA: tissue IHC profile; UniProt Q02952: subcellular location). A predominantly nuclear result warrants review of morphology and staining controls; neither source lists the nucleus as an expected main location (HPA: subcellular location; UniProt Q02952: subcellular location). |
| Strong nuclear staining or an exclusively crisp surface outline. | Neither is the expected IHC pattern: HPA describes mainly cytoplasmic tissue staining, and UniProt reports no transmembrane segment (HPA: tissue IHC profile; UniProt Q02952: topology). Check whether counterstain, precipitate or another staining artifact is being mistaken for specific signal (standard IHC practice). |
| Signal dominates HPA-negative cell populations, such as bone marrow hematopoietic cells or adrenal glandular cells. | Those named populations were reported as not detected (HPA: Bone marrow, hematopoietic cells; Adrenal gland, glandular cells). Consider nonspecific antibody binding or endogenous detection activity, and compare primary-antibody omission and detection controls before assigning the signal to AKAP12 (standard IHC practice). |
| Diffuse color across tissue, or no signal in a known-positive cell population. | A widespread haze cannot establish the reported cell-specific pattern (HPA: tissue IHC profile). An absent signal in Sertoli cells or cells of the Bowman’s capsule conflicts with HPA’s high staining reports, but by itself does not identify the failed step (HPA: Testis; Kidney). Review controls and morphology before interpreting the specimen (standard IHC practice). |
💡Expected AKAP12 appearanceCall a result positive when the appropriate cells show predominantly cytoplasmic, possibly cell-edge-accentuated signal—strongest in HPA’s high-staining Sertoli cells or cells of the Bowman’s capsule—while diffuse tissue-wide or dominant nuclear color is suspect (HPA: tissue IHC; UniProt Q02952: subcellular location; standard IHC practice).
| Cell population and tissue | Expected intensity depends on the named cells: Sertoli cells and cells of the Bowman’s capsule are High; lung endothelial, ovarian follicle and smooth muscle cells are Medium (HPA: tissue IHC). Colon endothelial cells are Not detected, so endothelial identity alone does not predict positivity (HPA: Colon). |
| Intracellular distribution | A cytoplasmic pattern can include cortical or membrane-associated emphasis because AKAP12 is reported at the cell cortex, cytoskeleton and membrane (UniProt Q02952: subcellular location). Its lack of a transmembrane segment does not require an uninterrupted surface outline in paraffin IHC (UniProt Q02952: topology). |
| Antibody evidence and isoforms | HPA lists Enhanced IHC status for HPA006344, HPA056230 and CAB026379 (HPA: antibodies). UniProt lists three isoforms, but these records do not establish which isoforms a given antibody detects; check its documented epitope before drawing isoform-specific conclusions (UniProt Q02952: isoforms; standard IHC practice). |
| IF/ICC Q&A: Where should fluorescence appear? | Predominantly in cytosol and at the plasma membrane, both rated enhanced in HPA’s ICC-IF summary (HPA: subcellular). This supports compartment interpretation in IF/ICC; it does not turn the ICC-IF images into a paraffin-IHC protocol or establish matching signal intensity across methods (HPA: subcellular; standard IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in Sertoli cells or cells of the kidney Bowman’s capsule. | The result differs from HPA’s High reports, but the records cannot identify whether tissue quality, detection or antibody performance is responsible (HPA: Testis; Kidney; standard IHC practice). | Confirm the named cells are present, then inspect a concurrent positive control and each detection step. Review the antibody’s documented IHC conditions; do not infer an AKAP12-specific fixation effect from these sources (standard IHC practice). |
| Only nuclei stain strongly. | A dominant nuclear pattern conflicts with the reported cytoplasmic and cortical locations (HPA: tissue IHC profile; UniProt Q02952: subcellular location). | Check the hematoxylin counterstain and chromogen distribution, and compare a primary-antibody omission control. Treat the result as unconfirmed until expected cytoplasmic staining appears in a known-positive cell population (standard IHC practice; HPA: Testis; Kidney). |
| Most cell types show diffuse color. | This obscures the selective cytoplasmic pattern reported by HPA and may reflect nonspecific binding or background detection (HPA: tissue IHC profile; standard IHC practice). | Compare the primary-antibody omission control and a documented negative cell population; inspect blocking, washes, antibody dilution and detection background under the chosen IHC workflow (standard IHC practice; HPA: Bone marrow, hematopoietic cells, Not detected). |
| A nominally negative region stains. | The HPA negative calls apply to specified cells, such as adrenal glandular cells and bone marrow hematopoietic cells, rather than every structure in those tissues (HPA: tissue IHC negatives). | Identify the stained cell type before calling a discrepancy. If the specifically named negative cells stain, compare omission and detection controls and reassess specificity (HPA: tissue IHC negatives; standard IHC practice). |
| Endothelial staining varies between tissues. | HPA reports Medium staining in lung endothelial cells but Not detected in colon endothelial cells (HPA: Lung; Colon). | Score each tissue and cell population against its own HPA observation. Use morphology and appropriate controls when a result differs; do not use lung endothelium as a universal endothelial intensity benchmark (HPA: Lung; Colon; standard IHC practice). |
Experiment evidence
IHC & IF images showing AKAP12 staining
A02303 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using AKAP12 Antibody. The picture on the right is blocked with the s
A02303 · IFImmunofluorescence analysis of HeLa cells, using AKAP12 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/ENSG00000131016-AKAP12/tissue/cerebral+cortex#imid_2000640
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000131016-AKAP12/tissue/colon#imid_2000574
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000131016-AKAP12/tissue/liver#imid_2000583
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000131016-AKAP12/tissue/kidney#imid_2000568
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056230; green = AKAP12, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA056230; green = AKAP12, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AKAP12 IHC & IF
🧪Run kidney first and require staining in Bowman's capsule cells (HPA: High in Bowman's capsule); use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in hematopoietic cells). On the kidney slide, cells lacking specific cortical or cytoplasmic staining can provide an internal background reference, but the supplied HPA rows do not identify a renal cell type that must be negative (UniProt Q02952 subcellular location; HPA: kidney Bowman's capsule High).
Positive control tissue: Kidney (Bowman's capsule, HPA High)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AKAP12 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), HEK293, with annotated localisation: Plasma membrane (enhanced), Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section and an isotype control matched to the primary antibody’s host species, antibody class and concentration; use AKAP12 knockout material or a validated peptide block as a biological specificity control (selected-SKU caption: peptide-blocked image). For kidney chromogenic IHC, quench endogenous peroxidase and check for endogenous biotin if using avidin-biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; the selected A02303 paraffin-section caption does not state the fixative (selected-SKU caption). HPA documents ICC-IF localization at the plasma membrane and in the cytosol, but provides no basis to say frozen sections or IF are easier than paraffin IHC (HPA subcellular: plasma membrane and cytosol). In kidney, assess endogenous biotin as a possible chromogenic detection artefact when using avidin-biotin reagents (standard IHC practice).
HPA tissue IHC evidence for AKAP12
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Kidney | Bowman's capsule | High | Protein (IHC) | HPA → |
| Testis | Sertoli cells | High | Protein (IHC) | HPA → |
| Cerebellum | Processes in molecular layer | Medium | Protein (IHC) | HPA → |
| Lung | Endothelial cells | Medium | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | 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 AKAP12 IHC Tips
Troubleshoot AKAP12 staining in paraffin sections by checking retrieval, compartment, cell identity, and matched controls before interpreting chromogenic signal.
How should I retrieve AKAP12 in paraffin sections when staining is weak?
For AKAP12 IHC on paraffin sections, start with citrate buffer at pH 6.0 for HIER at 95–98 °C for 20 min (page retrieval). Keep section thickness, heating vessel, and cooling conditions consistent across the comparison, then inspect both staining and tissue integrity (standard IHC practice). If staining remains weak, compare a modestly longer retrieval with the prescribed condition on adjacent sections, because excessive heating can damage morphology (standard IHC practice). Score the cytoplasmic or cortical pattern in expected cells rather than treating any increase in diffuse color as improved retrieval (UniProt Q02952 localisation; HPA tissue IHC).
Could fixation explain variable AKAP12 staining between paraffin blocks?
AKAP12-specific sensitivity to any fixative or fixation duration is unknown from the supplied evidence (A02303 caption: fixative not stated). The selected image shows paraffin-embedded human brain tissue, but its caption does not establish how that specimen was fixed (A02303 caption). Compare sections with documented, similar processing histories and run them together using the page’s pH 6.0, 20 min retrieval condition (page retrieval; standard IHC practice). If a poorly staining block also loses morphology or other expected antigens, investigate processing and detection before assigning the difference to AKAP12 biology (standard IHC practice). Do not infer fixation sensitivity from tissue staining patterns or the protein’s topology (HPA tissue IHC; UniProt Q02952 topology).
Should AKAP12 stain the membrane, cytoplasm, or both in IHC?
Expect cytoplasmic staining with possible cortical or membrane-associated accentuation: AKAP12 is reported at the cell cortex, cytoskeleton, and membrane, while HPA reports cytosolic and plasma-membrane localisation (UniProt Q02952 localisation; HPA subcellular). A membrane outline does not imply a transmembrane protein, because AKAP12 has no annotated transmembrane segment (UniProt Q02952 topology). In tissue, inspect endothelial cells, connective tissue, and seminiferous ducts before judging an unfamiliar compartmental pattern (HPA tissue IHC). Compare the same cell type across adjacent sections and record cytoplasmic and peripheral staining separately, since a chromogenic deposit alone cannot resolve a thin cortical compartment reliably (standard IHC practice).
Can isoforms or modified residues explain discordant AKAP12 IHC results?
AKAP12 has 3 annotated isoforms and multiple modified residues, including phosphoserines, so antibody recognition may depend on the epitope actually targeted (UniProt Q02952 isoforms; UniProt Q02952 modified residues). The supplied A02303 caption does not map that epitope or establish isoform coverage (A02303 caption). Its peptide-blocked comparison supports competition by the synthesized peptide in the pictured paraffin-embedded human brain section, but does not by itself establish isoform specificity (A02303 caption; standard IHC interpretation). When sections disagree, compare antibody epitope documentation, specimen processing, and expected cell compartments before attributing the difference to splicing or phosphorylation (UniProt Q02952 isoforms; standard IHC practice).
How can I check AKAP12 localisation by IF alongside chromogenic IHC?
For a separate IF check, multiplex AKAP12 with CD31 to identify endothelial cells, an expected AKAP12-positive cell population in tissue (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore where tissue autofluorescence is troublesome, and inspect unstained and single-label controls before calling colocalisation (standard IF practice). Because the antibody epitope’s membrane-facing side is unreported and AKAP12 has no transmembrane segment, compare staining with and without gentle permeabilisation, such as 0.1% Triton X-100 for 5 min (A02303 caption; UniProt Q02952 topology; standard IF practice). Use IF to assess cytosolic versus peripheral distribution, then evaluate chromogenic IHC on its own matched sections and controls (HPA subcellular; standard IHC/IF practice).
What should I adjust when AKAP12 IHC shows diffuse brown background?
First compare a no-primary control with the test section; color in both points toward detection chemistry or endogenous tissue activity rather than antibody-dependent AKAP12 staining (standard IHC practice). For peroxidase-based DAB detection, include a peroxide block and limit DAB development consistently across sections (standard IHC practice). If background is antibody-dependent, check blocking, wash stringency, and primary concentration without assuming an undocumented AKAP12 dilution (standard IHC practice; A02303 caption). Judge improvement in expected cytoplasmic or cortical staining within annotated cell populations, because uniformly brown stroma or section edges are poor evidence of target localisation (UniProt Q02952 localisation; HPA tissue IHC; standard IHC practice).
How should I quantify AKAP12 IHC across specimens with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since AKAP12 staining is reported in seminiferous ducts, endothelial cells, and connective tissue (HPA tissue IHC). For a consistent population, report percent positive cells and an H-score from 0–300, using intensity categories 0–3 and the percentage in each category (standard IHC practice). For sparse endothelial profiles, positive-cell density per mm² can supplement the score, with the analyzed tissue area documented (standard IHC practice). Normalize comparisons to the number or area of the relevant cells, use matched staining runs and thresholds, and exclude folds, necrosis, and edge artefacts before scoring (standard IHC practice).
How do I distinguish genuine AKAP12 staining from tissue or detection artefact?
A plausible positive signal follows cytoplasmic or cortical distribution in an expected cell population, such as endothelial cells or Sertoli cells (UniProt Q02952 localisation; HPA tissue IHC). Recheck a nuclear-only or uniformly luminal pattern against adjacent morphology and controls, because neither is an annotated dominant AKAP12 location (UniProt Q02952 localisation; HPA subcellular; standard IHC practice). Section-edge accentuation, necrotic regions, and color present in a no-primary control favor processing or endogenous-enzyme artefact (standard IHC practice). The peptide-blocked A02303 image offers a product-specific comparison, but interpret your section using its own matched controls and cell identity (A02303 caption; standard IHC practice).
Boster reagents
Best AKAP12 / A-kinase anchor protein 12 IHC Antibodies
The IHC-validated antibody has paraffin-section human brain data and HeLa IF data (catalog image captions); catalog reactivity covers human, mouse and rat (catalog: A02303).
Real IHC data
Anti-AKAP 250 AKAP12 Antibody
Cat # A02303
A02303 will render with an IHC image of paraffin-embedded human brain tissue and peptide blocking (catalog: A02303 IHC caption). Its IF image shows HeLa cells with peptide blocking, and its listed reactivity is human, mouse and rat (catalog: A02303 IF caption; catalog: A02303 reactivity).
Which to pick: For tissue IHC, choose A02303: its own image documents paraffin-embedded human brain tissue; the fixative is unreported (catalog: A02303 IHC caption). For IF, A02303 has a HeLa image; for ICC, A02303-4 lists ICC and IF but has no supplied image, and both antibodies are rabbit polyclonals (catalog: A02303 IF caption; catalog: A02303-4 applications and image alts; catalog: antibody descriptions and host). For work across human, mouse and rat, either SKU lists all three species, though the supplied IHC image documents human tissue only for A02303 (catalog: both reactivity lists; catalog: A02303 IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q02952 (AKA12_HUMAN, A-kinase anchor protein 12).
- Human Protein Atlas. AKAP12 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. AKAP12 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. AKAP12 antibody validation summary (3 antibodies).
- LSD1 and CoREST2 Potentiate STAT3 Activity to Promote Enteroendocrine Cell Differentiation in Mucinous Colorectal Cancer. Cancer research 2025 — PMC11695186.
- A-kinase anchoring protein 12 is downregulated in human hepatocellular carcinoma and its deficiency in mice aggravates thioacetamide-induced liver injury. Oncology letters 2018 — PMC6176350.
- Loss of AKAP12 aggravates rheumatoid arthritis-like symptoms and cardiac damage in collagen-induced arthritis mice. Experimental animals 2023 — PMC10202715.
- Comprehensive Analysis of the Immune Implication of AKAP12 in Stomach Adenocarcinoma. Computational and mathematical methods in medicine 2022 — PMC9489422.
- PubMed PMID:9000000 — UniProt-cited evidence.
- PubMed PMID:9604001 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AKAP12 (A-kinase anchor protein 12). Expected localisation Cytoplasmic (HPA tissue IHC); cortical cytoskeleton possible (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A02303 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Bowman's capsule); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A02303); 2. Fixation: Image fixative and duration unreported (datasheet A02303); 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-AKAP12, 1:100-1:300 (datasheet A02303); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Unannotated stained cells may complicate scoring (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unresolved (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt Q02952, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane and cytosol., Datasheet A02303, PMC11695186, PMC10202715. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AKAP12/immunohistochemistry-ihc-guide.