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AKAP12 / A-kinase anchor protein 12 · IHC design guide
Design Immunohistochemistry for AKAP12
Plan AKAP12 paraffin IHC around cytoplasmic staining in seminiferous ducts, endothelium and connective tissue (HPA tissue IHC). This guide highlights high staining in Sertoli cells and Bowman's capsule (HPA tissue IHC) and the catalog antibody's 1:100–1:300 IHC dilution range (datasheet: A02303).
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 tissue staining (HPA tissue IHC) | |
| 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining in unannotated cells may complicate scoring (HPA tissue IHC) | |
| Regulation | Regulation not specified (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unreported (UniProt) |
Section 1
Recommended AKAP12 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with published AKAP12 IHC protocols for colon carcinoma tissue microarrays (PMC11695186) and mouse synovium (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 HIER at 95–98 °C for 20 min (page retrieval); the colon carcinoma study used Tris-EDTA unmasking (PMC11695186).
Section 2
What Is the Expected AKAP12 Staining Pattern?
AKAP12 staining should appear mainly in the cytoplasm, with possible cortical or membrane accentuation: UniProt places AKAP12 at the cell cortex, cytoskeleton and membrane, and reports no transmembrane segment (UniProt Q02952 topology). In tissue IHC, expect staining especially in seminiferous ducts, endothelial cells and connective tissue (HPA tissue IHC). HPA rates the tissue staining reliability Enhanced but reports only medium consistency with RNA data (HPA tissue IHC).
| Cytoplasmic staining in testis Sertoli cells or cells of the kidney Bowman's capsule, with little emphasis on nuclei. | This fits two reported High IHC signals (HPA tissue IHC). Judge the stained cell or structure as well as intensity; a positive area alone does not establish the expected pattern (general IHC interpretation). |
| Cytoplasmic staining in lung endothelial cells, ovary follicle cells or smooth muscle cells. | These are reported Medium signals (HPA tissue IHC). Weaker staining than a High example can therefore be consistent with the reference pattern; compare cells within each section rather than expecting uniform tissue staining (HPA tissue IHC; general IHC interpretation). |
| Predominantly nuclear staining, with no convincing cytoplasmic or cortical signal. | The compartment conflicts with the reported cytoplasmic tissue pattern and cytoplasmic, cortical and membrane localization (HPA tissue IHC; UniProt Q02952 subcellular location). Treat it as suspect until control sections and detection background are checked (general IHC practice). |
| Strong signal in a cell population reported as Not detected, such as bone marrow hematopoietic cells or bronchus respiratory epithelial cells. | That result needs scrutiny for cross-reactivity or endogenous detection activity (HPA tissue IHC; general IHC practice). Interpret the named cell population, not the entire organ: HPA also reports Medium lung endothelial staining (HPA tissue IHC). |
| Diffuse staining across cells and extracellular spaces, or no signal in a known-positive cell population. | Uniform haze is less persuasive than the reported cell-restricted cytoplasmic pattern (HPA tissue IHC). An absent signal in Sertoli cells or the Bowman's capsule is inconclusive until section quality, detection and staining controls are checked (HPA tissue IHC; general IHC practice). |
💡Expected AKAP12 appearanceA convincing positive shows cytoplasmic staining, sometimes accentuated near the cell edge, in the reported cell populations—High in Sertoli cells or the Bowman's capsule and Medium in lung endothelium—while isolated nuclear signal or uniform haze is suspect (HPA tissue IHC; UniProt Q02952 subcellular location; general IHC interpretation).
| Compartment and tissue context | UniProt places AKAP12 in cytoplasm, cell cortex, cytoskeleton and membrane; HPA describes tissue IHC as mainly cytoplasmic (UniProt Q02952 subcellular location; HPA tissue IHC). Score the expected cells and compartment together (general IHC interpretation). |
| Variation across cell populations | HPA reports High staining in Sertoli cells and the Bowman's capsule, Medium in lung endothelium, and Not detected in bone marrow hematopoietic cells (HPA tissue IHC). These are cell-level reference observations, not an expectation that every cell in a tissue shares one intensity (HPA tissue IHC). |
| Antibody evidence and limits | The listed antibodies have Enhanced IHC validation, meaning the pattern was reproduced by independent antibodies or supported by orthogonal data (HPA antibody validation). HPA describes only medium staining–RNA consistency and notes staining in unannotated cells or structures (HPA tissue IHC). |
| Isoforms and epitope uncertainty | UniProt lists 3 AKAP12 isoforms (UniProt Q02952 isoforms). The supplied record gives no antibody epitope or isoform coverage, so a difference between antibodies cannot be assigned to an isoform from these data alone (supplied UniProt and HPA records). |
| Processing and modifications | The supplied UniProt record reports no signal peptide, propeptide, transmembrane segment or glycosylation sites, and lists phosphorylation sites (UniProt Q02952 topology and modifications). It does not establish shedding or a phosphorylation-dependent IHC pattern; neither explains an unexpected slide result without further evidence (supplied UniProt record). |
| IF/ICC Q&A: Where should fluorescence appear? | HPA reports plasma membrane and cytosol localization, both with Enhanced ICC-IF evidence; UniProt also places AKAP12 at the cell cortex (HPA subcellular; UniProt Q02952 subcellular location). This localization helps interpret IF images but does not set an IHC staining intensity (HPA subcellular; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in Sertoli cells or the kidney Bowman's capsule. | Either the assay failed or the sampled positive structure is absent or poorly preserved; the image alone cannot distinguish these possibilities (HPA tissue IHC; general IHC practice). | Confirm the structure on the counterstained section, then check a known-positive control and the detection reagents; assess retrieval using the antibody's validated IHC instructions (HPA tissue IHC; general IHC practice). |
| Most nuclei stain, but expected cytoplasm remains clear. | This differs from HPA's mainly cytoplasmic tissue pattern and UniProt's cytoplasmic, cortical and membrane locations (HPA tissue IHC; UniProt Q02952 subcellular location). | Review counterstain and chromogen appearance, compare a reagent control, and repeat with a validated antibody if the mismatch persists (general IHC practice; HPA antibody validation). |
| Nearly every cell and extracellular area shows diffuse color. | Non-specific background or detection activity can obscure the reported cell-associated pattern (general IHC practice; HPA tissue IHC). | Inspect a no-primary control and the blocking and wash steps; score only resolved cellular staining after background is controlled (general IHC practice). |
| A reported Not detected cell population stains strongly. | Cross-reactivity or endogenous detection activity is possible; HPA's negative designation applies to the named cells, not every structure in that organ (HPA tissue IHC; general IHC practice). | Verify cell identity and compare a no-primary control; if the cell-specific mismatch remains, compare an independently validated antibody (general IHC practice; HPA antibody validation). |
| Endothelial staining varies between tissues. | HPA reports Medium staining in lung endothelial cells but Not detected in colon endothelial cells (HPA tissue IHC). | Record the tissue and cell population before judging a result against its matching HPA reference; do not use one organ's endothelial signal as a universal threshold (HPA tissue IHC). |
| Two antibodies give different AKAP12 patterns. | The supplied record lists 3 isoforms but gives no antibody epitopes or isoform coverage, so the discrepancy has no established molecular explanation here (UniProt Q02952 isoforms; supplied HPA antibody data). | Compare each result with the HPA cell and compartment pattern and include appropriate controls; investigate epitope coverage before attributing the difference to isoforms (HPA tissue IHC; general 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 look for AKAP12 staining in Bowman's capsule (High; HPA: kidney), then use adrenal gland glandular cells as the negative tissue (HPA: Not detected in adrenal gland glandular cells). On the kidney slide, compare Bowman's capsule with adjacent cells that lack staining, but do not assume those cells are confirmed biological negatives (HPA: kidney row identifies Bowman's capsule only).
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) control, an isotype control matched to the primary antibody's host species and clonality, and a biological negative such as knockout tissue or peptide blocking; peptide blocking is shown for the catalog antibody (standard IHC practice; A02303 tissue-IHC caption). For chromogenic kidney IHC, quench endogenous peroxidase and check endogenous biotin background if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window, fixation effect, and antigen-retrieval dependency are unreported in the supplied evidence; the paraffin-section image caption does not state a fixative (A02303 tissue-IHC caption). HPA has ICC-IF images for this target, but the supplied evidence does not establish that IF or frozen sections are easier than paraffin IHC (HPA: subcellular images). In kidney, endogenous biotin can complicate avidin–biotin detection, so check that background when choosing detection chemistry (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 section chromogenic IHC by checking retrieval, cell type, compartment, controls and scoring before interpreting signal.
How should I retrieve AKAP12 when paraffin sections stain weakly?
Start with citrate at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting). If signal remains weak, compare a modestly longer retrieval against the page setting on adjacent sections, while checking whether tissue morphology deteriorates (general IHC practice). Keep section thickness, antibody incubation and chromogen development matched so a change in cytoplasmic signal can be attributed to retrieval (general IHC practice; HPA tissue IHC: cytoplasmic profile). Score endothelial or connective tissue structures separately from neighboring cells because AKAP12 staining varies by cell type (HPA tissue IHC). Record retrieval conditions alongside the antibody and detection settings for each comparison (general IHC practice).
Can I adjust fixation to recover weak AKAP12 staining?
AKAP12-specific sensitivity to fixation is unknown from the supplied evidence; the selected paraffin section caption does not state a fixative (A02303 tissue-IHC caption). First compare sections with documented, consistent processing and inspect morphology before attributing weak staining to fixation (general IHC practice). If archival blocks differ, test them with the same citrate pH 6.0 retrieval at 95–98 °C for 20 min, antibody conditions and development time (page retrieval setting; general IHC practice). Interpret any intensity difference cautiously because processing and cell composition may also differ between blocks (general IHC practice; HPA tissue IHC: cell-dependent profile). Do not infer a fixation effect from localisation alone (UniProt Q02952: localisation).
Which compartment should count as AKAP12 positive in tissue IHC?
Prioritize cytoplasmic staining in anatomically identifiable cells and structures (HPA tissue IHC: cytoplasmic expression mainly in seminiferous ducts, endothelial cells and connective tissue). Peripheral or cortical staining can also fit AKAP12 biology because the protein associates with the cell cortex, cytoskeleton and membrane (UniProt Q02952: subcellular location). It has no annotated transmembrane segment, so a thin border signal should be assessed together with cellular context rather than assumed to identify an integral membrane protein (UniProt Q02952: topology). Compare the distribution with nearby negative cells and a no-primary control, keeping chromogen exposure matched (general IHC practice). Score unequivocal nuclear-only staining separately as unexpected for the supplied localisation evidence (UniProt Q02952: subcellular location).
Could an AKAP12 isoform or epitope explain patchy staining?
AKAP12 has 3 annotated isoforms, so establish which sequence the antibody recognizes before comparing samples or calling a cell negative (UniProt Q02952: isoforms; general IHC practice). The supplied evidence does not map the catalog antibody’s epitope to an isoform, leaving isoform coverage unresolved (supplied antibody evidence). Numerous modified residues are annotated, including phosphoserines at positions 11, 19 and 28, but the evidence does not show that these alter this antibody’s staining (UniProt Q02952: modified residues; supplied antibody evidence). Use adjacent sections and matched processing to distinguish reproducible cellular patterns from uneven antigen exposure (general IHC practice). If using A02303, compare staining with its peptide-blocked image as product-specific supporting evidence (A02303 tissue-IHC caption).
How can IF help check an ambiguous AKAP12 IHC pattern?
Use IF as a complementary localisation check when chromogenic IHC cannot resolve cell boundaries, while keeping the IHC interpretation tied to tissue morphology (general IHC/IF practice). Multiplex AKAP12 with an independently validated endothelial marker when testing endothelial staining, and assess overlap cell by cell (HPA tissue IHC: endothelial expression; general IF practice). Choose spectrally separated fluorophores and inspect unstained tissue channels before assigning faint signal, because tissue autofluorescence can mimic fluorescence (general IF practice). For an intracellular epitope, compare gentle permeabilisation with an unpermeabilised control; membrane association alone does not establish extracellular epitope access because AKAP12 has no transmembrane segment (UniProt Q02952: topology and localisation; general IF practice). Do not transfer IF staining conditions directly into chromogenic IHC scoring (general IHC/IF practice).
How do I separate AKAP12 staining from chromogenic background?
Run a no-primary control through the same secondary reagent and chromogen steps to locate reagent-derived staining (general IHC practice). In enzyme-based detection, check endogenous peroxidase blocking and shorten chromogen development if diffuse color appears across unrelated structures (general IHC practice). Evaluate whether signal follows recognizable cytoplasm in endothelial or connective tissue cells, rather than tissue folds, section edges or debris (HPA tissue IHC: cytoplasmic profile; general IHC practice). Compare a higher-expression reference such as kidney Bowman's capsule with a reported negative compartment such as adrenal glandular cells under matched conditions (HPA tissue IHC: high in Bowman's capsule; not detected in adrenal glandular cells). Record both cellular pattern and control staining before changing antibody concentration (general IHC practice).
What should I measure when AKAP12 staining varies across cell types? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and target cell population before scoring because AKAP12 staining differs among structures within tissues (HPA tissue IHC: cell-dependent profile). For cell-based analysis, report the percentage of positive target cells and an intensity-weighted H-score using 0–3 intensity categories (general IHC scoring practice). For processes or irregular structures, measure positive area or stained structure density per mm² within a predefined region (general image analysis practice). Normalize each result to the eligible target-cell count or measured tissue area, excluding folds, necrosis and missing tissue by the same rule in every section (general IHC scoring practice). Keep retrieval, illumination, threshold and chromogen development consistent across the comparison (general IHC scoring practice).
When is an AKAP12-positive IHC result credible?
A credible result follows the expected cellular compartment and appears in plausible structures, such as cytoplasm in endothelial cells or connective tissue (HPA tissue IHC: cytoplasmic profile). Cortex-associated staining can be compatible with AKAP12, whereas isolated nuclear-only color conflicts with the supplied localisation evidence and needs independent checking (UniProt Q02952: subcellular location; general IHC practice). Compare with a no-primary control and inspect section edges, folds and necrotic areas before treating intense color as specific signal (general IHC practice). In enzyme-based chromogenic detection, consider residual endogenous enzyme activity if color persists without primary antibody (general IHC practice). Interpret a negative result against a matched positive control because expression varies by cell type (HPA tissue IHC: cell-dependent profile).
Boster reagents
Best AKAP12 / A-kinase anchor protein 12 IHC Antibodies
A02303 has IHC data from paraffin-embedded human brain and IF data from HeLa cells (catalog: A02303 image captions). Both antibodies list Human, Mouse, and Rat reactivity (catalog: A02303, A02303-4).
Real IHC data
Anti-AKAP 250 AKAP12 Antibody
Cat # A02303
The sole rendered card, A02303, shows IHC on paraffin-embedded human brain (catalog: A02303 IHC image caption). Its IF image shows HeLa cells (catalog: A02303 IF image caption).
Which to pick: For tissue IHC, choose A02303 because its own image documents paraffin-embedded human brain; the fixative is unreported (catalog: A02303 IHC image caption). For IF, A02303 has a HeLa-cell image; for ICC, A02303-4 lists that application but has no IHC or IF image caption (catalog: A02303 IF image caption; A02303-4 applications and image alts). For cross-species work, both list Human, Mouse, and Rat reactivity and are rabbit polyclonals, so those attributes do not favor either SKU (catalog: A02303, A02303-4 reactivity, host, dilution_raw).
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 tissue staining (HPA tissue IHC). 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 paraffin sections. (standard IHC practice; not target-specific); Caveat: Staining in unannotated cells may complicate scoring (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unreported (UniProt); Regulation: 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.