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AKAP1 / A-kinase anchor protein 1, mitochondrial · IHC design guide
Design Immunohistochemistry for AKAP1
Plan AKAP1 IHC-P around granular cytoplasmic tissue staining (HPA tissue IHC) and its mitochondrial outer membrane location (UniProt). The catalog antibody has an IHC dilution range of 1:50–1:200 (datasheet); interpret staining alongside the atlas’s medium consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AKAP1 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 | Granular cytoplasm (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasmic staining across tissues (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04451-1) | |
| Caveat | Staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Isoform 2 enriched in testis (UniProt) | |
| Isoform / epitope | 2 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended AKAP1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with three published paraffin-section AKAP1 IHC methods (PMC5520900; PMC6006472; PMC12062205).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A04451-1) |
| Fixation | Image fixative and duration unreported (datasheet A04451-1); 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-AKAP1, 1:50-1:200 (datasheet A04451-1) |
| 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 | AKAP1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 retrieval at 95–98 °C for 20 min (page antigen-retrieval rule); use each article’s stated conditions when reproducing its method.
Section 2
What Is the Expected AKAP1 Staining Pattern?
AKAP1 should appear as granular cytoplasmic staining consistent with mitochondria, particularly in the HPA-listed high-staining cell populations (HPA: granular cytoplasmic expression in all tissues; listed cells High). UniProt places AKAP1 at the mitochondrial outer membrane and reports no transmembrane segment (UniProt Q92667: subcellular location and topology). Interpret the tissue pattern with the HPA’s Supported reliability rating and its stated medium consistency between staining and RNA data (HPA: reliability).
| Granular cytoplasmic chromogen in appendix or duodenal glandular cells. | This matches the expected compartment and examples of high-staining cells (HPA: granular cytoplasmic expression; appendix and duodenum glandular cells High). Assess staining within cells, since a chromogenic section does not resolve the mitochondrial outer membrane itself (UniProt Q92667: mitochondrial outer membrane; standard IHC interpretation). |
| Predominantly nuclear staining, or a continuous cell-surface outline, without granular cytoplasm. | That compartment conflicts with the reported mitochondrial location (UniProt Q92667: mitochondrial outer membrane; HPA: granular cytoplasmic expression). Treat it as suspect, then compare the same run with a known-positive tissue and appropriate detection controls (standard IHC practice). |
| Strongest staining appears in an unexpected cell population while an HPA-listed high-staining population is weak. | Consider cross-reactivity or cell-associated endogenous detection activity, but do not classify every other stained cell as false: HPA reports granular expression across tissues (HPA: tissue profile). Check whether the expected population stains and whether the signal persists in a no-primary control (standard IHC practice). |
| Uniform haze across tissue and empty areas obscures intracellular granules. | The pattern cannot establish AKAP1 localization because the expected readout is cellular, granular cytoplasm (HPA: tissue profile). Review blocking, washing, primary-antibody concentration and chromogen development as general IHC variables; use a no-primary control to assess detection background (standard IHC practice). |
| No granular signal in an HPA-listed high-staining population. | A negative result there needs a technical check before biological interpretation (HPA: listed cells High; reliability Supported). Confirm tissue identity and preservation, compare a positive control from the same run, and review the antibody’s validated IHC-P conditions and detection reagents (standard IHC practice). |
💡Expected AKAP1 appearanceCall a section positive when the relevant cells show discernible granular cytoplasmic staining, with strong signal plausible in HPA-listed High populations; isolated nuclear, cell-surface or uniform background staining is suspect (HPA: tissue profile and listed cell levels; UniProt Q92667: mitochondrial outer membrane).
| Compartment and section resolution | AKAP1 is assigned to the mitochondrial outer membrane (UniProt Q92667: subcellular location), while tissue IHC is described as granular cytoplasmic (HPA: tissue profile). Score the visible granules; an outer-membrane rim is not required in a chromogenic section (standard IHC interpretation). |
| Tissue and cell context | HPA reports granular cytoplasmic expression across tissues, with High staining in specified populations including breast and cervix glandular cells, caudate glial cells, and colon endothelial cells (HPA: tissue IHC). Match the cell population as well as the compartment when choosing a positive control. |
| Isoform distribution | UniProt reports two isoforms: isoform 1 in thymus, prostate, testis, ovary, colon and small intestine; isoform 2 is high in testis and lower in kidney, pancreas, liver, lung and brain (UniProt Q92667: tissue specificity). These reports do not predict an isoform-specific IHC pattern without antibody epitope information. |
| Evidence strength and antibody choice | The HPA tissue profile is Supported, with medium staining–RNA consistency (HPA: tissue IHC reliability). HPA008691 and CAB019286 have Supported IHC status; HPA008620 has no stated IHC status in the supplied list (HPA: antibody validation). Validation of one antibody does not establish another antibody’s pattern. |
| IF/ICC Q&A: What should a positive cell show? | Mitochondrial localization is supported in ICC-IF, with images listed for A-431, U-251MG, U2OS, CACO-2 and PC-3 (HPA: subcellular). Expect a mitochondrial distribution rather than a nuclear signal (HPA: main location); interpret IF/ICC on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining control tissue is blank. | The result conflicts with the selected population’s reported level, but does not identify which step failed (HPA: listed cells High). | Check tissue identity, positive-control performance, antibody dilution and detection reagents; review the chosen antibody’s validated IHC-P conditions (standard IHC practice). |
| Nuclei dominate the signal. | Nuclear dominance does not match granular cytoplasm or mitochondrial localization (HPA: tissue profile; UniProt Q92667: subcellular location). | Compare with an HPA-listed high-staining population and a no-primary control; reassess antibody specificity if the nuclear pattern persists (standard IHC practice). |
| A continuous cell-surface rim dominates. | The observed outline does not match HPA’s granular cytoplasmic tissue pattern (HPA: tissue profile). | Inspect cell-associated granules at suitable magnification and compare serial positive and detection-control sections before scoring the rim as AKAP1 (standard IHC practice). |
| Chromogen appears in many unrelated cells or tissue-free areas. | Diffuse deposition can reflect detection background or excessive development; tissue-free signal cannot identify AKAP1 (standard IHC practice). | Run a no-primary control, check blocking and washing, and adjust detection or development conditions if the control also stains (standard IHC practice). |
| One expected high-staining cell population is weak while others stain. | HPA levels describe observed populations, and its tissue profile has medium staining–RNA consistency; differences need contextual review (HPA: tissue IHC reliability and listed levels). | Verify the sampled cell population and compare control tissue processed in the same run; avoid declaring loss of AKAP1 from intensity alone (standard IHC interpretation). |
| Results differ between antibodies. | The supplied HPA list gives different IHC validation statuses, and no epitope mapping is supplied to explain a discrepancy (HPA: antibody validation). | Compare staining in the same HPA-listed high-staining cells, using each antibody’s validated IHC-P conditions and appropriate controls (HPA: listed levels; standard IHC practice). |
Experiment evidence
IHC & IF images showing AKAP1 staining
A04451-1 · IHCImmunohistochemistry (IHC) analyzes of AKAP 1 (N306) pAb in paraffin-embedded human brain tissue.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000121057-AKAP1/tissue/cerebral+cortex#imid_2635108
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000121057-AKAP1/tissue/colon#imid_2635034
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000121057-AKAP1/tissue/liver#imid_5001339
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000121057-AKAP1/tissue/kidney#imid_2635020
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB019286; green = AKAP1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB019286; green = AKAP1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AKAP1 IHC & IF
🧪Run appendix first: glandular cells should stain strongly (HPA: High in appendix glandular cells). HPA detects AKAP1 in all 45 scored tissues, so there is no supported negative tissue; use no-primary and isotype controls, and treat cells without specific mitochondrial-pattern staining on the positive slide as background comparators rather than confirmed AKAP1-negative cells (HPA: detected in all 45 tissues; HPA subcellular: mitochondria supported).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: AKAP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AKAP1 in A-431, U-251MG, U2OS, CACO-2, PC-3, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Run a secondary-only section and a control immunoglobulin matched to the primary antibody’s host species, class, clonality, and concentration; an AKAP1-knockout specimen, if available, provides a biological specificity control. For appendix IHC, quench endogenous peroxidase and check background from endogenous biotin if using avidin–biotin detection (standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A04451-1 paraffin-section brain caption does not state a fixative (selected tissue-IHC caption). Antigen retrieval requirements are unreported in the supplied evidence and need empirical optimization; neither frozen sections nor IF are established as easier, although IF can help assess mitochondrial localization (HPA subcellular: mitochondria supported). In appendix, glandular architecture and endogenous enzyme background can complicate interpretation of diffuse chromogen staining (standard chromogenic IHC practice).
HPA tissue IHC evidence for AKAP1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: AKAP1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced AKAP1 IHC Tips
Troubleshoot AKAP1 staining in paraffin sections by checking retrieval, compartment-specific signal, cellular context and scoring before interpreting chromogenic IHC.
How should I adjust retrieval when AKAP1 staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (this page’s retrieval setting). If staining remains weak, compare an adjacent section using a longer exposure to the same buffer while keeping antibody and detection conditions constant; assess whether tissue structure or background deteriorates. AKAP1 is assigned to the mitochondrial outer membrane, with no annotated transmembrane segment (UniProt Q92667 topology), so diffuse staining alone does not establish successful retrieval. Use a positive control section in each run, and compare its granular cytoplasmic pattern with the tissue under study (HPA: granular cytoplasmic expression in all tissues).
Could fixation explain weak or uneven AKAP1 staining?
Target-specific sensitivity of AKAP1 to fixation is unknown from the supplied evidence; the catalog image identifies paraffin-embedded human brain but does not state its fixative (A04451-1 caption). Record the fixative, fixation duration and processing history for each section, then compare sections processed alike before changing retrieval or antibody conditions. For routine formalin-fixed material, inconsistent fixation can alter antigen access as a general IHC concern, so check morphology and the staining of a control section from the same run (standard IHC practice). Do not attribute a fixation effect to AKAP1 solely from its outer-membrane location or phosphorylation sites (UniProt Q92667 topology and modified residues).
What AKAP1 staining pattern should I expect in chromogenic IHC?
Expect granular cytoplasmic staining rather than a uniform nuclear signal: the tissue IHC profile reports granular cytoplasmic expression in all tissues (HPA: tissue profile). This is consistent with AKAP1 at the mitochondrial outer membrane and on its cytoplasmic face (UniProt Q92667 subcellular location and function). Examine staining within identifiable cells at high magnification and compare it with a matched positive control; broad extracellular pigment or staining limited to cut edges should prompt a background check. The paraffin-embedded human brain image demonstrates tissue IHC for the catalog antibody, but its caption does not identify a particular positive brain cell type (A04451-1 caption).
Can a negative section exclude AKAP1 when isoforms or epitopes differ?
A negative section cannot exclude all AKAP1 forms without knowing which sequence the antibody recognizes: the record lists 2 isoforms (UniProt Q92667 isoforms). Isoform 1 has been detected in thymus, prostate, testis, ovary, colon and small intestine, whereas isoform 2 is highly expressed in testis and detected at lower levels in several other tissues (UniProt Q92667 tissue specificity). Check the catalog antibody’s stated immunogen or epitope against both sequences before assigning an isoform-specific result. If the epitope is unavailable, report staining as AKAP1 immunoreactivity and interpret a negative result alongside retrieval performance, tissue preservation and a run control.
How can IF help resolve an ambiguous AKAP1 IHC pattern?
Use IF as a separate follow-up to test whether the IHC granules align with mitochondria; the subcellular reference supports mitochondrial localisation (HPA: subcellular location). Multiplex AKAP1 with a mitochondrial marker and, where examining vascular cells, an endothelial marker to identify the expected cell population (HPA: high staining in cerebral-cortex endothelial cells). Choose a far-red fluorophore when tissue autofluorescence compromises shorter wavelengths, and include single-stain controls to assess bleed-through (standard IF practice). Because AKAP1 faces the cytoplasm at the outer mitochondrial membrane, begin with mild permeabilisation and avoid extracting cellular structures unnecessarily (UniProt Q92667 function and topology); do not treat IF concordance alone as proof of antibody specificity.
How do I reduce diffuse brown signal without losing AKAP1 granules?
First distinguish diffuse background from the reported granular cytoplasmic tissue pattern (HPA: tissue profile). On adjacent sections, compare primary-antibody omission and the full stain to reveal signal arising from detection reagents; include a peroxide block before peroxidase-based DAB development as a general chromogenic IHC step (standard IHC practice). Check whether excess chromogen development, inadequate washing or tissue-edge staining explains the apparent spread, changing 1 variable per comparison. If the control granules weaken along with background, restore the previous condition and adjust blocking or detection instead; the supplied catalog caption gives no antibody dilution to substitute here (A04451-1 caption).
How should I score AKAP1 IHC across heterogeneous cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, because the reported high-staining cells differ across tissues, including glandular, glial and endothelial cells (HPA: tissue positives). For each region, record the percentage of positive cells and an intensity-based H-score using 0–3 intensity grades, or measure positive-cell density per mm² when cell counts are more informative (standard IHC scoring practice). Normalise cell-based scores to the number of eligible, intact cells and area-based counts to the evaluated viable tissue area. Apply one threshold and imaging or scoring procedure across sections, and exclude folds, edges and necrotic regions before comparing groups (standard IHC practice).
When is an AKAP1-positive IHC result credible rather than artefactual?
A credible result shows cell-associated granular cytoplasmic staining, consistent with the tissue profile and mitochondrial assignment (HPA: tissue profile; UniProt Q92667 subcellular location). Check that the stained cell type is identifiable: HPA reports high staining in caudate glial cells and cerebral-cortex endothelial cells, among other populations, but neither observation identifies every stained brain cell (HPA: tissue positives). Treat dominant nuclear or extracellular signal, staining confined to section edges or necrosis, and colour persisting without primary antibody as reasons to investigate artefact (standard IHC practice). Recheck peroxidase blocking when unexplained DAB persists, and confirm interpretation against morphology, controls and the same run’s positive section (standard IHC practice).
Boster reagents
Best AKAP1 / A-kinase anchor protein 1, mitochondrial IHC Antibodies
A04451-1 has real IHC data from paraffin-embedded human brain tissue (catalog IHC image caption); IF is listed for Human, Mouse and Rat (catalog: applications, reactivity).
Real IHC data
Anti-AKAP 1 (N306) Antibody
Cat # A04451-1
A04451-1 is listed for IHC and IF in Human, Mouse and Rat (catalog: applications, reactivity). Its IHC image shows paraffin-embedded human brain tissue (catalog IHC image caption); no IF image is supplied (catalog: IF image list).
Which to pick: For tissue IHC, choose A04451-1: its own image documents a paraffin-embedded human brain section, and its listed IHC dilution is 1:50–1:200 (catalog IHC image caption; catalog: IHC dilution). For IF/ICC, A04451-1 lists IF at 1:50–1:200; ICC validation and an IF image are unreported (catalog: applications, IF dilution, IF image list). For cross-species work, A04451-1 is a rabbit polyclonal antibody listed as reactive with Human, Mouse and Rat; the fixative used for the pictured section is unreported (catalog: host, IHC image caption, reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92667 (AKAP1_HUMAN, A-kinase anchor protein 1, mitochondrial).
- Human Protein Atlas. AKAP1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. AKAP1 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. AKAP1 antibody validation summary (3 antibodies).
- Mitochondrial AKAP1 supports mTOR pathway and tumor growth. Cell death & disease 2017 — PMC5520900.
- Higher expression of A-kinase anchoring-protein 1 predicts poor prognosis in human hepatocellular carcinoma. Oncology letters 2018 — PMC6006472.
- Mitochondrial PKA Is Neuroprotective in a Cell Culture Model of Alzheimer's Disease. Molecular neurobiology 2021 — PMC8260456.
- Hepatic AKAP1 deficiency exacerbates diet-induced MASLD by enhancing GPAT1-mediated lysophosphatidic acid synthesis. Nature communications 2025 — PMC12062205.
- PubMed PMID:8769136 — UniProt-cited evidence.
- PubMed PMID:7499250 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AKAP1 (A-kinase anchor protein 1, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04451-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A04451-1); 2. Fixation: Image fixative and duration unreported (datasheet A04451-1); 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-AKAP1, 1:50-1:200 (datasheet A04451-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04451-1); Caveat: Staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; check antibody epitope coverage (UniProt); Regulation: Isoform 2 enriched in testis (UniProt). Evidence: UniProt Q92667, HPA tissue IHC · Supported, HPA ICC-IF · mitochondria., Datasheet A04451-1, PMC5520900, PMC6006472. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AKAP1/immunohistochemistry-ihc-guide.