This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
ADCK1 / AarF domain-containing protein kinase 1 · IHC design guide
Design Immunohistochemistry for ADCK1
Plan chromogenic ADCK1 IHC in paraffin sections with a catalog antibody dilution of 1:100–1:300 (datasheet A14988-1). Interpret the observed cytoplasmic tissue staining (HPA tissue IHC) alongside the mitochondrial annotation (UniProt), while accounting for uncertain staining reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ADCK1 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 staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A14988-1) | |
| Positive control | Appendix+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) | |
| Caveat | Staining awaits external verification (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended ADCK1 IHC & IF Protocols
The catalog antibody’s IHC protocol (datasheet A14988-1) is followed by a published ADCK1 IHC protocol for mouse sections and human colorectal tissue (PMC8024291).
| Sample | Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A14988-1) |
| Fixation | Image fixative and duration unreported (datasheet A14988-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A14988-1); 20 min, 95–100 °C (standard) |
| 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-ADCK1, 1:100-1:300 (datasheet A14988-1) |
| 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 | ADCK1-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 9.0 retrieval for the catalog antibody (datasheet A14988-1); the published protocol used EDTA at 100 °C (PMC8024291).
Section 2
What Is the Expected ADCK1 Staining Pattern?
ADCK1 is annotated in mitochondria and has no transmembrane segment (UniProt Q86TW2 topology). In paraffin IHC, expect cytoplasmic staining in selected epithelial and glandular cells, including colon glandular cells and kidney tubule cells, both reported at medium intensity (HPA tissue IHC). Treat this as a provisional pattern: HPA rates its tissue IHC reliability uncertain, pending external verification (HPA tissue IHC).
| Cytoplasmic chromogenic staining is visible in colon glandular cells or kidney tubule cells (HPA tissue IHC). | This fits the reported medium staining in those cells (HPA tissue IHC). Score the specified cells and compartment separately from surrounding tissue. A chromogenic cytoplasmic pattern cannot by itself resolve individual mitochondria, although mitochondrial localisation is annotated (UniProt Q86TW2). |
| Predominantly nuclear staining replaces the expected cytoplasmic pattern in a tissue section (HPA tissue IHC). | Flag a compartment mismatch and review controls and antibody specificity before calling the IHC result positive. HPA reports nucleoplasm in ICC-IF, but labels that location uncertain; UniProt annotates mitochondrion (HPA subcellular; UniProt Q86TW2). The IF observation does not establish a nuclear IHC pattern. |
| Strong staining appears in adipocytes or bone-marrow hematopoietic cells, where staining was not detected (HPA tissue IHC). | Compare a known-positive section processed in the same run before interpreting this as ADCK1. Unexpected staining could reflect cross-reactivity or endogenous detection activity (general IHC practice); HPA's uncertain IHC reliability limits any tissue-specific conclusion (HPA tissue IHC). |
| Color is diffuse across cells and tissue spaces, with no clear cell boundaries or compartment. | Treat the result as background rather than a scorable ADCK1 pattern (general IHC practice). Review primary-antibody concentration, blocking, washes and detection-only control; a genuine positive should be assignable to the reported cell type and cytoplasm (HPA tissue IHC). |
| No staining appears in colon glandular cells or kidney tubule cells, both reported at medium intensity (HPA tissue IHC). | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
💡Expected ADCK1 appearanceCall a section provisionally positive when discernible cytoplasmic staining occurs in the specified cells at approximately medium intensity, as reported for colon glandular cells or kidney tubules; diffuse color or staining confined to unexpected cells is suspect (HPA tissue IHC; general IHC practice).
| How should localisation guide scoring? | Use the HPA cytoplasmic tissue pattern for paraffin IHC and record nuclear staining separately (HPA tissue IHC). Mitochondrial localisation is a biological annotation, not proof that individual mitochondria must be visible with chromogenic detection (UniProt Q86TW2; general IHC practice). |
| How strong is the IHC evidence? | HPA calls tissue IHC uncertain and pending external verification; both listed antibodies have uncertain IHC status (HPA tissue IHC; HPA antibodies). Use the reported pattern as a comparison point, and interpret agreement alongside section controls rather than as independent validation. |
| Could isoforms change interpretation? | UniProt lists three ADCK1 isoforms and a protein kinase domain at residues 155–467 (UniProt Q86TW2). The payload does not locate either antibody's epitope or establish isoform-specific IHC staining, so a section cannot be assigned to an isoform from this pattern. |
| Do topology or processing predict staining loss? | UniProt reports no transmembrane segment, signal peptide or propeptide, and no annotated glycosylation sites (UniProt Q86TW2). These annotations do not establish an antigen-retrieval requirement or a fixation effect; choose retrieval conditions from the validated IHC method and controls (general IHC practice). |
| What does the IF/ICC observation mean? | HPA reports nucleoplasm as an uncertain ICC-IF location, while UniProt annotates mitochondrion (HPA subcellular; UniProt Q86TW2). Assess IF/ICC localisation in its own guide; it does not supply a protocol option or override the cytoplasmic tissue IHC observation (HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported positive cells show no chromogenic signal (HPA tissue IHC). | The staining run may have failed, or retrieval and antibody conditions may be unsuitable (general IHC practice); ADCK1-specific fixation sensitivity is unreported in the supplied sources. | Check a run control and detection reagents, then compare retrieval and primary-antibody conditions with the IHC-validated method. Record the negative result without inferring absent protein. |
| Nuclei dominate the IHC result. | The compartment conflicts with HPA's cytoplasmic tissue profile; HPA's nuclear ICC-IF location is uncertain (HPA tissue IHC; HPA subcellular). | Check counterstain versus chromogen, inspect detection-only and positive controls, and report the compartment mismatch. Do not score nuclear color as confirmed ADCK1 IHC. |
| Adipocytes or bone-marrow hematopoietic cells stain strongly (HPA tissue IHC). | HPA reports staining as not detected in those cells; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Compare the same run's reported positive cells, inspect a detection-only control, and reassess antibody concentration before assigning specificity. |
| Color is widespread without a cell-specific pattern. | Nonspecific antibody binding, inadequate washes or detection background can produce diffuse color (general IHC practice). | Review blocking, washes, primary-antibody dilution and detection-only control. Score only signal that can be assigned to a cell type and compartment. |
| A weak signal appears in a proposed negative section. | HPA's not-detected calls are observations under its assay conditions, and its tissue IHC reliability is uncertain (HPA tissue IHC). | Describe the observed cells and intensity, compare controls, and avoid treating an HPA not-detected tissue as proof of absolute absence. |
| Two antibodies give different tissue patterns. | Neither listed antibody has enhanced IHC validation; both have uncertain IHC status (HPA antibodies). | Document each antibody's cell types and compartments against the same controls. Treat disagreement as unresolved specificity rather than selecting a pattern solely by signal strength. |
Experiment evidence
IHC & IF images showing ADCK1 staining
A14988-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000063761-ADCK1/tissue/cerebral+cortex#imid_14486916
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000063761-ADCK1/tissue/colon#imid_14487554
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000063761-ADCK1/tissue/liver#imid_14487555
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000063761-ADCK1/tissue/kidney#imid_14487564
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA055014; green = ADCK1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA055014; green = ADCK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ADCK1 IHC & IF
🧪Run appendix first and assess its glandular cells for staining (HPA: Medium in appendix glandular cells). Use adipose tissue adipocytes as a negative comparator (HPA: Not detected in adipocytes); adjacent nonglandular cells on the appendix slide should show only counterstain or background before they are treated as internal negatives, since their ADCK1 status is not specified by the supplied HPA row (HPA: appendix glandular-cell annotation).
Positive control tissue: Appendix (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADCK1 in A-431, MCF-7, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (uncertain) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control and an isotype control matched to the primary antibody’s host species and immunoglobulin class, with the control format appropriate to its clonality (standard IHC practice). Confirm specificity with ADCK1-knockout material or a peptide-blocking control if the immunizing peptide is available, and quench endogenous peroxidase for chromogenic detection, especially when evaluating inflammatory cells in appendix (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (A14988-1 caption: fixative unreported). The caption uses Tris-EDTA at pH 9.0 for antigen retrieval and a 1:200 primary dilution overnight at 4°C; it does not establish that retrieval is required (A14988-1 caption). Whether frozen sections or IF are easier is unreported; assess appendix inflammatory-cell peroxidase background in IHC (standard IHC practice), and interpret IF localisation cautiously because HPA calls nucleoplasmic localisation uncertain while UniProt lists mitochondrion (HPA: Nucleoplasm, uncertain; UniProt Q86TW2: Mitochondrion).
HPA tissue IHC evidence for ADCK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Pending external verification.). 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 | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | Low | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADCK1 IHC Tips
Troubleshoot ADCK1 staining by optimizing paraffin-section conditions and checking cell type and subcellular pattern against independent evidence.
Which retrieval condition should I try first for weak ADCK1 staining in paraffin sections?
Start with heat-mediated Tris-EDTA at pH 9.0 for ADCK1 antigen retrieval (datasheet A14988-1). The paraffin-section example used this buffer, then primary antibody at 1:200 overnight at 4°C (caption A14988-1). If staining remains weak, compare a shorter and longer heating cycle on matched sections while holding detection conditions constant (standard IHC practice). If that fails, test an alternative retrieval buffer as a fallback, recording both signal and tissue damage (standard IHC practice). Judge improvement in identifiable cells rather than by stronger diffuse color alone; the published tissue staining profile has uncertain reliability (HPA: tissue IHC reliability Uncertain).
How should I troubleshoot fixation when ADCK1 staining varies between paraffin blocks?
Target-specific fixation sensitivity is unknown from the supplied ADCK1 evidence; the selected paraffin-section caption does not state a fixative (caption A14988-1). Record each block’s fixative, fixation duration, processing history, and section age before attributing a difference to ADCK1 expression (standard IHC practice). Compare blocks using the same pH 9.0 retrieval and 1:200 primary dilution documented for the catalog example (caption A14988-1). Include a consistently stained reference section in each run and inspect morphology for overprocessing or tissue loss (standard IHC practice). Do not infer a fixation effect from ADCK1 topology, modification records, or tissue staining patterns (UniProt Q86TW2; HPA: tissue IHC reliability Uncertain).
Is diffuse cytoplasmic or nuclear chromogen compatible with ADCK1 localisation?
ADCK1 is annotated as mitochondrial, without a transmembrane segment (UniProt Q86TW2: subcellular location and topology). At chromogenic IHC resolution, evaluate whether cytoplasmic staining is granular and cell-associated; avoid calling individual mitochondria from a diffuse brown field (standard IHC practice). HPA describes cytoplasmic tissue staining but assigns its tissue IHC profile uncertain reliability (HPA: cytoplasmic expression in several tissues; reliability Uncertain). HPA’s nucleoplasmic call comes from subcellular IF and is also uncertain, so nuclear-only chromogen needs independent confirmation (HPA: nucleoplasm, uncertain). Compare candidate staining with hematoxylin-defined cell boundaries and a no-primary control before assigning a compartment (standard IHC practice).
Could isoform choice or epitope accessibility explain inconsistent ADCK1 staining?
ADCK1 has 3 annotated isoforms and a protein kinase domain at residues 155–467 (UniProt Q86TW2: isoforms and domains). The supplied caption does not identify the catalog antibody’s epitope, so staining cannot be assigned to one isoform from this IHC result (caption A14988-1). Check the antibody’s immunogen coordinates against isoform sequences before comparing blocks or claiming isoform-specific expression (standard IHC practice). If an independent antibody recognizes a distinct shared region, compare cell type and compartment patterns on adjacent sections under separately optimized conditions (standard IHC practice). Treat discordance as a reason to test epitope accessibility and antibody specificity, rather than as proof of altered isoform abundance (standard IHC practice).
How can IF help resolve ambiguous ADCK1 chromogenic staining?
Use IF as a separate confirmation experiment: multiplex ADCK1 with a marker for the expected cell type, such as kidney tubular cells when examining kidney (HPA: medium staining in kidney tubule cells). Choose spectrally separated fluorophores and inspect unstained tissue for autofluorescence before interpreting overlap (standard IF practice). ADCK1 is mitochondrial and has no annotated transmembrane segment, but the catalog epitope’s membrane-facing side is unspecified (UniProt Q86TW2: location and topology; caption A14988-1: epitope unreported). Optimize permeabilisation for access to that epitope, with matched controls for morphology and background (standard IF practice). Do not carry the paraffin-section retrieval or dilution into IF without validation (caption A14988-1: paraffin-section conditions).
What should I change if ADCK1 DAB staining is widespread or patchy?
First compare a no-primary control with the ADCK1 section to locate detection-system or endogenous-enzyme background (standard IHC practice). For peroxidase-based DAB detection, verify the peroxidase block, wash consistency, and chromogen development time; these are general workflow controls (standard IHC practice). The catalog example used primary antibody at 1:200 overnight at 4°C and secondary at 1:200 for 30 minutes at room temperature (caption A14988-1). If controls are clean but tissue remains uniformly brown, titrate primary concentration and shorten development on matched sections (standard IHC practice). Exclude folds, dried edges, and damaged areas before judging cytoplasmic signal (standard IHC practice).
How should I score ADCK1 staining across samples with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and intensity with an H-score where morphology permits (standard IHC practice). Normalize the denominator to evaluable cells of the specified type, not total tissue area when sample composition differs (standard IHC practice). For spatial questions, report positive-cell density per mm² of viable, annotated tissue and state whether stromal cells were included (standard IHC practice). Keep retrieval, detection, exposure to DAB, and score thresholds consistent across slides (standard IHC practice). Because HPA rates its ADCK1 tissue IHC profile uncertain, validate scoring thresholds with controls rather than treating its intensity categories as calibrated standards (HPA: reliability Uncertain).
How do I distinguish convincing ADCK1 staining from a false-positive pattern?
Prioritize reproducible, cell-associated cytoplasmic staining in morphologically intact cells, consistent with mitochondrial ADCK1 and reported cytoplasmic tissue staining (UniProt Q86TW2: mitochondrion; HPA: cytoplasmic tissue expression). A nuclear-only pattern warrants scrutiny because the HPA nucleoplasmic IF assignment is uncertain (HPA: nucleoplasm, uncertain). Check whether the stained cell type matches the claim: HPA reports medium signal in kidney tubule cells but no detection in bone-marrow hematopoietic cells (HPA: tissue IHC profile, reliability Uncertain). Discount staining confined to cut edges, necrosis, or folds, and compare with a no-primary control for endogenous peroxidase or detection artefact (standard IHC practice). Confirm consequential findings with independent specificity evidence (standard IHC practice).
Boster reagents
Best ADCK1 / AarF domain-containing protein kinase 1 IHC Antibodies
A14988-1 has IHC data from paraffin-embedded human tonsil (IHC image caption). IF is listed for the human- and mouse-reactive antibody, but no IF image is supplied (catalog: applications, reactivity, IF image alts).
Real IHC data
Anti-ADCK1 Antibody
Cat # A14988-1
A14988-1 will render with its paraffin-section human tonsil IHC figure (IHC image caption). The antibody lists IHC and IF applications and human and mouse reactivity; IF has no supplied figure (catalog: applications, reactivity, IF image alts).
Which to pick: For tissue IHC, choose A14988-1: its caption documents paraffin-embedded human tonsil, antibody at 1:200 overnight at 4 °C, and Tris-EDTA retrieval at pH 9.0 (A14988-1 IHC image caption). For IF, A14988-1 lists a 1:200–1:1000 dilution; ICC validation and an IF figure are unreported (catalog: applications, IF dilution, IF image alts). For cross-species work, A14988-1 lists human and mouse reactivity, though its IHC figure shows only human tonsil; clonality and the tissue fixative are unreported (catalog: reactivity, clone field; A14988-1 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q86TW2 (ADCK1_HUMAN, AarF domain-containing protein kinase 1).
- Human Protein Atlas. ADCK1 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ADCK1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. ADCK1 antibody validation summary (2 antibodies).
- ADCK1 activates the β-catenin/TCF signaling pathway to promote the growth and migration of colon cancer cells. Cell death & disease 2021 — PMC8024291.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12508121 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ADCK1 (AarF domain-containing protein kinase 1). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A14988-1). Validated IHC antibody: A14988-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil tissue; fixative not specified (datasheet A14988-1); 2. Fixation: Image fixative and duration unreported (datasheet A14988-1); 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 retrieval: Tris-EDTA pH 9.0 (datasheet A14988-1); 20 min, 95–100 °C (standard); 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-ADCK1, 1:100-1:300 (datasheet A14988-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining awaits external verification (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope differences unspecified (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q86TW2, HPA tissue IHC · Uncertain, HPA ICC-IF · nucleoplasm., Datasheet A14988-1, PMC8024291. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ADCK1/immunohistochemistry-ihc-guide.