This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MINK1 / Misshapen-like kinase 1 · IHC design guide
Design Immunohistochemistry for MINK1
High staining in lung macrophages and colon glandular cells (HPA tissue IHC) helps select positive controls. Start the catalog antibody at 1:100–1:300 for paraffin IHC (datasheet A30524).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MINK1 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 signal in colon glands and lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30524) | |
| 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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A30524) | |
| Caveat | Strong lung macrophage staining may affect scoring (HPA tissue IHC) | |
| Regulation | Isoform abundance varies by tissue (UniProt) | |
| Isoform / epitope | 5 isoforms; no propeptide; check epitope coverage (UniProt) |
Section 1
Recommended MINK1 IHC & IF Protocols
The catalog antibody protocol is paired with one published MINK1 IHC protocol using lung sections (PMC9592806).
| Sample | Paraffin-embedded Human breast cancer tissue; fixative not specified (datasheet A30524) |
| Fixation | Image fixative and duration unreported (datasheet A30524); 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 8.0 (datasheet A30524); 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-MINK1, 1:100 - 1:300 (datasheet A30524) |
| 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 | MINK1-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA retrieval at pH 8.0 (datasheet A30524); the published lung protocol does not specify retrieval (PMC9592806).
Section 2
What Is the Expected MINK1 Staining Pattern?
MINK1 is primarily cytoplasmic in tissue IHC, with high staining reported in selected glandular, squamous epithelial, macrophage and trophoblastic cells (HPA: cytoplasmic expression in several tissues; HPA: High in listed cells). UniProt also places MINK1 at the Golgi and neuronal projections, and reports no transmembrane segment (UniProt Q8N4C8 subcellular location; topology). The HPA tissue IHC assessment is Approved, with medium consistency between staining and RNA expression (HPA: reliability).
| Cytoplasmic staining is strong in colon or appendix glandular cells, cervical squamous epithelial cells, lung macrophages, or placental trophoblasts (HPA: High in these cells). | This matches observed positive cell types and the reported cytoplasmic pattern (HPA: tissue IHC profile). Compare the stained cell population with tissue morphology; a positive call should describe the cells and compartment, not just the tissue name. High is the HPA observation for those specific cells, not a required intensity in every specimen (HPA: tissue IHC). |
| Signal is confined to nuclei or traces crisp cell borders while the cytoplasm is clear. | A predominantly nuclear or membrane-outline pattern conflicts with the reported tissue pattern and a protein without a transmembrane segment (HPA: cytoplasmic expression; UniProt Q8N4C8 topology). Check morphology, counterstain and detection controls before calling it MINK1. Golgi or neuronal-projection localisation is annotated, but neither annotation establishes a distinctive chromogenic pattern here (UniProt Q8N4C8 subcellular location). |
| Strong staining appears mainly in adipocytes, cardiomyocytes, or bone-marrow hematopoietic cells. | These particular cell populations were not detected by HPA tissue IHC (HPA: Not detected in adipocytes, cardiomyocytes and bone-marrow hematopoietic cells). Unexpected signal raises possible nonspecific antibody binding or detection activity; inspect a no-primary control and the positive tissue on the same run. An HPA negative observation is a comparison point, not proof that every sample must be negative (HPA: Approved; medium consistency). |
| Brown color spreads across stroma, empty spaces, or most cell types without clear cytoplasmic boundaries. | Treat this as background until morphology separates cellular staining from deposits. In chromogenic IHC, incomplete blocking, excess reagent or residual endogenous detection activity can produce broad color (standard IHC practice). The HPA description is cytoplasmic expression in several tissues, so diffuse color alone does not identify MINK1-positive cells (HPA: tissue IHC profile). |
| No staining is visible in a positive control such as colon glandular cells or lung macrophages. | HPA reports high staining in those defined cell populations (HPA: High in colon glandular cells; High in lung macrophages). If the expected cells are present, first suspect a run-level problem with antibody, retrieval or detection (standard IHC practice). Do not infer biological absence from a failed control; the supplied evidence does not establish MINK1-specific fixation sensitivity. |
💡Expected MINK1 appearanceCall a section positive when identifiable glandular, squamous epithelial, macrophage or trophoblastic cells show clear cytoplasmic staining, potentially strong in HPA-listed high-staining populations; isolated nuclear, border-only or acellular color needs control review (HPA: cytoplasmic profile and High cell populations; UniProt Q8N4C8 topology).
| Which tissue and cell population are scored | HPA reports High staining in appendix, colon and duodenum glandular cells; cervix, esophagus and oral-mucosa squamous epithelial cells; lung macrophages; and placental trophoblasts (HPA: tissue IHC). It reports Not detected for selected other cell populations, including adipocytes and cardiomyocytes (HPA: tissue IHC). Score the named cells within each section; whole-tissue labels can obscure a mixed result. |
| Expected compartment and resolution | Cytoplasm is the documented tissue IHC pattern (HPA: tissue IHC profile). Golgi, axonal, dendritic and postsynaptic-density locations are UniProt annotations (UniProt Q8N4C8 subcellular location). Routine chromogenic sections may not resolve those structures, so do not require visible puncta to accept cytoplasmic staining (standard IHC practice). |
| Isoforms and antibody epitope | MINK1 has five listed isoforms; isoform 3 is ubiquitous, while isoform 1 is most abundant in skeletal muscle and isoform 2 exceeds isoform 1 in brain (UniProt Q8N4C8 tissue specificity; isoforms). The supplied antibody record gives no epitope or isoform coverage (HPA: HPA056296 validation record). Staining cannot therefore be assigned to a particular isoform from this evidence. |
| Strength of antibody and tissue evidence | The listed antibody HPA056296 has Approved IHC status; the tissue profile has medium staining–RNA consistency (HPA: antibody validation; tissue reliability). Neither entry reports Enhanced IHC validation (HPA: antibody validation). Use the observed pattern as a reference while retaining morphology and run controls when evaluating an unexpected result (standard IHC practice). |
| IF/ICC: what pattern can be expected? | A cytoplasmic location is supported by UniProt, which also annotates Golgi and neuronal projections (UniProt Q8N4C8 subcellular location). HPA supplies no main ICC-IF location and no cell-line ICC-IF images, so this payload cannot establish a verified fluorescent distribution (HPA: subcellular summary). Interpret any IF/ICC result on its own guide page. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive control is blank. | An antibody, retrieval or detection step may have failed (standard IHC practice). | Verify that the expected glandular cells or macrophages are present, then check the run reagents and detection controls (HPA: High in colon glandular cells and lung macrophages; standard IHC practice). |
| Only nuclei or cell borders stain. | The dominant compartment disagrees with the reported cytoplasmic tissue pattern (HPA: tissue IHC profile). | Review morphology and a no-primary control; repeat the stain if the pattern persists, and report the compartment actually seen (standard IHC practice). |
| Unexpected cells stain strongly in a comparison tissue. | Cross-reactivity or detection activity is possible; HPA's negative calls apply to named cells, not every cell in the organ (HPA: tissue IHC). | Identify the stained cell type and compare it with an HPA-listed positive population and a no-primary control before assigning MINK1 (HPA: tissue IHC; standard IHC practice). |
| The whole section has diffuse brown background. | Excess reagent, incomplete blocking or endogenous detection activity can obscure cellular signal (standard IHC practice). | Check no-primary and detection controls, then adjust blocking, washing or reagent concentration according to the validated IHC workflow (standard IHC practice). |
| Lung macrophages are dark in both test and no-primary sections. | A detection-system contribution is plausible when color remains without primary antibody (standard IHC practice). | Resolve the control signal before using macrophages as evidence of MINK1; HPA reports high macrophage staining only as a tissue IHC observation (HPA: High in lung macrophages; standard IHC practice). |
| Brain or skeletal-muscle staining seems inconsistent with expression notes. | UniProt describes tissue-dependent isoform abundance, while HPA reports Low staining in hippocampal neurons and Not detected in specified glial populations (UniProt Q8N4C8 tissue specificity; HPA: tissue IHC). | Score the actual cell population and compartment. Do not convert an isoform-expression note into a predicted IHC intensity without epitope or isoform-coverage evidence (HPA: tissue IHC; HPA: HPA056296 validation record). |
Experiment evidence
IHC & IF images showing MINK1 staining
A30524 · IHCImmunohistochemical analysis of paraffin-embedded Human breast cancer. Antibody was diluted at 1:100(4° overnight). High-pressure and temper
A06134-2 · IHCImmunohistochemistry validation of MINK1 using Anti-Misshapen-like kinase 1 MINK1 Antibody (A06134-2). Immunohistochemical analysis of paraf
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141503-MINK1/tissue/cerebral+cortex#imid_15640811
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141503-MINK1/tissue/colon#imid_15640241
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141503-MINK1/tissue/liver#imid_15640226
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000141503-MINK1/tissue/kidney#imid_15640227
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MINK1 IHC & IF
🧪Run colon first: its glandular cells should stain (HPA: High in colon glandular cells). Use adipose tissue as the biological negative (HPA: Not detected in adipocytes); on the colon slide, cells selected as internal negatives should show only background chromogen, with clear separation from glandular staining.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for MINK1; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.
Technical controls: Include no-primary (secondary-only) and host-species- and antibody-class-matched isotype controls, plus a MINK1 knockout or immunogen-peptide-block control; peptide blocking was used for the selected antibody’s tissue-IHC image (caption: peptide-preabsorbed control). Quench endogenous peroxidase before chromogenic detection and assess residual background in the colon section (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 (caption: fixative unreported). That caption reports high-pressure, high-temperature Tris-EDTA retrieval at pH 8.0 for breast cancer tissue, providing a retrieval starting point rather than evidence that colon staining requires those conditions (caption: Tris-EDTA pH 8.0 retrieval). Whether frozen sections or IF are easier is unreported; assess colon glandular staining against local background because the HPA positive call identifies glandular cells, not every cell in the section (HPA: High in colon glandular cells).
HPA tissue IHC evidence for MINK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | 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 → |
Section 3
Advanced MINK1 IHC Tips
Troubleshoot MINK1 staining in paraffin sections by checking retrieval, cellular pattern, controls and scoring before interpreting chromogenic signal.
Which retrieval conditions should I start with for MINK1 paraffin sections?
Start with heat-mediated retrieval in Tris-EDTA at pH 8.0 (datasheet A30524). The selected paraffin-section image used high pressure and temperature with that buffer, followed by antibody at 1:100 overnight at 4°C (caption A30524). If staining is weak, compare a shorter and longer heating interval on adjacent sections while keeping detection and exposure to chromogen constant (standard IHC practice). Include a positive tissue and an antibody-omission control in each comparison so stronger staining can be distinguished from increased background (standard IHC practice). Do not treat a changed retrieval condition as validated solely because it produces darker signal (standard IHC practice).
Could fixation explain weak or patchy MINK1 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (caption A30524). Record the fixative and fixation duration for each specimen, then compare sections with similar processing before changing the antibody concentration (standard IHC practice). Uneven preservation, drying or detached sections can create patchy chromogenic staining independent of antigen abundance (standard IHC practice). Use the stated Tris-EDTA pH 8.0 retrieval and 1:100 overnight incubation as a documented starting point for this antibody (caption A30524). If the pattern remains inconsistent, test retrieval on matched sections and report the processing difference rather than assigning it to MINK1 biology (standard IHC practice).
Where should convincing MINK1 signal appear in a tissue section?
Assess staining primarily within the cytoplasmic compartment: tissue IHC reports cytoplasmic expression in several tissues (HPA: tissue IHC profile). MINK1 is also annotated in the Golgi apparatus, postsynaptic density, axons and dendrites (UniProt Q8N4C8: subcellular locations). It has no transmembrane segment, so a sharp membrane-only outline needs additional validation before being called MINK1 (UniProt Q8N4C8: topology). Compare the cellular pattern with a positive reference: glandular cells in the appendix and lung macrophages have reported High staining (HPA: appendix; HPA: lung). Keep compartment and cell identity separate in the score, because signal in the expected compartment alone does not establish antibody specificity (standard IHC practice).
Can this stain distinguish MINK1 isoforms or modified epitopes?
Do not assign a positive chromogenic stain to one isoform without an isoform-resolved epitope map or separate validation (standard IHC practice). MINK1 has 5 listed isoforms, with isoform 2 more abundant than isoform 1 in brain and isoform 3 described as ubiquitous (UniProt Q8N4C8: isoforms and tissue specificity). Its kinase domain spans residues 25–289, and its CNH domain spans 1019–1306 (UniProt Q8N4C8: domains). Phosphorylation and methylation sites are annotated, but the supplied antibody evidence does not locate its binding epitope relative to them (UniProt Q8N4C8: modified residues; caption A30524). Treat peptide pre-absorption as support for reagent-dependent staining, not proof of isoform specificity (caption A30524; standard IHC practice).
How should I investigate MINK1 localisation by multiplex immunofluorescence?
Treat IF as a separate application requiring its own antibody and staining validation; the supplied tissue example documents chromogenic staining of paraffin sections (caption A30524). Pair MINK1 with a validated marker for the cell population under study, such as a macrophage marker when examining the reported lung macrophage signal (HPA: lung; standard IF practice). Choose spectrally separated fluorophores and compare unstained tissue in each channel; a far-red channel can help when shorter-wavelength tissue autofluorescence is strong (standard IF practice). MINK1 lacks a transmembrane segment and has intracellular locations, so use a permeabilisation condition that gives antibody access to the intracellular epitope, then check whether morphology is preserved (UniProt Q8N4C8: topology and subcellular locations; standard IF practice). No MINK1 ICC/IF image evidence is supplied here (HPA: subcellular record).
How can I reduce diffuse brown background without losing MINK1 signal?
First inspect an antibody-omission control and a section stained with the same detection reagents to identify nonspecific chromogen deposition (standard IHC practice). In peroxidase-based chromogenic IHC, block endogenous peroxidase before adding the detection reagent and keep DAB development consistent across sections (standard IHC practice). Titrate from the documented 1:100 overnight incubation if background obscures cell borders, changing one variable at a time (caption A30524; standard IHC practice). Compare the result with the peptide-preabsorbed negative control shown for the selected antibody, while recognising that it does not identify every source of background (caption A30524; standard IHC practice). Record whether haze follows tissue folds, edges or damaged areas before interpreting it as cytoplasmic MINK1 (standard IHC practice; HPA: tissue IHC profile).
What is a defensible way to quantify MINK1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive cells and cytoplasmic intensity separately (standard IHC practice; HPA: tissue IHC profile). An H-score can combine intensity categories 0–3 with their percentages to yield 0–300; apply the same thresholds and chromogen development across specimens (standard IHC practice). For a focal population such as lung macrophages, positive-cell density per mm² may be useful if total macrophage density is also measured (HPA: High in lung macrophages; standard IHC practice). Normalise positive cells to the number of evaluable cells of the chosen type, or density to viable tissue area, and exclude folds, necrosis and section edges consistently (standard IHC practice).
Which patterns argue that apparent MINK1 positivity is an artefact?
A reproducible cytoplasmic signal in the expected cell population is more persuasive than brown staining confined to a section edge or necrotic area (HPA: tissue IHC profile; standard IHC practice). Check whether signal localises to reported positive populations, such as lung macrophages or appendix glandular cells, while interpreting reported negatives at the stated cell-type level (HPA: lung; HPA: appendix; HPA: negative-cell entries). A nuclear-only or membrane-only pattern needs separate validation against the reported intracellular locations and absence of a transmembrane segment (UniProt Q8N4C8: subcellular locations and topology). Persistent colour in an antibody-omission control suggests detection chemistry or endogenous enzyme activity rather than specific antibody binding (standard IHC practice). The tissue IHC assessment is Approved with medium staining–RNA consistency, so avoid equating stain intensity with kinase activity (HPA: reliability; standard IHC interpretation).
Boster reagents
Best MINK1 / Misshapen-like kinase 1 IHC Antibodies
Both anti-MINK1 antibodies have IHC images from paraffin-embedded human breast cancer (each SKU’s image caption); both list IF and human/mouse reactivity (catalog entries).
Real IHC data
Anti-MAP4K6 MINK1 Antibody
Cat # A30524
Real IHC data
Anti-Misshapen-like kinase 1 MINK1 Antibody
Cat # A06134-2
A30524 lists IHC, IF and ICC, with an IHC image from paraffin-embedded human breast cancer (A30524 catalog entry and image caption). A06134-2 lists IHC and IF, with its own IHC image from paraffin-embedded human breast cancer (A06134-2 catalog entry and image caption).
Which to pick: Either SKU is supported for paraffin-section IHC by its own human breast cancer image; the fixative is unreported in both captions (A30524 and A06134-2 image captions). Choose A30524 when ICC is required because it lists ICC and is polyclonal; both SKUs list IF, while A06134-2 has no reported clonality (catalog entries). Both list human and mouse reactivity, but their IHC images show human tissue only (catalog entries and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8N4C8 (MINK1_HUMAN, Misshapen-like kinase 1).
- Human Protein Atlas. MINK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MINK1 subcellular location (ICC-IF): Highest expression in HaCaT: 86.9 nTPM.
- Human Protein Atlas. MINK1 antibody validation summary (1 antibodies).
- MBD2 mediates Th17 cell differentiation by regulating MINK1 in Th17-dominant asthma. Frontiers in genetics 2022 — PMC9592806.
- Resveratrol contributes to NK cell-mediated breast cancer cytotoxicity by upregulating ULBP2 through miR-17-5p downmodulation and activation of MINK1/JNK/c-Jun signaling. Frontiers in immunology 2025 — PMC11830804.
- PubMed PMID:10708748 — UniProt-cited evidence.
- PubMed PMID:15469942 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MINK1 (Misshapen-like kinase 1). Expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A30524). Validated IHC antibody: A30524 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 breast cancer tissue; fixative not specified (datasheet A30524); 2. Fixation: Image fixative and duration unreported (datasheet A30524); 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 8.0 (datasheet A30524); 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-MINK1, 1:100 - 1:300 (datasheet A30524); 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 A30524); Caveat: Strong lung macrophage staining may affect scoring (HPA tissue IHC); Isoform / epitope: 5 isoforms; no propeptide; check epitope coverage (UniProt); Regulation: Isoform abundance varies by tissue (UniProt). Evidence: UniProt Q8N4C8, HPA tissue IHC · Approved, HPA cell-line RNA · Highest expression in HaCaT: 86.9 nTPM, Datasheet A30524, PMC9592806. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MINK1/immunohistochemistry-ihc-guide.