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MAP3K6 / Mitogen-activated protein kinase kinase kinase 6 · IHC design guide
Design Immunohistochemistry for MAP3K6
Plan MAP3K6 chromogenic IHC around the ubiquitous cytoplasmic pattern reported in tissue sections (HPA tissue IHC). The catalog antibody lists an IHC dilution of 1:100–1:300 (datasheet), and high staining is reported in respiratory epithelial and endometrial glandular cells (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAP3K6 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 | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Bronchus+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 | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining–RNA agreement is medium; verify the pattern (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope effects are unestablished (UniProt) |
Section 1
Recommended MAP3K6 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by published chromogenic IHC methods for gastric cancer cells (PMC10913707) and rat lung sections (PMC13616702).
| Sample | Paraffin-embedded human breast carcinoma tissue; fixative not specified (datasheet A09063) |
| Fixation | Image fixative and duration unreported (datasheet A09063); 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-MAP3K6, 1:100-1:300 (datasheet A09063) |
| 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 | MAP3K6-positive staining in respiratory epithelial cells of bronchus (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. 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); the rat lung protocol also uses citrate buffer (PMC13616702).
Section 2
What Is the Expected MAP3K6 Staining Pattern?
In paraffin IHC, expect mainly cytoplasmic MAP3K6 staining across tissues, with high staining reported in respiratory epithelium, several glandular cell populations, smooth muscle cells and other listed cells (HPA: tissue IHC). The tissue profile is Approved, with medium consistency between staining and RNA data and external verification pending (HPA: reliability). MAP3K6 has no annotated transmembrane segment or signal peptide (UniProt O95382: topology and processing).
| Cytoplasmic staining in respiratory epithelial cells, pancreatic exocrine cells or smooth muscle cells. | This matches the reported high tissue staining and ubiquitous cytoplasmic profile (HPA: tissue IHC). Compare cell identity and compartment before scoring intensity. |
| Only extracellular or luminal deposits appear, with no cytoplasmic staining in the expected cells. | This is a compartment mismatch and suggests an IHC artefact; the tissue profile is cytoplasmic (HPA: tissue IHC). Check deposits and detection controls before calling it MAP3K6. |
| Strong staining appears only in a cell population outside the reported tissue pattern. | Consider antibody cross-reactivity or endogenous detection activity (general IHC practice). HPA reports low tissue specificity, so an unlisted cell type alone does not establish a false positive (HPA: tissue IHC). |
| Brown haze covers cells and surrounding tissue without clear cell boundaries. | Diffuse background prevents a reliable compartment call (general IHC practice). It cannot be scored as the cytoplasmic pattern reported for tissue IHC (HPA: tissue IHC). |
| No staining appears in a section containing a reported high-staining cell population. | An absent signal in that candidate positive control calls for a run check; it does not establish that the specimen lacks MAP3K6 (HPA: tissue IHC; general IHC practice). |
💡Expected MAP3K6 appearanceCall a positive IHC result when identifiable cells show predominantly cytoplasmic staining, especially the reported high-staining populations; isolated extracellular deposits or uniform haze are suspect (HPA: tissue IHC; general IHC practice).
| Tissue and cell choice | HPA reports high staining in bronchial respiratory epithelium, endometrial and stomach glandular cells, pancreatic exocrine cells, smooth muscle cells, peripheral nerve and cells in seminiferous ducts; adipocytes are medium (HPA: tissue IHC). |
| Strength of evidence | The tissue profile is Approved but has medium staining–RNA consistency and awaits external verification; HPA051192 is IHC Approved, with no Enhanced IHC status supplied (HPA: tissue reliability and antibody status). |
| Compartment and topology | Tissue IHC is described as ubiquitously cytoplasmic (HPA: tissue IHC). UniProt gives no subcellular annotation and reports no transmembrane segment or signal peptide; topology alone cannot settle every stain location (UniProt O95382). |
| Isoforms and epitope scope | Three isoforms are listed, but the supplied record does not map the IHC antibody epitope or establish which isoforms it detects (UniProt O95382: isoforms; HPA: antibody status). |
| IF/ICC Q&A: should it match IHC? | HPA ICC-IF places MAP3K6 mainly in the nucleoplasm, with additional plasma membrane, cytosol and actin filament localization (HPA: subcellular). Interpret that assay separately from cytoplasmic tissue IHC (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a reported high-staining control cell population. | The run may have failed, or the positive cells may be absent from the examined area (general IHC practice). | Confirm cell identity and section integrity; inspect the run controls, retrieval record, primary-antibody step and detection reagents before interpreting a negative (general IHC practice; HPA: tissue IHC). |
| Extracellular or luminal staining dominates while expected cell cytoplasm is clear. | A deposit or other staining artefact may mimic positivity (general IHC practice); this distribution conflicts with the tissue profile (HPA: tissue IHC). | Compare with a no-primary control and a reported high-staining cell population; score only localized cellular staining (general IHC practice; HPA: tissue IHC). |
| Unexpected cells stain more strongly than the reported cell population. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA does not provide a formal negative-cell list (HPA: tissue IHC). | Check morphology, no-primary and detection controls, and a reported positive cell type before assigning specificity (general IHC practice; HPA: tissue IHC). |
| The entire section has diffuse brown background. | Nonspecific reagent binding or incomplete endogenous-enzyme blocking can obscure cellular staining (general IHC practice). | Review blocking, washes and detection controls; reassess whether cytoplasmic signal remains distinguishable (general IHC practice; HPA: tissue IHC). |
| Only nuclei stain in paraffin IHC. | This differs from the cytoplasmic tissue profile, although nucleoplasm is the main approved ICC-IF location (HPA: tissue IHC and subcellular). | Check tissue morphology and IHC controls before calling an artefact or a new IHC pattern; do not transfer the ICC-IF location directly to IHC scoring (HPA: tissue IHC and subcellular; general IHC practice). |
| Weak staining in Purkinje cells, caudate neurons or non-germinal-center lymph-node cells. | These are reported low-staining populations, so weak signal alone is inconclusive (HPA: tissue IHC). | Use a reported high-staining population to judge run performance, then score the low-staining cells by compartment and background (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing MAP3K6 staining
A09063 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, using MAP3K6 Antibody. The picture on the right is blocked
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000142733-MAP3K6/tissue/cerebral+cortex#imid_14602448
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000142733-MAP3K6/tissue/colon#imid_14602382
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000142733-MAP3K6/tissue/liver#imid_14602383
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000142733-MAP3K6/tissue/kidney#imid_14602392
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA051192; green = MAP3K6, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA051192; green = MAP3K6, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAP3K6 IHC & IF
🧪Run bronchus first and look for staining in respiratory epithelial cells (HPA: High in bronchus respiratory epithelial cells). HPA detects MAP3K6 in all 45 scored tissues, so use no-primary and isotype controls for negative signal; any unstained cells on the bronchus slide are background comparators, not validated negative cells (HPA: no negative tissue rows).
Positive control tissue: Bronchus (Respiratory epithelial cells, HPA High)
Negative control tissue: None in HPA: MAP3K6 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 MAP3K6 in HeLa, MCF-7, U2OS, NIH 3T3, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide and an isotype control matched to the primary antibody’s host species and immunoglobulin class; use MAP3K6 knockout material, if available, as a biological negative, and a peptide-blocked section as a specificity check (A09063 tissue-IHC caption: peptide block). For chromogenic bronchus IHC, quench endogenous peroxidase and block endogenous biotin if using avidin–biotin detection; check tissue autofluorescence before interpreting IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval empirically for paraffin sections (A09063 tissue-IHC caption: paraffin-embedded tissue). The fixative is unreported in the selected A09063 caption, and the evidence does not establish whether frozen sections or IF are easier. Assess bronchial autofluorescence when interpreting IF (standard IF practice).
HPA tissue IHC evidence for MAP3K6
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. 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 |
|---|---|---|---|---|
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MAP3K6 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MAP3K6 IHC Tips
Troubleshoot MAP3K6 staining in paraffin sections by checking retrieval, cell type, compartment and controls before comparing chromogenic signal across samples.
What retrieval conditions should I try first for MAP3K6 in paraffin sections?
Use citrate buffer at pH 6.0 for heat induced epitope retrieval at 95–98 °C for 20 min (page retrieval specification). Let sections cool in buffer, then wash consistently before blocking and applying the catalog antibody (standard IHC practice). If staining is weak, compare a modest change in heating time on adjacent sections while keeping antibody incubation and detection constant (standard IHC practice). If tissue lifts or morphology deteriorates, reduce heating intensity and check slide adhesion rather than increasing detection time (standard IHC practice). Record retrieval conditions for every batch so differences in staining remain interpretable (standard IHC practice).
Could fixation explain weak MAP3K6 staining in my sections?
Target specific fixation sensitivity is unknown: the selected product caption describes paraffin embedded human breast carcinoma tissue but does not state its fixative (caption, A09063). Keep fixation method and duration consistent across specimens, and compare sections processed together when assessing weak staining (standard IHC practice). If overfixation is suspected, test adjacent sections with controlled retrieval changes while retaining a morphology check and the same detection settings (standard IHC practice). Poor preservation can obscure cell boundaries and make cytoplasmic scoring unreliable even when chromogen is visible (standard IHC practice). Do not infer a MAP3K6 fixation preference from its tissue staining pattern or protein features (HPA tissue IHC; UniProt O95382).
Should MAP3K6 stain cytoplasm or nuclei in tissue IHC?
Expect widespread cytoplasmic staining as the starting tissue IHC pattern, then assess it within identified cells rather than across the whole section (HPA tissue IHC). High staining is reported in respiratory epithelial cells of bronchus and glandular cells of endometrium, pancreas and stomach (HPA tissue IHC). Cell based IF instead places MAP3K6 mainly in the nucleoplasm, with additional cytosol, plasma membrane and actin filament signal (HPA subcellular). Compare nuclear and cytoplasmic chromogen separately, using the same scoring rules across specimens (standard IHC practice). UniProt supplies no subcellular annotation, so compartment alone cannot establish antibody specificity (UniProt O95382).
Could splice isoforms or phosphorylation change what this antibody detects?
MAP3K6 has three annotated isoforms, and its kinase domain spans residues 648–906 (UniProt O95382). Annotated modified residues include phosphothreonine 806 and phosphoserines 964, 984, 1129 and 1149 (UniProt O95382). The supplied product caption does not map the antibody epitope, so isoform coverage and sensitivity to those modifications cannot be assigned (caption, A09063). If specimens differ in staining, compare adjacent sections under identical retrieval and detection conditions before attributing the difference to isoforms (standard IHC practice). Report the antibody identity and any available epitope information alongside compartment specific scores (standard IHC practice).
How should I investigate MAP3K6 with multiplex immunofluorescence?
For the separate IF/ICC workflow, pair MAP3K6 with a marker that identifies the expected cell type, such as an epithelial marker when examining epithelial cells (HPA tissue IHC; standard IF practice). Choose a fluorophore away from the tissue’s strongest autofluorescence and inspect unstained and single label controls before interpreting overlap (standard IF practice). MAP3K6 has no annotated transmembrane segment, while cell based IF reports nucleoplasmic and cytosolic signal (UniProt O95382; HPA subcellular). Because the catalog epitope is not mapped in the supplied caption, establish permeabilisation empirically when testing access to intracellular epitopes (caption, A09063; standard IF practice). HPA IF images do not validate the catalog antibody for multiplexing (HPA subcellular; caption, A09063).
How can I separate MAP3K6 staining from chromogenic background?
Run a no primary control and inspect it for endogenous peroxidase activity, pigment and nonspecific chromogen deposition before judging weak MAP3K6 signal (standard IHC practice). Include a peroxidase block and keep DAB development time consistent across adjacent sections (standard chromogenic IHC practice). The A09063 tissue image includes a synthesized peptide blocking comparison in paraffin embedded human breast carcinoma, which supports assessing antibody dependent staining under those image conditions (caption, A09063). Peptide blocking alone does not establish the identity of every stained cell or compartment (standard IHC interpretation). Compare suspected signal with intact tissue morphology and the expected broad cytoplasmic tissue pattern (HPA tissue IHC).
What should I measure when comparing MAP3K6 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC reports broad cytoplasmic expression while cell based IF reports prominent nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). For cytoplasmic chromogen, record the percentage of viable target cells at each intensity and calculate an H-score from 0–300 (standard IHC scoring practice). Alternatively, report percentage positive cells or positive cell density per mm² when those measures fit the sampling design (standard IHC scoring practice). Normalise counts to viable cells of the selected type or to sampled tissue area, as appropriate (standard IHC practice). Keep retrieval, DAB development and thresholds constant across compared sections (standard IHC practice).
When is an apparent MAP3K6 positive cell likely to be artefactual?
First confirm that staining follows intact cell boundaries and a reproducible compartment pattern, rather than a section edge or necrotic area (standard IHC interpretation). Cytoplasmic staining across several tissue cell types is plausible, while isolated nuclear signal needs separate review because tissue IHC and cell based IF show different dominant compartments (HPA tissue IHC; HPA subcellular). Check cell identity against morphology and a matching serial section, particularly before calling signal in an unexpected cell population (standard IHC practice). A positive no primary control points toward endogenous enzyme activity or detection background (standard chromogenic IHC practice). Interpret the peptide blocked A09063 image as one specificity control, alongside morphology and independent controls (caption, A09063; standard IHC practice).
Boster reagents
Best MAP3K6 / Mitogen-activated protein kinase kinase kinase 6 IHC Antibodies
A09063 lists IHC and IF applications for human and mouse samples (catalog: applications/reactivity); its IHC image shows a paraffin-embedded human breast carcinoma section (IHC image caption).
Real IHC data
Anti-MEK Kinase-6 MAP3K6 Antibody
Cat # A09063
A09063 will render with an IHC image of paraffin-embedded human breast carcinoma tissue and a peptide-blocked comparison (IHC image caption). The catalog lists IHC and IF applications and human and mouse reactivity for A09063 (catalog: applications/reactivity); no IF image is supplied (catalog: IF images).
Which to pick: For tissue IHC, choose A09063: its own image documents staining in a paraffin-embedded human breast carcinoma section, with fixative unreported (IHC image caption). For IF/ICC, A09063 lists IF, while ICC-specific validation and clonality are unreported (catalog: applications; clone field). For work involving human and mouse samples, A09063 lists reactivity with both species; its IHC image documents human tissue only (catalog: reactivity; IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95382 (M3K6_HUMAN, Mitogen-activated protein kinase kinase kinase 6).
- Human Protein Atlas. MAP3K6 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MAP3K6 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the plasma membrane, cytosol and actin filaments..
- Human Protein Atlas. MAP3K6 antibody validation summary (1 antibodies).
- Salidroside inhibited the proliferation of gastric cancer cells through up-regulating tumor suppressor miR-1343-3p and down-regulating MAP3K6/MMP24 signal molecules. Cancer biology & therapy 2024 — PMC10913707.
- Differential proteomic analysis of lung tissues in rats with high-altitude pulmonary edema and screening of potential biomarkers. Frontiers in physiology 2026 — PMC13616702.
- Role of germline aberrations affecting CTNNA1, MAP3K6 and MYD88 in gastric cancer susceptibility. Journal of medical genetics 2018 — PMC6161648.
- PubMed PMID:17210579 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAP3K6 (Mitogen-activated protein kinase kinase kinase 6). 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: A09063 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Respiratory epithelial 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 breast carcinoma tissue; fixative not specified (datasheet A09063); 2. Fixation: Image fixative and duration unreported (datasheet A09063); 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-MAP3K6, 1:100-1:300 (datasheet A09063); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining–RNA agreement is medium; verify the pattern (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope effects are unestablished (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt O95382, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A09063, PMC10913707, PMC13616702. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAP3K6/immunohistochemistry-ihc-guide.