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MAP2K7 / Dual specificity mitogen-activated protein kinase kinase 7 · IHC design guide
Design Immunohistochemistry for MAP2K7
Plan MAP2K7 chromogenic IHC in paraffin sections using cytoplasmic staining as the tissue pattern (HPA tissue IHC). Use strongly stained spermatogonia as a reference and account for variable staining elsewhere (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAP2K7 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 most tissues (HPA tissue IHC) | |
| Staining pattern | High cytoplasmic staining in spermatogonia; variable elsewhere (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9764) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Caudate+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Skeletal myocytes show no staining despite reported expression (HPA tissue IHC; UniProt) | |
| Regulation | No expression change established (UniProt) | |
| Isoform / epitope | Four isoforms; check whether the epitope is shared (UniProt) |
Section 1
Recommended MAP2K7 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 retrieval (datasheet PB9764). The published MAP2K7 IHC protocol below uses citrate pH 6.0 retrieval (PMC4863692 methods).
| Sample | Paraffin-embedded human testicular cancer tissue; fixative not specified (datasheet PB9764) |
| Fixation | Image fixative and duration unreported (datasheet PB9764); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet PB9764); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9764) |
| Primary antibody | Rabbit anti-MAP2K7, 1-2μg/ml (datasheet PB9764) |
| Primary incubation | Overnight at 4 °C (datasheet PB9764) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9764) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MAP2K7-positive staining in spermatogonia cells of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues at variable levels. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 HIER for the catalog antibody (datasheet PB9764); citrate pH 6.0 is the published alternative (PMC4863692 methods).
Section 2
What Is the Expected MAP2K7 Staining Pattern?
MAP2K7 is a non-transmembrane protein reported in the cytoplasm and nucleus (UniProt O14733 topology and subcellular location). In paraffin-section IHC, expect predominantly cytoplasmic staining across many tissues at variable levels, with high staining in testicular spermatogonia and medium staining in several epithelial and hematopoietic cell populations (HPA: tissue IHC). HPA rates the tissue pattern Supported, with medium consistency between antibody staining and RNA expression (HPA: reliability).
| Cytoplasmic staining in testicular spermatogonia, with weaker or variable staining elsewhere. | This fits the strongest listed tissue observation: spermatogonia are High, while appendix lymphoid tissue and bone-marrow hematopoietic cells are Medium (HPA: tissue IHC). Compare identifiable cells within each section; an entire tissue need not stain uniformly (HPA: cell-level tissue IHC). |
| Signal appears confined to an unexpected compartment, without the expected cytoplasmic pattern. | Check antibody specificity and detection before calling it MAP2K7 (HPA: predominantly cytoplasmic tissue IHC; general IHC practice). Nuclear signal alone is not conclusive evidence of artefact because UniProt also lists the nucleus (UniProt O14733: subcellular location). Membrane-only tissue staining lacks support from the supplied tissue IHC profile (HPA: tissue IHC). |
| Strong staining occurs in cells listed as Not detected, such as oral squamous epithelium. | Consider cross-reactivity or endogenous detection activity and inspect matched controls (HPA: oral mucosa squamous epithelial cells Not detected; general IHC practice). HPA's category describes its observed sample, so a discrepant specimen warrants investigation rather than automatic rejection (HPA: Supported, medium consistency). |
| Diffuse colour covers tissue and spaces between cells, obscuring boundaries. | Treat widespread, poorly localized colour as background until controls support specific staining (general IHC practice). It is difficult to score against HPA's cell-level, predominantly cytoplasmic pattern when the cellular compartment cannot be resolved (HPA: tissue IHC). |
| No signal is visible in an otherwise interpretable testis section. | First verify that spermatogonia are present and assess the detection run with controls (HPA: testis spermatogonia High; general IHC practice). A blank section alone cannot distinguish a technical failure from specimen variation; HPA reports medium consistency between staining and RNA data (HPA: reliability). |
💡Expected MAP2K7 appearanceCall a result consistent with the reference pattern when identifiable testicular spermatogonia show High, predominantly cytoplasmic staining; diffuse colour or strong signal restricted to HPA-listed Not detected cells is suspect and needs controls (HPA: tissue IHC; general IHC practice).
| Choice of tissue and cell-level controls | Testis spermatogonia provide the clearest listed positive comparison; oral squamous epithelial cells are listed as Not detected (HPA: tissue IHC). Score the named cells, because nearby populations can differ (HPA: cell-level tissue IHC). |
| Antibody validation | HPA lists IHC as Supported for HPA001633 and CAB004262; it does not list either as Enhanced in the supplied antibody record (HPA: antibody validation). Supported evidence guides comparison but does not establish specificity in every specimen (general IHC practice). |
| Topology and processing | MAP2K7 has no transmembrane segment or signal peptide in the supplied record (UniProt O14733: topology and processing). The reference tissue pattern is cytoplasmic; a crisp cell-surface-only IHC pattern needs independent validation (HPA: tissue IHC; general IHC practice). |
| Isoforms and phosphorylation | Four isoforms and several modified residues are listed (UniProt O14733: isoforms and modified residues). The supplied record gives no antibody epitope or phospho-specificity, so staining cannot identify an isoform or activation state from this evidence alone (UniProt O14733; HPA: antibody record). |
| Antigen retrieval | Record and optimize retrieval against a positive tissue and a negative control as general paraffin-IHC practice (general IHC practice). The supplied UniProt and HPA evidence does not establish MAP2K7-specific retrieval requirements or fixation sensitivity (UniProt O14733; HPA: tissue IHC). |
| IF/ICC Q&A: should membrane fluorescence match tissue IHC? | HPA's ICC-IF summary lists cytosol and plasma membrane, both uncertain; that observation does not establish a membrane pattern in paraffin tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). Use the separate IF/ICC guide for that application. |
| Situation | Likely cause | Next action |
|---|---|---|
| Testis control is blank, including identifiable spermatogonia. | The run may have a primary-antibody, retrieval, or detection problem; absence of visible spermatogonia can also make the control uninformative (HPA: spermatogonia High; general IHC practice). | Confirm cell identity, then review the run's positive and negative controls and the documented reagent steps before interpreting study sections (general IHC practice). |
| Colour is diffuse across cells, stroma, and empty spaces. | Nonspecific binding, incomplete blocking, or excessive detection signal can obscure a cellular pattern (general IHC practice). HPA describes cytoplasmic staining in identifiable cells (HPA: tissue IHC). | Inspect a no-primary control, check blocking and washes, and adjust antibody or detection conditions using controlled comparisons (general IHC practice). |
| A no-primary control develops chromogenic signal. | Endogenous enzyme activity or nonspecific detection-reagent binding may contribute colour (general IHC practice). Such colour cannot establish MAP2K7 localization (general IHC practice). | Check the detection chemistry and its appropriate endogenous-activity block, then repeat the control before scoring the MAP2K7 slide (general IHC practice). |
| Strong staining appears in oral squamous epithelium or caudate glial cells. | Both populations are listed as Not detected, so cross-reactivity, background, or a specimen-specific difference needs evaluation (HPA: oral mucosa and caudate tissue IHC; general IHC practice). | Compare cell morphology and controls, then seek independent specificity evidence before treating the discrepant signal as MAP2K7 (general IHC practice). |
| Skeletal-muscle myocytes are blank despite UniProt's high-expression statement. | The sources differ: UniProt reports highest tissue expression in skeletal muscle, while HPA lists skeletal-muscle myocytes as Not detected by IHC (UniProt O14733: tissue specificity; HPA: tissue IHC). | Do not use myocyte staining alone to accept or reject the run; assess an HPA-listed positive cell population and the run controls (HPA: testis spermatogonia High; general IHC practice). |
| Signal is exclusively nuclear or sharply restricted to cell borders. | UniProt permits nuclear localization, but HPA's tissue IHC profile is predominantly cytoplasmic; ICC-IF membrane localization is uncertain (UniProt O14733: subcellular location; HPA: tissue IHC and ICC-IF). | Review morphology, compartment resolution, and controls; validate an atypical pattern independently before reporting it as the expected tissue IHC result (general IHC practice). |
Experiment evidence
IHC & IF images showing MAP2K7 staining
PB9764 · IHCIHC analysis of MEK7/MAP2K7 using anti-MEK7/MAP2K7 antibody (PB9764). MEK7/MAP2K7 was detected in a paraffin-embedded section of human testi
PB9764 · IFIF analysis of MEK7/MAP2K7 using anti-MEK7/MAP2K7 antibody (PB9764). MEK7/MAP2K7 was detected in an immunocytochemical section of PC-3 cells
A02713S271-1 · IHCImmunohistochemistry analysis of paraffin-embedded human brain, using MAP2K7 (Phospho-Ser271) Antibody. The picture on the right is blocked
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000076984-MAP2K7/tissue/cerebral+cortex#imid_1386388
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000076984-MAP2K7/tissue/colon#imid_1386298
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000076984-MAP2K7/tissue/liver#imid_1386307
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000076984-MAP2K7/tissue/kidney#imid_1386316
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA064711; green = MAP2K7, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA064711; green = MAP2K7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAP2K7 IHC & IF
🧪Run testis first; spermatogonia should stain (HPA: High in testis spermatogonia cells). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected); on the testis slide, neighboring cells without specific DAB staining can serve as internal negatives, although the supplied HPA rows do not identify a negative testicular cell type.
Positive control tissue: Testis (Spermatogonia cells, HPA High)
Negative control tissue: Caudate (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAP2K7 in HEK293, PC-3, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Plasma membrane (uncertain), Cytosol (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, matched rabbit IgG isotype control, and MAP2K7 knockout or validated peptide-block control (caption: rabbit primary antibody; standard IHC practice). For the testis section, check endogenous peroxidase and biotin background because the reported detection uses a biotin-based complex and DAB (selected-SKU caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact selected-SKU caption does not state the fixative (selected-SKU caption: fixative unreported). The reported paraffin-section staining used heat-mediated retrieval in EDTA pH 8.0, but the evidence does not establish whether retrieval is essential (selected-SKU caption: EDTA retrieval). Neither frozen sections nor IF are established as easier here; for the reported testis DAB method, endogenous biotin or peroxidase may produce background that the controls should reveal (selected-SKU caption: SABC and DAB; standard IHC practice).
HPA tissue IHC evidence for MAP2K7
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 |
|---|---|---|---|---|
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced MAP2K7 IHC Tips
Troubleshoot MAP2K7 staining by anchoring the chromogenic IHC workflow to the PB9764 tissue image and checking compartment, cell type, and controls.
Which retrieval condition should I start with for MAP2K7 in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet PB9764). The catalog antibody produced a tissue image after this retrieval, followed by 10% goat serum blocking and 2 μg/ml primary antibody overnight at 4°C (datasheet PB9764). If staining is weak, compare retrieval duration on adjacent sections while keeping detection and exposure to DAB consistent (standard IHC practice). Include a no-primary control and tissue with an expected positive cell population, such as testicular spermatogonia (HPA: High in spermatogonia cells), before treating stronger DAB signal as better retrieval. Excessive heating can damage morphology and make compartment scoring unreliable (standard IHC practice).
Could fixation explain weak or uneven MAP2K7 staining?
The PB9764 tissue caption identifies a paraffin section but does not state its fixative, so MAP2K7 sensitivity to fixation is unknown for this antibody (datasheet PB9764). Record the fixative, fixation interval, processing schedule, and section age for each specimen before comparing staining intensity (standard IHC practice). Compare adjacent sections processed together using the stated EDTA pH 8.0 retrieval and 2 μg/ml primary concentration (datasheet PB9764). If weak staining tracks a processing batch, repeat with a matched positive control rather than attributing the change to MAP2K7 biology (standard IHC practice). Preserve morphology when adjusting retrieval, because damaged cells cannot support dependable compartment or cell-type scoring (standard IHC practice).
Should MAP2K7 appear in nuclei, cytoplasm, or at the cell surface?
Prioritise interpretable cytoplasmic staining in tissue: HPA reports cytoplasmic expression in most tissues at variable levels (HPA tissue IHC). UniProt lists both nucleus and cytoplasm, while HPA subcellular data place MAP2K7 in cytosol and at an uncertain plasma membrane location (UniProt O14733 subcellular; HPA subcellular). MAP2K7 has no transmembrane segment, so a sharp membrane-only DAB rim needs independent validation before assignment to this kinase (UniProt O14733 topology). Compare suspect staining with a no-primary control and inspect the counterstain for intact cell borders and nuclei (standard IHC practice). Score nuclear and cytoplasmic compartments separately when both are convincing, using consistent thresholds across specimens (standard IHC practice).
Can this stain distinguish MAP2K7 isoforms or phosphorylation states?
Do not assign PB9764 staining to a particular splice form: MAP2K7 has four listed isoforms, but the supplied tissue caption does not map this antibody’s epitope (UniProt O14733 isoforms; datasheet PB9764). The kinase domain spans residues 120–380, and reported modifications include phosphorylation at 271, 275, and 411 (UniProt O14733 domains and modified residues). Without epitope mapping or phospho-specific validation, DAB intensity cannot distinguish total protein from a modified form (standard IHC interpretation). Compare staining across similarly processed sections and use an independently validated reagent if isoform or phosphorylation specificity is essential (standard IHC practice). Document the antibody and retrieval condition alongside every resulting score (standard IHC practice).
How should I adapt the MAP2K7 localisation check to multiplex IF?
Treat IF as a separately optimised application; the supplied PB9764 image documents chromogenic staining in a paraffin section (datasheet PB9764). Pair MAP2K7 with a validated marker for the expected cell type, such as a spermatogonia marker when examining testis, and include single-label controls to check channel separation (HPA: High in spermatogonia cells; standard IF practice). Choose fluorophores after inspecting unstained tissue autofluorescence, placing the weaker signal in a cleaner spectral channel (standard IF practice). MAP2K7 lacks a transmembrane segment and is reported in cytoplasm and nucleus, so test permeabilisation for intracellular access rather than assuming a surface-facing epitope (UniProt O14733 topology and subcellular). Compare compartment patterns with no-primary controls before interpreting colocalisation (standard IF practice).
How can I separate diffuse MAP2K7 signal from chromogenic background?
Compare the stained section with a no-primary control to identify background from secondary detection, endogenous enzymes, or tissue pigment (standard IHC practice). The PB9764 image used a biotinylated secondary, a streptavidin–biotin complex, and DAB, so check the detection system when diffuse brown signal persists without primary antibody (datasheet PB9764). Include an appropriate peroxidase block and keep DAB development consistent between test and control sections (standard chromogenic IHC practice). The reported workflow used 10% goat serum blocking and 2 μg/ml primary overnight at 4°C; change one variable at a time if background remains high (datasheet PB9764; standard IHC practice). Interpret faint diffuse staining cautiously because tissue expression varies (HPA tissue IHC).
What is a defensible way to quantify MAP2K7 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then apply the same threshold and DAB development conditions to every section (standard IHC practice). An H-score combines the percentage of cells at each intensity; alternatively, report % positive cells or positive-cell density per mm² within a prespecified region (standard IHC quantification). Normalise counts to the number of eligible cells or measured viable tissue area, excluding folds, necrosis, and section edges (standard IHC practice). Record cytoplasmic and nuclear scores separately because tissue IHC is predominantly cytoplasmic while UniProt also lists nuclear localisation (HPA tissue IHC; UniProt O14733 subcellular). Use matched controls and blinded scoring when comparing groups (standard IHC practice).
When is a MAP2K7-positive cell credible rather than an artefact?
Look for reproducible staining in intact cells with the expected compartment pattern: tissue IHC is predominantly cytoplasmic, while nuclear localisation is also listed (HPA tissue IHC; UniProt O14733 subcellular). Testicular spermatogonia are a useful positive reference, whereas HPA reports no detection in skeletal-muscle myocytes despite UniProt describing high skeletal-muscle expression overall (HPA tissue IHC; UniProt O14733 tissue specificity). That discrepancy calls for cell-resolved review rather than an assumption that every myocyte must stain. Reject isolated staining at section edges, in necrotic areas, or in a no-primary control as evidence of specific MAP2K7 expression (standard IHC practice). If brown signal persists without primary antibody, investigate endogenous enzyme activity and the biotin-based detection workflow (standard IHC practice; datasheet PB9764).
Boster reagents
Best MAP2K7 / Dual specificity mitogen-activated protein kinase kinase 7 IHC Antibodies
The catalog includes IHC images from human paraffin-embedded testicular cancer tissue and brain, plus IF data from PC-3 cells (PB9764 and A02713S271-1 image captions).
Real IHC data
Anti-MEK7/MAP2K7 Antibody ®
Cat # PB9764
Real IHC data
Anti-Phospho-MEK-7 (S271) MAP2K7 Antibody
Cat # A02713S271-1
PB9764 has IHC data from a human paraffin-embedded testicular cancer section and IF data from PC-3 cells (PB9764 image captions). A02713S271-1 has phospho-Ser271 IHC data from paraffin-embedded human brain, including a phospho-peptide blocking image (A02713S271-1 IHC image caption).
Which to pick: Choose PB9764 for tissue IHC when a documented paraffin-section workflow is useful; its caption reports heat retrieval in EDTA at pH 8.0, while the fixative is unreported (PB9764 IHC image caption). PB9764 also has documented IF staining in PC-3 cells, whereas A02713S271-1 is the choice for phospho-Ser271 IHC in human brain sections (PB9764 IF image caption; A02713S271-1 IHC image caption). PB9764 lists the broadest species reactivity—human, monkey, mouse and rat—but its supplied IHC image documents human tissue only (PB9764 catalog reactivity; PB9764 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14733 (MP2K7_HUMAN, Dual specificity mitogen-activated protein kinase kinase 7).
- Human Protein Atlas. MAP2K7 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MAP2K7 subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. MAP2K7 antibody validation summary (3 antibodies).
- miR-125b inhibited epithelial-mesenchymal transition of triple-negative breast cancer by targeting MAP2K7. OncoTargets and therapy 2016 — PMC4863692.
- LncRNA PCED1B-AS1 mediates miR-3681-3p/MAP2K7 axis to promote metastasis, invasion and EMT in gastric cancer. Biology direct 2024 — PMC11064384.
- Inhibition of the MAP2K7-JNK pathway with 5Z-7-oxozeaenol induces apoptosis in T-cell acute lymphoblastic leukemia. Oncotarget 2021 — PMC8416565.
- PubMed PMID:9372971 — UniProt-cited evidence.
- PubMed PMID:9312068 — UniProt-cited evidence.
- PubMed PMID:9535930 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAP2K7 (Dual specificity mitogen-activated protein kinase kinase 7). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9764). Validated IHC antibody: PB9764 at 1-2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Spermatogonia cells); negative Caudate. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human testicular cancer tissue; fixative not specified (datasheet PB9764); 2. Fixation: Image fixative and duration unreported (datasheet PB9764); 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: EDTA pH 8.0 (datasheet PB9764); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9764); 8. Primary antibody: Rabbit anti-MAP2K7, 1-2μg/ml (datasheet PB9764); 9. Primary incubation: Overnight at 4 °C (datasheet PB9764). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Skeletal myocytes show no staining despite reported expression (HPA tissue IHC; UniProt); Isoform / epitope: Four isoforms; check whether the epitope is shared (UniProt); Regulation: No expression change established (UniProt). Evidence: UniProt O14733, HPA tissue IHC · Supported, HPA ICC-IF · plasma membrane and cytosol., Datasheet PB9764, PMC4863692. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAP2K7/immunohistochemistry-ihc-guide.