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MAP2K1 / Dual specificity mitogen-activated protein kinase kinase 1 · IHC design guide
Design Immunohistochemistry for MAP2K1
Plan chromogenic IHC for MAP2K1 in paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A00292). Compare cytoplasmic staining with the tissue profile and account for its low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAP2K1 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic in glandular cells and lung macrophages (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00292) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Breast+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). Selected-image fixative and duration unreported (datasheet A00292); verify before use. | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation is not specified (UniProt) | |
| Isoform / epitope | Two isoforms; map the antibody epitope across both (UniProt) |
Section 1
Recommended MAP2K1 IHC & IF Protocols
The catalog antibody uses EDTA pH 8.0 heat retrieval (datasheet A00292). Published IHC protocols cover HSCC tissue (PMC11984367) and liver sections (PMC7641625).
| Sample | Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A00292) |
| Fixation | Image fixative and duration unreported (datasheet A00292); 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 A00292); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00292) |
| Primary antibody | Rabbit anti-MAP2K1, 2-5 μg/ml (datasheet A00292) |
| Primary incubation | Overnight at 4 °C (datasheet A00292) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A00292) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MAP2K1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet A00292); assess the published conditions separately because neither article specifies a retrieval buffer (PMC11984367; PMC7641625).
Section 2
What Is the Expected MAP2K1 Staining Pattern?
MAP2K1 staining in paraffin sections should be predominantly cytoplasmic, especially in glandular cells and lung macrophages reported at medium intensity by HPA (HPA tissue IHC: general cytoplasmic expression; medium in listed cells). Its lack of a transmembrane segment is consistent with an intracellular protein, although UniProt also lists nuclear and cell-cycle-associated locations (UniProt Q02750: topology and subcellular location). HPA rates its tissue IHC as Approved but reports low consistency between staining and RNA data (HPA tissue IHC: reliability).
| Cytoplasmic chromogen in adrenal or appendix glandular cells, with discernible cell outlines. | This fits the reported medium staining in those cell types (HPA tissue IHC: adrenal gland and appendix). Score the relevant cells, since HPA reports levels by cell type; the result alone does not prove antibody specificity (HPA tissue IHC: cell-level observations and low staining–RNA consistency). |
| Strong staining confined to luminal material or extracellular spaces, with little cellular staining. | This does not match the reported general cytoplasmic IHC profile or a protein without a signal peptide or transmembrane segment (HPA tissue IHC: profile; UniProt Q02750: processing and topology). Consider deposited chromogen or nonspecific staining (general IHC practice). Nuclear staining alone needs context: UniProt also lists the nucleus (UniProt Q02750: subcellular location). |
| Respiratory epithelial cells stain strongly while nearby lung macrophages do not. | HPA reports respiratory epithelial cells in bronchus as not detected and lung macrophages as medium (HPA tissue IHC: bronchus and lung). The mismatch raises possible cross-reactivity or endogenous detection activity; assess cell identity and controls before assigning MAP2K1 expression (general IHC practice). |
| Chromogen covers most of the section, including spaces between cells. | A diffuse deposit obscures the cell-level cytoplasmic pattern used for interpretation (HPA tissue IHC: general cytoplasmic expression). Excess detection reagent, incomplete blocking or inadequate washing can produce background (general IHC practice); background alone cannot establish MAP2K1 distribution. |
| No cellular signal in an adrenal gland or appendix section selected as a positive reference. | HPA reports medium staining in glandular cells at these sites, so absence warrants a run-level check (HPA tissue IHC: adrenal gland and appendix). Confirm that the expected cells are present, then review antibody use and chromogenic detection with controls (general IHC practice). |
💡Expected MAP2K1 appearanceCall a positive result when identifiable adrenal or appendix glandular cells show predominantly cytoplasmic staining near HPA's reported medium level; extracellular deposit or indiscriminate staining is suspect (HPA tissue IHC: profile and cell levels; UniProt Q02750: topology; general IHC practice).
| Cell type and tissue selection | HPA reports medium staining in adrenal, appendix, duodenum and gallbladder glandular cells and lung macrophages, but not detected in breast adipocytes or bronchial respiratory epithelium (HPA tissue IHC). Use the named cell population when comparing sections; a whole tissue is not uniformly positive or negative (HPA tissue IHC: cell-level observations). |
| Compartment and topology | General cytoplasmic IHC staining is reported by HPA; UniProt lists cytoplasm, nucleus and locations at mitotic structures, with no signal peptide or transmembrane segment (HPA tissue IHC: profile; UniProt Q02750: subcellular location, processing and topology). Interpret a membrane-like edge with cellular context; these records do not set an IHC membrane-intensity threshold. |
| Strength of tissue evidence | The HPA tissue IHC status is Approved, with low consistency between antibody staining and RNA expression data (HPA tissue IHC: reliability and description). Three listed antibodies have Approved IHC status, but the supplied record gives no target-specific fixation or retrieval sensitivity (HPA antibodies: IHC status; supplied UniProt and HPA records). |
| Isoforms and modified residues | UniProt lists 2 isoforms and phosphorylation sites including Ser218 and Ser222 (UniProt Q02750: isoforms and modified residues). Without a supplied epitope or phospho-specific antibody record, these facts cannot predict differential chromogenic staining or establish that a dark cell has activated MAP2K1 (UniProt Q02750: record scope). |
| IF/ICC Q: What location should be expected? | A: HPA reports supported plasma membrane and cytosol localization, plus basal body localization, in ICC-IF (HPA subcellular: main and additional locations). Those fluorescence observations provide context, but do not replace the general cytoplasmic pattern reported for tissue IHC (HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected glandular cells are unstained in a positive reference section. | The cells may be absent from the field, or the staining run may have failed; HPA reports medium staining for adrenal and appendix glandular cells (HPA tissue IHC). | First locate the specified cell type. Then check the catalog antibody's IHC-P instructions, retrieval step, antibody dilution and detection controls as general IHC workflow checks; no MAP2K1-specific retrieval effect is supplied (general IHC practice; supplied record scope). |
| Strong nuclear signal appears without convincing cytoplasmic staining. | UniProt includes nuclear localization, so a nuclear component is possible, but HPA describes general cytoplasmic tissue staining (UniProt Q02750: subcellular location; HPA tissue IHC: profile). | Compare the same cell type with a reference section and negative detection control. Treat a nucleus-only result as unresolved until the cellular pattern and controls support it (general IHC practice). |
| Bronchial epithelium stains strongly while macrophage staining is weak. | The relative pattern conflicts with HPA's not-detected bronchial respiratory epithelium and medium lung macrophage observations (HPA tissue IHC: bronchus and lung). Nonspecific antibody binding or endogenous chromogenic activity is possible (general IHC practice). | Verify the cell types morphologically, then examine a control omitting primary antibody and review blocking and detection steps (general IHC practice). Do not treat one discordant field as a new tissue-wide expression profile. |
| Most cells and extracellular areas show uniform brown haze. | Diffuse background prevents assessment of the reported cytoplasmic cell pattern (HPA tissue IHC: profile). Excess primary or detection reagent and insufficient washing are common general IHC causes (general IHC practice). | Inspect the negative detection control; optimize reagent concentration, blocking and washes according to the assay workflow (general IHC practice). Reassess only when individual cells and compartments are distinguishable. |
| A whole section is called negative because one cell population lacks staining. | HPA levels apply to named cell populations: breast adipocytes and bronchial respiratory epithelial cells are not detected, while several listed glandular populations and lung macrophages are medium (HPA tissue IHC). | Record tissue, cell type, compartment and intensity separately. Compare with an HPA observation for that specific population before changing the section-level interpretation (HPA tissue IHC: cell-level observations). |
| A dark cytoplasmic result is interpreted as activated MAP2K1. | Routine total-protein IHC intensity does not identify a phosphorylation state; UniProt lists modified residues but the supplied records do not define a phospho-specific staining reagent (UniProt Q02750: modified residues; supplied record scope). | Report the observed compartment and intensity without an activation claim. A phosphorylation conclusion requires a separately validated phospho-specific assay and its controls (general IHC practice). |
Experiment evidence
IHC & IF images showing MAP2K1 staining
A00292 · IHCIHC analysis of MEK1/MAP2K1 using anti-MEK1/MAP2K1 antibody (A00292). MEK1/MAP2K1 was detected in a paraffin-embedded section of human ovari
A00292 · IFIF analysis of MEK1/MAP2K1 using anti-MEK1/MAP2K1 antibody (A00292). MEK1/MAP2K1 was detected in an immunocytochemical section of TPC1 cells
M00292-5 · IHCHuman kidney was stained with anti-MEK1 rabbit antibody
M00292-5 · IHCHuman lung was stained with anti-MEK1 rabbit antibody
M00292-1 · IHCImmunohistochemical analysis of paraffin-embedded human kidney, using MEK1/2 Antibody.
M00292-1 · IFImmunofluorescent analysis of Hela cells, using MEK1/2 Antibody .
M00292-2 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000169032-MAP2K1/tissue/cerebral+cortex#imid_6822488
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000169032-MAP2K1/tissue/colon#imid_6822414
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000169032-MAP2K1/tissue/liver#imid_6822407
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000169032-MAP2K1/tissue/kidney#imid_6822416
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026430; green = MAP2K1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA026430; green = MAP2K1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAP2K1 IHC & IF
🧪Run adrenal gland first and assess staining in its glandular cells (HPA: Medium in adrenal gland glandular cells). Use breast adipocytes as the negative tissue (HPA: Not detected in breast adipocytes); on the adrenal slide, treat adjacent unstained cells as internal negatives only after confirming they remain at background, since HPA provides no negative cell type for that tissue.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Breast (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAP2K1 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), KOLF2.1J, with annotated localisation: Plasma membrane (supported), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), a host- and clonality-matched isotype control, and MAP2K1-knockout tissue or cells as a biological specificity control (selected A00292 caption: rabbit primary and peroxidase-conjugated secondary; standard IHC practice). Quench endogenous peroxidase and check background before interpreting adrenal staining with HRP/DAB (selected A00292 caption: HRP/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A00292 paraffin-section caption does not state a fixative (selected A00292 caption: fixative unreported). That caption uses heat-mediated EDTA retrieval at pH 8.0, but whether retrieval is required or another method works is unreported (selected A00292 caption: EDTA retrieval). No matched frozen-section or IF protocol establishes either as easier; adrenal HRP/DAB staining requires an endogenous-peroxidase background check (selected A00292 caption: HRP/DAB detection; standard IHC practice).
HPA tissue IHC evidence for MAP2K1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAP2K1 IHC Tips
Troubleshoot MAP2K1 staining in paraffin sections by checking retrieval, controls, cellular location, and scoring before interpreting pathway activity.
What retrieval conditions should I try first for MAP2K1 in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A00292). The documented ovarian cancer tissue image used this retrieval before overnight incubation with 2 μg/ml antibody at 4°C (datasheet A00292). If staining is weak, vary heating and cooling duration on matched sections while keeping antibody concentration and detection conditions constant (standard IHC practice). Compare signal in intact cells with a no-primary control, because MAP2K1 is reported in the cytoplasm and at the membrane, and stronger overall DAB staining alone does not establish specific retrieval (UniProt Q02750 subcellular; HPA subcellular; standard IHC practice).
Could fixation explain weak MAP2K1 staining in my sections?
Target-specific MAP2K1 sensitivity to fixation is unknown from the supplied evidence; the selected image identifies paraffin embedding but does not state its fixative (datasheet A00292). Record the fixative, fixation duration, and processing history for each specimen, then compare matched sections using the same EDTA pH 8.0 retrieval and detection run (datasheet A00292; standard IHC practice). Include a no-primary control and a previously stained reference section to distinguish a specimen problem from run-wide loss of signal (standard IHC practice). Do not infer fixation tolerance from the reported cytoplasmic staining pattern or MAP2K1 modifications; neither measures fixation sensitivity (HPA tissue IHC; UniProt Q02750 modified residues).
Where should MAP2K1 staining appear, and when is localisation suspicious?
Expect predominantly cytoplasmic staining in tissue sections, while allowing membrane-associated signal where cell boundaries are resolvable (HPA tissue IHC: general cytoplasmic expression; HPA subcellular: plasma membrane and cytosol). MAP2K1 has no transmembrane segment, and its membrane localisation is reported as probably regulated by interaction with KSR1 (UniProt Q02750 topology and subcellular). Nuclear and mitotic structures are also reported, including centrosomes during prometaphase, the midzone during anaphase, and the midbody during telophase (UniProt Q02750 subcellular). If DAB appears exclusively nuclear or confined to damaged section edges, check morphology and controls before assigning it to MAP2K1 (standard IHC practice).
Could isoforms or phosphorylation change what this antibody detects?
MAP2K1 has 2 listed isoforms and a protein kinase domain spanning residues 68–361 (UniProt Q02750 isoforms and domains). The supplied caption does not identify the antibody epitope, so staining cannot be assigned to one isoform or assumed to recognise both (datasheet A00292; UniProt Q02750 isoforms). MAP2K1 has reported phosphorylation sites including Ser218 and Ser222, but their effect on this antibody's binding is unreported (UniProt Q02750 modified residues; datasheet A00292). If isoform or phosphorylation state matters, obtain epitope validation and compare with an independently validated assay on matched material before interpreting staining differences (standard IHC practice).
How should I check MAP2K1 localisation by multiplex immunofluorescence?
On a separate IF/ICC workflow, multiplex MAP2K1 with a validated marker for the cell population being evaluated, such as macrophages when examining the reported lung macrophage signal (HPA tissue IHC: lung macrophages, Medium; standard IF practice). Select a fluorophore channel after checking tissue autofluorescence and single-colour controls; a longer-wavelength channel can improve separation when shorter-wavelength autofluorescence is strong (standard IF practice). Permeabilise sufficiently to reach cytosolic MAP2K1 and the cytosolic side of membrane-associated MAP2K1, then check that cell boundaries remain interpretable (UniProt Q02750 topology; HPA subcellular; standard IF practice). Keep IF exposure and segmentation settings consistent across compared specimens (standard IF practice).
How can I reduce diffuse DAB background without losing MAP2K1 signal?
First inspect a no-primary control and block endogenous peroxidase before HRP and DAB detection, since either background source can mimic weak cytoplasmic staining (standard IHC practice). The documented section used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (datasheet A00292). If background persists, titrate the primary antibody and optimise washing or blocking on adjacent sections while retaining the same retrieval condition (standard IHC practice; datasheet A00292). Judge improvements by preserved cellular detail and reduced no-primary signal, rather than by darker MAP2K1 staining alone (standard IHC practice).
How should I score MAP2K1 IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; MAP2K1 tissue staining is described as generally cytoplasmic (HPA tissue IHC). An H-score combines percentages at intensity grades 0–3 into a 0–300 score, while percent-positive cells provides a simpler measure when intensity is unreliable (standard IHC practice). Normalise counts to the number of evaluable cells in the chosen population, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC practice). Apply one threshold, counterstain assessment, and exclusion rule for damaged or necrotic regions across the whole comparison (standard IHC practice).
How do I distinguish true MAP2K1 staining from artefact or pathway activation?
A credible result places staining in intact cells with predominantly cytoplasmic distribution, with possible membrane-associated signal, and exceeds the no-primary control (HPA tissue IHC; HPA subcellular; standard IHC practice). Check cell identity: lung macrophages have reported medium staining, whereas bronchial respiratory epithelial cells are reported as not detected in the sampled profiles (HPA tissue IHC). Discount staining restricted to section edges, necrotic areas, or sites positive in the no-primary control, and investigate endogenous peroxidase when using HRP and DAB (standard IHC practice). Total MAP2K1 staining does not by itself establish phosphorylation or MAPK/ERK pathway activity; those require a separately validated activity readout (UniProt Q02750 function and modified residues; standard IHC interpretation).
Boster reagents
Best MAP2K1 / Dual specificity mitogen-activated protein kinase kinase 1 IHC Antibodies
These anti-MAP2K1 catalog entries include human tissue IHC images and cell IF images (catalog image captions); all four list human, mouse, and rat reactivity (catalog reactivity fields).
Real IHC data
Anti-MEK1/MAP2K1 Antibody ®
Cat # A00292
Real IHC data
Anti-MEK1 Rabbit Monoclonal Antibody
Cat # M00292-5
Real IHC data
Anti-MEK1/2 MAP2K1 Rabbit Monoclonal Antibody
Cat # M00292-1
Real IF data
Anti-MEK1 MAP2K1 Rabbit Monoclonal Antibody
Cat # M00292-2
A00292 shows IHC in a human ovarian cancer paraffin section and IF in TPC1 and U2OS cells (A00292 image captions); M00292-5 shows IHC in human kidney and lung (M00292-5 image captions). M00292-1 shows IHC in a human kidney paraffin section and lists IF/ICC (M00292-1 image caption; catalog applications), while M00292-2 shows IF at 1:50 and lists IHC without an IHC image (M00292-2 IF caption; catalog applications and image captions).
Which to pick: Choose A00292 for paraffin tissue IHC when a documented starting method helps: its human ovarian cancer caption specifies EDTA retrieval at pH 8.0 and 2 μg/ml antibody; the fixative is unreported (A00292 IHC caption). For IF/ICC, A00292 has cell images at 5 μg/ml, while M00292-2 is a monoclonal option with an IF image at 1:50 (A00292 IF captions; M00292-2 catalog description and IF caption). For cross-species planning, all four list human, mouse, and rat reactivity, but their supplied tissue IHC images are human (catalog reactivity fields; IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q02750 (MP2K1_HUMAN, Dual specificity mitogen-activated protein kinase kinase 1).
- Human Protein Atlas. MAP2K1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MAP2K1 subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the basal body..
- Human Protein Atlas. MAP2K1 antibody validation summary (3 antibodies).
- IGF-1 Induces GHRH Neuronal Axon Elongation during Early Postnatal Life in Mice. PloS one 2017 — PMC5226784.
- Inhibition of the long non-coding RNA MALAT1 downregulates MAP2K1, suppressing the progression of hypopharyngeal squamous cell carcinoma. Biomolecules & biomedicine 2025 — PMC11984367.
- Sclerosing melanocytic tumors with MAP2K1 in frame deletions and 15q gains: A distinctive pathway of nevogenesis with reproducible morphology. Virchows Archiv : an international journal of pathology 2026 — PMC13368881.
- Mechanism of Paeoniflorin in the Treatment of Bile Duct Ligation-Induced Cholestatic Liver Injury Using Integrated Metabolomics and Network Pharmacology. Frontiers in pharmacology 2020 — PMC7641625.
- PubMed PMID:1281467 — UniProt-cited evidence.
- PubMed PMID:8388392 — UniProt-cited evidence.
- PubMed PMID:10409742 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAP2K1 (Dual specificity mitogen-activated protein kinase kinase 1). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00292). Validated IHC antibody: A00292 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Breast. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovarian cancer tissue; fixative not specified (datasheet A00292); 2. Fixation: Image fixative and duration unreported (datasheet A00292); 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 A00292); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00292); 8. Primary antibody: Rabbit anti-MAP2K1, 2-5 μg/ml (datasheet A00292); 9. Primary incubation: Overnight at 4 °C (datasheet A00292). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A00292); verify before use.; Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: Two isoforms; map the antibody epitope across both (UniProt); Regulation: Expression regulation is not specified (UniProt). Evidence: UniProt Q02750, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A00292, PMC11984367, PMC7641625. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAP2K1/immunohistochemistry-ihc-guide.