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MAP2K1 / Dual specificity mitogen-activated protein kinase kinase 1 · IHC design guide
Design Immunohistochemistry for MAP2K1
Plan MAP2K1 paraffin IHC around general cytoplasmic staining (HPA tissue IHC). Lung macrophages show medium staining, while breast adipocytes are undetected; interpret these references in light of the low consistency between staining and 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 staining across multiple cell types (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 paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 2 isoforms; full-length chain, epitope coverage unknown (UniProt) |
Section 1
Recommended MAP2K1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published MAP2K1 staining protocols for hypopharyngeal squamous cell carcinoma and liver sections (PMC11984367; 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 heat-mediated EDTA retrieval at pH 8.0 (datasheet A00292); the published excerpts do not specify retrieval conditions (PMC11984367; PMC7641625).
Section 2
What Is the Expected MAP2K1 Staining Pattern?
MAP2K1 staining in paraffin sections is expected mainly in the cytoplasm: HPA reports general cytoplasmic expression, including medium staining in duodenal glandular cells and lung macrophages (HPA tissue IHC). UniProt also lists nuclear, membrane and mitotic structures, and reports no transmembrane segment (UniProt Q02750). Interpret tissue comparisons cautiously: HPA rates its IHC data Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in duodenal glandular cells or lung macrophages. | This fits the reported IHC pattern and medium staining level in those specific cell populations (HPA tissue IHC). Judge the cells on the slide; the HPA entries do not imply that every cell in either tissue should stain. |
| Strong staining confined to nuclei, with little cytoplasmic signal. | Recheck the pattern against HPA's general cytoplasmic IHC profile (HPA tissue IHC). UniProt also lists nuclear localization (UniProt Q02750), so nuclear signal alone does not prove an artefact; compare it with a positive tissue and controls. |
| Signal mainly in a cell population reported as undetected. | For example, HPA reports no detection in bronchial respiratory epithelial cells (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic activity, then examine a no-primary control. An HPA negative entry is a comparison point, not a universal absence claim. |
| Diffuse colour across tissue, stroma and blank areas. | An indiscriminate deposit does not match HPA's cell-resolved cytoplasmic pattern (HPA tissue IHC). In general IHC practice, assess background from detection reagents, blocking and washing with a no-primary control before scoring MAP2K1-positive cells. |
| No staining in a selected HPA medium-staining cell population. | Absence in duodenal glandular cells or lung macrophages conflicts with the cited HPA observations (HPA tissue IHC), but does not establish biological loss. First check tissue preservation, antibody and detection performance, and whether those cells are present in the section. |
💡Expected MAP2K1 appearanceCall a result positive when identifiable duodenal glandular cells or lung macrophages show discernible cytoplasmic chromogen at a level consistent with HPA's medium category; diffuse deposit outside cells is suspect (HPA tissue IHC).
| Tissue and cell selection | HPA reports medium staining in several glandular and neuronal populations and lung macrophages, but no detection in selected epithelial and other cells (HPA tissue IHC). Select and score the named cell population rather than treating an entire organ as uniformly positive. |
| Compartment and topology | General cytoplasmic IHC staining is the tissue reference (HPA tissue IHC). UniProt also records nucleus, membrane and mitotic structures, with no transmembrane segment (UniProt Q02750); these annotations do not predict that every location will resolve in chromogenic sections. |
| Antibody interpretation | Three listed antibodies have Approved IHC status; HPA reports low consistency between staining and RNA expression for this tissue profile (HPA antibody validation; HPA tissue IHC). Treat an unexpected distribution as a result to validate, not a definitive expression map. |
| IF/ICC Q: Should the section resemble an ICC image? | A: HPA supports plasma membrane and cytosol localization, plus basal body localization, in ICC-IF (HPA subcellular). Use that as compartment context; assess paraffin-section staining against HPA tissue IHC. IF/ICC methods belong in the separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| A selected positive population has no visible chromogen. | The chosen cells may be absent, or the staining run may have failed; HPA reports medium staining in duodenal glandular cells and lung macrophages (HPA tissue IHC). | Verify cell identity on the counterstained section and inspect a same-run positive control. As general IHC practice, check antibody, retrieval and detection steps against the catalog antibody's IHC-P instructions. |
| Colour appears throughout the slide, including regions without target cells. | Noncellular background can obscure HPA's general cytoplasmic pattern (HPA tissue IHC). In general IHC practice, detection reagent background or insufficient washing are possible causes. | Review a no-primary control and the detection blank; adjust blocking, washing or chromogen development using standard IHC controls before interpreting faint cellular staining. |
| Macrophages show unusually intense chromogen or a no-primary control stains. | Endogenous detection activity is a possible source of chromogenic signal in general IHC practice. HPA reports medium MAP2K1 staining in lung macrophages (HPA tissue IHC), so cell identity alone cannot resolve the signal's origin. | Compare the no-primary control on the same tissue and check the detection system's endogenous-activity blocking step. Score MAP2K1 only where specific cellular staining exceeds that control. |
| Nuclear signal dominates while cytoplasmic staining is weak. | This departs from the general tissue IHC profile (HPA tissue IHC), although nuclear localization is listed by UniProt (UniProt Q02750). | Compare a known-positive section and no-primary control under the same scoring criteria. Report the compartment observed; avoid calling nuclear staining either specific or artefactual from location alone. |
| Cells reported as undetected show clear staining. | HPA reports no detection in bronchial respiratory epithelial cells and breast adipocytes (HPA tissue IHC). Unexpected signal could reflect background or antibody cross-reactivity; the HPA profile has low RNA–staining consistency (HPA tissue IHC). | Confirm the cell type and examine no-primary and positive controls. If the signal persists, document the discrepancy and seek independent antibody or orthogonal evidence before assigning MAP2K1 expression. |
| ICC-IF membrane signal seems inconsistent with cytoplasmic tissue IHC. | HPA supports plasma membrane and cytosol localization in ICC-IF, while its tissue IHC profile is generally cytoplasmic (HPA subcellular; HPA tissue IHC). The observations come from different applications. | Score the paraffin section using its visible cellular pattern and tissue controls. Use the separate IF/ICC guide for fluorescence interpretation; do not transfer an ICC-IF appearance into an IHC-P protocol expectation. |
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 look for staining in glandular cells (Medium; HPA: adrenal gland glandular cells). Use breast adipocytes as the negative tissue cell population (HPA: breast adipocytes, Not detected). The HPA adrenal row does not identify an internal negative cell population, so do not treat unstained neighboring cells as validated negatives (HPA: adrenal gland glandular cells).
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 a no-primary, secondary-only slide and a rabbit IgG isotype control matched to the primary antibody’s clonality, plus MAP2K1-knockout material as a biological negative (selected tissue-IHC caption: rabbit primary; standard IHC controls). Block endogenous peroxidase and check for background in the adrenal section when using HRP/DAB detection (selected tissue-IHC caption: HRP/DAB; standard chromogenic 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 tissue-IHC caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 for ovarian cancer tissue, but does not establish that retrieval is required for adrenal gland (selected tissue-IHC caption: EDTA pH 8.0). The supplied evidence does not establish whether frozen sections or IF are easier, or identify an adrenal-specific artefact (HPA: adrenal gland glandular cells; HPA subcellular: ICC-IF images).
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, compartment, cell identity and controls before comparing chromogenic signal across samples.
Which retrieval conditions should I start with for MAP2K1 paraffin IHC?
Start with heat-mediated retrieval in EDTA pH 8.0 (datasheet A00292). The documented paraffin-section example used this retrieval before incubating the catalog antibody at 2 μg/ml overnight at 4°C (datasheet A00292). If staining is weak, compare a modestly longer heating interval with the original condition on adjacent sections, while keeping antibody concentration and detection constant (standard IHC practice). Check that sections remain attached and morphology remains interpretable; tissue damage can make an apparent signal gain misleading (standard IHC practice). Record heating time and cooling conditions for each run so retrieval changes can be distinguished from staining variation (standard IHC practice).
Could fixation explain weak or uneven MAP2K1 staining?
The selected paraffin-section caption does not state a fixative, so MAP2K1 sensitivity to a particular fixative or fixation duration is unknown for this antibody (datasheet A00292). Document the fixative and processing history for each specimen, then compare sections with similar handling in the same staining run (standard IHC practice). If weak staining tracks a processing batch, examine morphology and a validated control section before changing antibody concentration (standard IHC practice). Keep the documented EDTA pH 8.0 retrieval as the starting condition while testing one processing variable at a time (datasheet A00292; standard IHC practice). Do not infer fixation tolerance from tissue expression patterns or MAP2K1 sequence features.
Where should MAP2K1 staining appear within cells?
Assess cytoplasmic staining first: tissue IHC shows a general cytoplasmic pattern, while subcellular imaging supports cytosol and plasma membrane localisation (HPA tissue IHC; HPA subcellular). MAP2K1 is also annotated in the nucleus and at cell-division structures, including centrosomes and the midbody (UniProt Q02750 localisation). A sharply membrane-associated signal can therefore merit review, but diffuse extracellular deposit does not match the annotated compartments (HPA subcellular; UniProt Q02750 localisation). Use a counterstain to distinguish cytoplasm from nuclei and tissue boundaries, and compare the distribution with an antibody-omission control (standard IHC practice). Score compartment and staining intensity separately rather than treating every brown pixel as equivalent (standard IHC practice).
Can isoforms or epitope accessibility change the IHC result?
MAP2K1 has 2 annotated isoforms, but the supplied antibody caption does not identify its recognised epitope or establish isoform selectivity (UniProt Q02750 isoforms; datasheet A00292). Its kinase domain spans residues 68–361, and reported modified residues include Ser218, Ser222, Thr286, Thr292 and Ser298 (UniProt Q02750 domains and modified residues). Those annotations alone cannot establish whether modification or splicing changes this antibody’s staining (UniProt Q02750; datasheet A00292). Check the antibody’s epitope documentation before making isoform-specific or phosphorylation-specific claims, and compare tissue staining with a separately validated assay if that distinction matters (standard IHC practice). Report the readout as MAP2K1 immunoreactivity when epitope specificity remains unresolved.
How should I investigate MAP2K1 localisation by multiplex IF?
Treat IF as a separately optimised application: the supplied antibody example documents chromogenic staining in paraffin sections, not an IF protocol (datasheet A00292). Pair MAP2K1 with a validated marker for the cell population under study; lung macrophages, for example, show medium tissue-IHC staining in the supplied atlas profile (HPA tissue IHC). Choose a fluorophore and imaging channel after checking the specimen’s autofluorescence, and include single-stain controls to assess bleed-through (standard IF practice). Because MAP2K1 lacks a transmembrane segment and has cytosolic as well as membrane-associated localisation, optimise permeabilisation for access to intracellular epitopes without assuming a membrane-facing epitope (UniProt Q02750 topology; HPA subcellular; standard IF practice).
How can I distinguish MAP2K1 signal from chromogenic background?
The documented example used 10% goat serum blocking, an anti-rabbit peroxidase secondary, and DAB development (datasheet A00292). Check an antibody-omission section for secondary-reagent deposits and use a peroxidase block where endogenous enzyme activity could contribute colour (standard IHC practice). Compare staining at tissue edges, folds and damaged regions with intact interior tissue; disproportionate colour in those areas warrants caution (standard IHC practice). If background persists, titrate the primary around the documented 2 μg/ml condition and keep development time identical across comparison sections (datasheet A00292; standard IHC practice). Judge any reduction against preservation of the expected cytoplasmic signal, rather than background alone (HPA tissue IHC).
What is a defensible way to quantify MAP2K1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and scoring compartment before measuring signal; the tissue-IHC profile describes general cytoplasmic expression (HPA tissue IHC). For comparable sections, report the percentage of positive cells and an H-score from intensity categories, with the scoring thresholds applied consistently across batches (standard IHC practice). Normalise positive counts to all evaluable cells of the same population, or report stained-cell density per mm² of viable tissue when cell counting is impractical (standard IHC practice). Exclude folds, necrosis and tissue edges using the same rules for every sample (standard IHC practice). Record retrieval and DAB development conditions because changes in either can shift measured intensity (standard IHC practice).
When should a positive MAP2K1 stain be questioned?
A credible result should have interpretable cellular boundaries and a distribution compatible with MAP2K1’s reported cytoplasmic pattern, with possible membrane-associated staining (HPA tissue IHC; HPA subcellular). Check cell identity: the supplied profile reports medium staining in lung macrophages but no detection in breast adipocytes, so those populations should not be pooled into one score (HPA tissue IHC). Uniform colour at cut edges, necrotic areas or sites of endogenous peroxidase activity can mimic positivity and needs control-section review (standard IHC practice). The tissue-IHC profile is marked approved while reporting low consistency with RNA expression, so a single stain should not establish abundance or pathway activity (HPA tissue IHC). Total MAP2K1 staining does not itself demonstrate phosphorylation at Ser218 or Ser222 (UniProt Q02750 modified residues; standard IHC interpretation).
Boster reagents
Best MAP2K1 / Dual specificity mitogen-activated protein kinase kinase 1 IHC Antibodies
The catalog includes IHC images of human ovarian cancer, kidney and lung, plus IF images; listed reactivity extends to Human, Mouse and Rat (catalog image captions and reactivity).
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 has IHC data from a human ovarian cancer paraffin section and IF data from TPC1 and U2OS cells; M00292-5 has IHC images of human kidney and lung (catalog image captions). M00292-1 has IHC data from a human kidney paraffin section and IF data from HeLa cells; M00292-2 has an IF figure whose caption does not identify the specimen (catalog image captions).
Which to pick: For tissue IHC, A00292 provides the most detailed starting conditions: its own paraffin-section caption specifies EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (A00292 IHC image caption). For IF/ICC, M00292-2 is listed for both applications and has an IF figure at 1:50, though its caption does not identify the specimen (M00292-2 catalog applications and IF image caption). For cross-species planning, monoclonal M00292-5 lists Human, Mouse and Rat reactivity, while its IHC images show human kidney and lung only; section preparation and fixative are unreported (M00292-5 catalog description, reactivity and 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 paraffin sections (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 2 isoforms; full-length chain, epitope coverage unknown (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). 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.