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MAP2K2 / Dual specificity mitogen-activated protein kinase kinase 2 · IHC design guide
Design Immunohistochemistry for MAP2K2
Plan chromogenic MAP2K2 IHC on paraffin sections using the IHC-validated antibody at 2–5 μg/ml (datasheet A00996-2). Appendix glandular cells show high staining, while heart cardiomyocytes are reported as undetected (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAP2K2 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 | General cytoplasmic staining, including glandular cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A00996-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Heart muscle+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00996-2) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation unannotated (UniProt) | |
| Isoform / epitope | No annotated isoforms or cleavage; no extracellular domain (UniProt) |
Section 1
Recommended MAP2K2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A00996-2) with the published HNSCC biopsy protocol (PMC10741104).
| Sample | Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A00996-2) |
| Fixation | Image fixative and duration unreported (datasheet A00996-2); 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 A00996-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A00996-2) |
| Primary antibody | Rabbit anti-MAP2K2, 2-5μg/ml (datasheet A00996-2) |
| Primary incubation | Overnight at 4 °C (datasheet A00996-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A00996-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MAP2K2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet A00996-2); the published citrate retrieval is an alternative to evaluate (PMC10741104).
Section 2
What Is the Expected MAP2K2 Staining Pattern?
MAP2K2 should appear mainly cytoplasmic in paraffin-section IHC, including strong staining in appendix and colon glandular cells and bone marrow hematopoietic cells (HPA: general cytoplasmic expression; High in these cells). UniProt also lists membrane localization, possibly regulated by KSR1 interaction, and reports no transmembrane segment (UniProt P36507: subcellular location and topology). Treat intensity expectations as guides: HPA rates tissue IHC reliability Supported, with medium consistency between staining and RNA data (HPA: reliability).
| Cytoplasmic staining is strong in appendix or colon glandular cells, with recognizable cell boundaries. | This matches the reported compartment and high-staining cell populations (HPA: general cytoplasmic expression; High in appendix and colon glandular cells). Compare signal within the intended cell population, because adjacent cells need not share its staining level (general IHC interpretation). |
| Signal appears predominantly nuclear, or as sharply outlined plasma membranes without convincing cytoplasm. | Recheck specificity and imaging or scoring: HPA describes general cytoplasmic tissue staining, while UniProt lists cytoplasm and potentially regulated membrane localization (HPA: tissue profile; UniProt P36507: subcellular location). A membrane-associated component alone is therefore insufficient evidence of an artefact. |
| Cardiomyocytes stain strongly in heart muscle despite an otherwise plausible slide. | That conflicts with the reported absence of detectable cardiomyocyte staining (HPA: heart muscle, cardiomyocytes Not detected). Consider cross-reactivity or chromogenic detection activity, then compare a control lacking primary antibody (general IHC practice). Do not score other heart cell types from this cardiomyocyte finding. |
| Brown deposit covers tissue broadly, including spaces or structures without clear cellular outlines. | A diffuse deposit is difficult to assign to MAP2K2-positive cells (general IHC interpretation). Review blocking, washing and chromogen development, and use a control lacking primary antibody to assess detection background (general IHC practice). |
| Appendix glandular cells or bone marrow hematopoietic cells show no convincing cytoplasmic signal. | These are reported high-staining populations (HPA: High in appendix glandular cells and bone marrow hematopoietic cells). A negative run is inconclusive until section quality, retrieval and detection controls are checked (general IHC practice); HPA does not establish MAP2K2-specific fixation sensitivity. |
💡Expected MAP2K2 appearanceCall a section positive when defined glandular or hematopoietic cells show clear cytoplasmic chromogen above local background, with strong signal plausible in appendix, colon or bone marrow (HPA: general cytoplasmic expression; High in these cells); widespread acellular deposit or dominant nuclear-only staining warrants investigation (general IHC interpretation; HPA: tissue profile).
| Cell population and tissue | HPA reports High staining in appendix, colon and duodenal glandular cells, bone marrow hematopoietic cells and cerebellar Purkinje cells; cardiomyocytes and skeletal and smooth muscle cells are Not detected (HPA: tissue IHC). Evaluate the named cells, rather than assigning one score to the whole section (general IHC interpretation). |
| Compartment and topology | The tissue profile is generally cytoplasmic; UniProt lists cytoplasm and membrane, with membrane localization possibly regulated by KSR1 interaction (HPA: tissue profile; UniProt P36507: subcellular location). No transmembrane segment is annotated, so an exclusively crisp surface pattern needs further assessment (UniProt P36507: topology; general IHC interpretation). |
| Evidence strength and antibody comparison | HPA calls tissue IHC Supported and describes medium consistency with RNA data; both listed HPA antibodies have Supported IHC status (HPA: reliability; HPA051993 and CAB003835 IHC validation). Those ratings describe the HPA observations and do not independently validate a different catalog antibody (general assay interpretation). |
| Antigen form and epitope limits | UniProt annotates one 1–400 chain, no signal peptide or propeptide, and no listed isoforms (UniProt P36507: processing and isoforms). Phosphorylated residues are annotated, but the supplied evidence gives no IHC antibody epitope or MAP2K2-specific retrieval response; do not infer a required retrieval condition from those annotations (UniProt P36507: modified residues; supplied-source limit). |
| IF/ICC Q: what localization should appear? | A: HPA reports mainly cytosolic IF signal, with additional sperm mid-piece, principal-piece and end-piece localization (HPA: ICC-IF subcellular summary). This cross-check informs compartment interpretation; the present guide's primary readout is chromogenic paraffin-section IHC (application scope). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known high-staining cells are blank. | The run may have inadequate retrieval or failed antibody or detection steps (general IHC practice); target-specific fixation sensitivity is unreported in the supplied evidence. | Check section integrity, the IHC-validated antibody's IHC-P instructions, retrieval and detection controls; rerun a reported high-staining population such as appendix glandular cells (HPA: High in appendix glandular cells; general IHC practice). |
| Chromogen is widespread and cell outlines are hard to distinguish. | Excess detection background or endogenous activity can obscure cellular staining in chromogenic IHC (general IHC practice). | Review blocking, washes and chromogen development; compare a control lacking primary antibody, especially before interpreting marrow signal (general IHC practice; HPA: High in bone marrow hematopoietic cells). |
| Cardiomyocytes appear strongly positive. | Cross-reactivity or detection background is possible because cardiomyocytes are reported Not detected (HPA: heart muscle IHC; general IHC interpretation). | Confirm the stained cell identity, inspect a control lacking primary antibody and compare a reported positive tissue in the same run (general IHC practice; HPA: appendix glandular cells High). |
| Signal is predominantly nuclear. | The compartment disagrees with HPA's general cytoplasmic tissue profile and UniProt's cytoplasm and membrane annotation (HPA: tissue profile; UniProt P36507: subcellular location). | Review counterstain and focal plane, check detection background, then reassess using a reported positive cell population (general IHC practice; HPA: appendix glandular cells High). |
| Only a thin cell-edge signal is visible. | UniProt permits regulated membrane association, but the expected tissue IHC profile is generally cytoplasmic (UniProt P36507: subcellular location; HPA: tissue profile). | Check whether cytoplasm is also stained, compare local background and confirm the pattern with appropriate controls before scoring it as MAP2K2 (general IHC interpretation). |
| A low- or absent-staining population is used as the sole run control. | HPA reports Low staining in hippocampal neuronal cells and Not detected staining in skeletal myocytes, so either gives limited evidence that a negative assay worked (HPA: tissue IHC). | Include a reported high-staining cell population as a positive comparison and score each named population separately (HPA: appendix glandular cells High; general IHC practice). |
Experiment evidence
IHC & IF images showing MAP2K2 staining
A00996-2 · IHCIHC analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (A00996-2). MEK2/MAP2K2 was detected in paraffin-embedded section of human recta
A00996-2 · IHCIHC analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (A00996-2). MEK2/MAP2K2 was detected in paraffin-embedded section of rat brain t
A00996-2 · IFIF analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (A00996-2). MEK2/MAP2K2 was detected in immunocytochemical section of U20S cells.
M00996 · IHCIHC analysis of MAP2K2 using anti-MAP2K2 antibody (M00996). MAP2K2 was detected in a paraffin-embedded section of human stomach cancer tissu
M00996-3 · IHCIHC analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (M00996-3). MEK2/MAP2K2 was detected in paraffin-embedded section of human renal
M00996-3 · IFIF analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (M00996-3). MEK2/MAP2K2 was detected in immunocytochemical section of MCF-7 cells
M00996-2 · IFIF analysis of MEK2/MAP2K2 using anti-MEK2/MAP2K2 antibody (M00996-2). MEK2/MAP2K2 was detected in immunocytochemical section of MCF-7 cells
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000126934-MAP2K2/tissue/cerebral+cortex#imid_1315748
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000126934-MAP2K2/tissue/colon#imid_1315674
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000126934-MAP2K2/tissue/liver#imid_1315667
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000126934-MAP2K2/tissue/kidney#imid_1315676
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB003835; green = MAP2K2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB003835; green = MAP2K2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAP2K2 IHC & IF
🧪Run colon first: its glandular cells should show strong staining (HPA: High in colon glandular cells). Use skeletal muscle as the negative tissue; its myocytes should lack detectable staining (HPA: Not detected in skeletal-muscle myocytes). On the colon slide, any unstained non-glandular cells should show counterstain without brown DAB deposit; use them as a background reference, not as a validated biological negative (HPA: High in colon glandular cells).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Heart muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAP2K2 in A-431, U-251MG, U2OS, MCF-7, Sperm, NIH 3T3, with annotated localisation: Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit IgG isotype control for the rabbit primary (selected-SKU caption: rabbit anti-MAP2K2 antibody). Confirm target dependence with MAP2K2 knockout tissue or cells processed alongside the test specimen; for the biotin-based DAB workflow, quench endogenous peroxidase and check for endogenous biotin signal in colon (selected-SKU caption: biotinylated secondary, streptavidin-biotin complex and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state a fixative (selected-SKU caption: fixative not stated). The documented IHC workflow uses heat-mediated retrieval in EDTA at pH 8.0, but whether MAP2K2 staining requires that retrieval condition is unreported (selected-SKU caption: EDTA heat retrieval). Frozen sections have no supplied comparison, and IF cannot be judged easier from the supplied evidence; ICC-IF images support mainly cytosolic localisation (HPA: Cytosol, enhanced).
HPA tissue IHC evidence for MAP2K2
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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced MAP2K2 IHC Tips
Troubleshoot MAP2K2 staining in paraffin sections by checking retrieval, cytoplasmic localisation, controls, and how staining is scored.
How should I optimise retrieval when MAP2K2 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A00996-2). The documented paraffin-section example then used 2 µg/mL primary antibody overnight at 4°C, so keep those conditions steady while assessing retrieval (datasheet A00996-2). Compare a short and a longer heating period on matched sections, recording tissue preservation and cytoplasmic signal in glandular cells (standard IHC practice; HPA: high in colon glandular cells). If staining remains weak, test another retrieval buffer or pH as a fallback, with a matched control section (standard IHC practice). Excess heating can damage morphology, making apparent gains in signal difficult to interpret (standard IHC practice).
Could fixation explain inconsistent MAP2K2 staining between paraffin blocks?
The selected paraffin-section caption does not state a fixative, so MAP2K2-specific sensitivity to fixation is unknown (datasheet A00996-2). Record each block’s fixative and fixation duration, then compare matched sections with identical EDTA pH 8.0 retrieval and staining conditions (datasheet A00996-2; standard IHC practice). Include a previously stained control block in each run to separate block variation from changes in reagents or processing (standard IHC practice). If morphology and several other antigens also stain poorly, investigate tissue processing before assigning the weak MAP2K2 result to fixation (standard IHC practice). Do not infer a fixation effect from cytoplasmic localisation or tissue expression patterns (UniProt P36507; HPA: tissue IHC).
Is membrane-associated MAP2K2 staining credible in chromogenic IHC?
Expect predominantly cytoplasmic staining: tissue IHC reports general cytoplasmic expression, and cell imaging places MAP2K2 mainly in the cytosol (HPA: tissue IHC; HPA: subcellular). Membrane-associated signal can be plausible because UniProt also lists membrane localisation, probably regulated through KSR1 interaction (UniProt P36507: subcellular location). MAP2K2 has no transmembrane segment, so an isolated, sharply membranous pattern merits closer scrutiny (UniProt P36507: topology). Compare the pattern with cytoplasmic staining in an HPA-supported cell population and with a no-primary control processed alongside it (HPA: high in colon glandular cells; standard IHC practice). Record mixed cytoplasmic and peripheral staining separately during review (standard IHC practice).
What epitope issues matter when comparing MAP2K2 IHC antibodies?
The supplied record lists 0 isoforms and a single 1–400 chain, so isoform switching is not an evidence-based explanation for discordant staining here (UniProt P36507: isoforms and processing). Its kinase domain spans residues 72–369, and the record lists modified residues including RAF-associated phosphoserine alternatives at 222 and 226 (UniProt P36507: domains and modified residues). Check each antibody’s disclosed immunogen and phospho-specificity before comparing results; neither is established for the selected antibody in the supplied caption (datasheet A00996-2; standard IHC practice). An antibody recognising total MAP2K2 cannot, without validation, establish its phosphorylation state (standard IHC practice). Keep retrieval and detection conditions matched when comparing antibodies (standard IHC practice).
How can I check an IHC pattern with MAP2K2 immunofluorescence?
Use IF/ICC as a separate optimisation, since the supplied antibody example documents paraffin-section chromogenic IHC rather than an IF protocol (datasheet A00996-2). Multiplex MAP2K2 with a marker identifying the expected cell population, such as glandular cells in colon, and compare the resulting cytosolic pattern (HPA: high in colon glandular cells; HPA: mainly cytosolic). Choose spectrally separated fluorophores and place the weaker signal in a channel with low measured tissue autofluorescence (standard IF practice). MAP2K2 has no transmembrane segment and is mainly cytosolic, so permeabilise cells to expose intracellular epitopes, then optimise detergent strength against morphology (UniProt P36507: topology; HPA: subcellular; standard IF practice). Include single-label and no-primary controls for bleed-through and background (standard IF practice).
How do I reduce diffuse brown background without losing MAP2K2 signal?
The documented paraffin-section procedure blocked with 10% goat serum, used 2 µg/mL rabbit primary overnight at 4°C, and developed with biotin-based detection and DAB (datasheet A00996-2). Start by checking a no-primary section and shortening DAB development if diffuse colour appears there (standard IHC practice). Apply an endogenous peroxidase block before DAB detection, and assess whether the biotin-based system contributes background with an appropriate detection control (standard IHC practice; datasheet A00996-2). Titrate primary concentration around the documented setting while holding retrieval and imaging conditions steady (datasheet A00996-2; standard IHC practice). Preserve interpretable cytoplasmic signal in positive cell populations when selecting the final condition (HPA: general cytoplasmic expression; standard IHC practice).
What should I score when comparing MAP2K2 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score cytoplasmic staining within a predefined cell population, since the tissue profile describes general cytoplasmic expression (HPA: tissue IHC). An H-score can combine the percentage of cells at each intensity level with intensity grades, while percentage positive alone is useful when intensity is unreliable (standard IHC practice). Define positivity against a no-primary control, apply one scoring threshold across batches, and report the number of evaluable cells or tissue area (standard IHC practice). Normalise positive-cell counts to all evaluable cells of the same type, or report positive-cell density per mm² of viable tissue (standard IHC practice). Exclude necrotic and damaged areas consistently before comparing groups (standard IHC practice).
How can I distinguish genuine MAP2K2 staining from artefact?
A credible result shows predominantly cytoplasmic signal in intact cells, consistent with tissue IHC and cell imaging (HPA: general cytoplasmic expression; HPA: mainly cytosolic). High staining in colon glandular cells offers a documented comparison, while cardiomyocytes were reported as not detected; neither pattern alone validates an individual section (HPA: tissue IHC). Be cautious with isolated nuclear or sharply membranous colour, although regulated membrane association is possible through KSR1 (UniProt P36507: subcellular location and topology). Check whether staining follows cut edges, necrosis, or damaged tissue, and compare with a no-primary section for nonspecific or endogenous enzyme colour (standard IHC practice). Interpret intensity in light of HPA’s Supported reliability rating and medium RNA–staining consistency (HPA: tissue IHC).
Boster reagents
Best MAP2K2 / Dual specificity mitogen-activated protein kinase kinase 2 IHC Antibodies
Catalog antibodies have IHC images from human paraffin sections and mouse and rat brain sections, plus IF images from human cells (catalog image captions: A00996-2, M00996, M00996-2, M00996-3).
Real IHC data
Anti-MEK2/MAP2K2 Antibody ®
Cat # A00996-2
Real IHC data
Anti-MEK2 MAP2K2 Rabbit Monoclonal Antibody
Cat # M00996
Real IHC data
Anti-MEK2/MAP2K2 Antibody ® (monoclonal, 2H4)
Cat # M00996-3
Real IF data
Anti-MEK2/MAP2K2 Antibody ® (monoclonal, 2B4)
Cat # M00996-2
A00996-2 has IHC images from human rectal cancer and mouse and rat brain paraffin sections; M00996 has an IHC image from a human stomach cancer paraffin section (catalog IHC image captions: A00996-2, M00996). M00996-3 has an IHC image from a human renal clear cell carcinoma paraffin section and an IF image from MCF-7 cells; M00996-2 has an IF image from MCF-7 cells and does not list IHC as an application (catalog applications and image captions: M00996-3, M00996-2).
Which to pick: For human tissue IHC, choose A00996-2, M00996, or M00996-3 according to the relevant paraffin section image; each caption reports EDTA retrieval at pH 8.0, while the fixative is unreported (catalog IHC image captions: A00996-2, M00996, M00996-3). For IF/ICC, M00996-2 has a human MCF-7 cell IF image and is a mouse monoclonal antibody; M00996-3 also has a human MCF-7 cell IF image and lists IHC (catalog applications, clone and IF image captions: M00996-2, M00996-3). For cross-species tissue IHC, A00996-2 has paraffin section images from human, mouse, and rat samples; M00996-3 lists reactivity with all three species, but its IHC image shows only human tissue (catalog reactivity and IHC image captions: A00996-2, M00996-3).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P36507 (MP2K2_HUMAN, Dual specificity mitogen-activated protein kinase kinase 2).
- Human Protein Atlas. MAP2K2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MAP2K2 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the mid piece, principal piece and end piece..
- Human Protein Atlas. MAP2K2 antibody validation summary (2 antibodies).
- The Prognosis Performance of a Neutrophil- and Lymphocyte-Associated Gene Mutation Score in a Head and Neck Cancer Cohort. Biomedicines 2023 — PMC10741104.
- Development and validation of cuproptosis-related lncRNAs associated with pancreatic cancer immune microenvironment based on single-cell. Frontiers in immunology 2023 — PMC10563513.
- Expression-Based Diagnosis, Treatment Selection, and Drug Development for Breast Cancer. International journal of molecular sciences 2023 — PMC10342177.
- Post-transcriptional modification of m(6)A methylase METTL3 regulates ERK-induced androgen-deprived treatment resistance prostate cancer. Cell death & disease 2023 — PMC10126012.
- PubMed PMID:8388392 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:17116858 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAP2K2 (Dual specificity mitogen-activated protein kinase kinase 2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A00996-2). Validated IHC antibody: A00996-2 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Heart muscle. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A00996-2); 2. Fixation: Image fixative and duration unreported (datasheet A00996-2); 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 A00996-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A00996-2); 8. Primary antibody: Rabbit anti-MAP2K2, 2-5μg/ml (datasheet A00996-2); 9. Primary incubation: Overnight at 4 °C (datasheet A00996-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A00996-2); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No annotated isoforms or cleavage; no extracellular domain (UniProt); Regulation: Expression regulation unannotated (UniProt). Evidence: UniProt P36507, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the mid…, Datasheet A00996-2, PMC10741104. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAP2K2/immunohistochemistry-ihc-guide.