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MAPK14 / Mitogen-activated protein kinase 14 · IHC design guide
Design Immunohistochemistry for MAPK14
Plan MAPK14 staining in paraffin sections using the cytoplasmic and nuclear tissue pattern reported by HPA (HPA tissue IHC). The guide covers antibody selection, staining controls and interpretation of total versus phospho-specific signal (UniProt; datasheet: A00176-2, A00176T180Y182).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAPK14 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 and nuclear staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and nuclear staining in multiple cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet M00176-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 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 M00176-1) | |
| Caveat | Total MAPK14 staining does not establish activation (UniProt) | |
| Regulation | Cytokines and stress activate MAPK14 (UniProt) | |
| Isoform / epitope | 5 isoforms; check epitope coverage for the chosen antibody (UniProt) |
Section 1
Recommended MAPK14 IHC & IF Protocols
Start with the catalog antibody’s citrate pH 6 IHC-P protocol (datasheet: M00176-1). The published methods below cover colorectal, liver, patient and mouse, and mouse carotid samples (PMC8818961; PMC7967205; PMC7519103; PMC6377391).
| Sample | Paraffin-embedded tissue sections; fixative not specified (datasheet M00176-1; sample unspecified) |
| Fixation | Image fixative and duration unreported (datasheet M00176-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet M00176-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00176-1) |
| Primary antibody | Rabbit monoclonal (clone EGC-13) anti-MAPK14, 1:50 recommended; image 1ug/ml (datasheet M00176-1) |
| Primary incubation | Overnight at 4 °C (datasheet M00176-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M00176-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MAPK14-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic and nuclear expression. No signal in the no-primary control. |
💡Decision noteTry heat-mediated citrate pH 6 retrieval first (datasheet: M00176-1). The published citrate conditions range from boiling for 10 min to incubation for 1 h (PMC8818961; PMC7519103).
Section 2
What Is the Expected MAPK14 Staining Pattern?
MAPK14 is expected in cytoplasm and nuclei, with tissue dependent staining in defined cell populations (UniProt Q16539 subcellular location; HPA tissue IHC: general cytoplasmic and nuclear expression). Its lack of a transmembrane segment makes a membrane restricted pattern unexpected (UniProt Q16539 topology). HPA rates the tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression; interpret individual cells and controls accordingly (HPA tissue IHC: reliability).
| Cytoplasmic and nuclear signal in cerebral cortical neurons or bone marrow hematopoietic cells. | This fits the reported compartments and medium staining in those cell populations (UniProt Q16539 subcellular location; HPA tissue IHC: cerebral cortex and bone marrow). Compare cells within the section; the HPA rating describes observed staining, not a guaranteed intensity for every specimen (HPA tissue IHC: Approved, medium consistency). |
| Signal appears exclusively along cell borders, with little cytoplasmic or nuclear staining. | A membrane restricted pattern conflicts with the reported intracellular locations and absence of a transmembrane segment (UniProt Q16539 subcellular location and topology). Check the antibody and detection controls before assigning it to MAPK14; a cell border signal alone does not establish target localization (general IHC practice). |
| Strong signal appears in adipocytes or smooth muscle cells. | HPA reports MAPK14 as not detected in those cell populations (HPA tissue IHC: adipose tissue and smooth muscle). Investigate antibody cross reactivity or endogenous detection activity with appropriate controls (general IHC practice). A discordant result needs validation because HPA's Approved profile has only medium staining–RNA consistency (HPA tissue IHC: reliability). |
| Color spreads across tissue or appears similarly in cells and surrounding spaces. | That distribution is difficult to assign to the reported cytoplasmic and nuclear compartments (UniProt Q16539 subcellular location; HPA tissue IHC: profile). Review the no primary control, blocking, washes and chromogen development to distinguish background from cell associated staining (general IHC practice). Do not score a diffuse deposit as a positive cell (general IHC practice). |
| No signal is seen in cerebral cortical neurons on a test section. | HPA reports medium neuronal staining in cerebral cortex, so absence warrants a technical check (HPA tissue IHC: cerebral cortex). Confirm that the expected cells are present, then review the antibody's IHC-P instructions and detection controls (general IHC practice). An HPA positive example is a reference pattern, not proof that every specimen must stain (HPA tissue IHC: Approved, medium consistency). |
💡Expected MAPK14 appearanceCall a result positive when intracellular cytoplasmic and/or nuclear signal is discernible in an HPA listed cell population, such as medium staining in cerebral cortical neurons; isolated cell border staining or uniform tissue haze is suspect (HPA tissue IHC: profile and cerebral cortex; UniProt Q16539 topology; general IHC practice).
| Compartment and topology | MAPK14 is reported in cytoplasm and nucleus and has no transmembrane segment (UniProt Q16539 subcellular location and topology). Assess staining inside identifiable cells; a membrane restricted result requires scrutiny rather than automatic acceptance as the expected pattern (general IHC practice). |
| Tissue and cell context | HPA lists medium staining in cortical neurons, marrow hematopoietic cells and several epithelial or glandular populations, but low staining in lung alveolar cells and no detection in adipocytes (HPA tissue IHC: positive, low and negative lists). Select comparison regions by cell type, not tissue name alone (general IHC practice). |
| Evidence strength and antibody choice | The tissue profile is Approved with medium staining–RNA consistency; CAB010285 and CAB040578 are IHC Approved, while HPA051825 is ICC Supported with no IHC status supplied (HPA tissue IHC: reliability; HPA antibodies). These labels do not establish that every antibody will reproduce every listed tissue pattern (HPA antibody validation summary). |
| Isoforms and modifications | UniProt lists 5 isoforms and multiple modified residues, including phosphorylation and acetylation (UniProt Q16539 isoforms and modified residues). The payload gives no epitope for the IHC antibodies, so neither isoform coverage nor modification dependent staining can be inferred from these entries (HPA antibodies; UniProt Q16539). |
| IF localization, separate application | HPA ICC-IF places MAPK14 mainly in supported nuclear speckles, with supported cytosolic localization (HPA subcellular ICC-IF). This higher resolution IF observation can inform interpretation, but an IHC paraffin section need not resolve speckles (general microscopy practice); the supplied IHC evidence describes general nuclear and cytoplasmic staining (HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| No staining in an HPA positive cell population. | The expected cells may be absent, or the IHC workflow or detection may have failed (general IHC practice); HPA reports medium staining in cortical neurons and marrow hematopoietic cells (HPA tissue IHC: positive list). | Verify the cell population, run a known positive section, and review the IHC-validated antibody's instructions plus detection controls (general IHC practice). Avoid declaring a specimen negative from a failed control (general IHC practice). |
| Diffuse color obscures cell boundaries. | Background from blocking, washing or chromogen development can obscure intracellular signal (general IHC practice). | Compare a no primary control and adjust the general IHC blocking, washing or development workflow as indicated (general IHC practice). Score only identifiable cellular staining in the reported compartments (HPA tissue IHC: profile; general IHC practice). |
| Adipocytes or smooth muscle cells stain prominently. | These populations are listed as not detected, so cross reactivity or endogenous detection activity is plausible (HPA tissue IHC: adipose tissue and smooth muscle; general IHC practice). | Check no primary and detection controls, then verify the antibody's IHC validation before interpreting the signal as MAPK14 (general IHC practice; HPA antibodies: IHC status). |
| Only cell borders stain. | The pattern conflicts with MAPK14's reported cytoplasmic and nuclear location and lack of a transmembrane segment (UniProt Q16539 subcellular location and topology). | Inspect control sections and detection artifacts, and seek reproducible intracellular staining in an HPA listed cell population before calling the result positive (general IHC practice; HPA tissue IHC: positive list). |
| Staining varies between cell types within one section. | HPA reports different levels by cell population, including medium cortical neuronal staining, low lung alveolar staining and no detection in adipocytes (HPA tissue IHC: positive, low and negative lists). | Record compartment and intensity separately for each identifiable cell type, using the corresponding HPA cell population as the comparison (HPA tissue IHC: tissue entries; general IHC practice). |
| IF/ICC Q: Must a paraffin IHC section show nuclear speckles? | HPA's supported nuclear speckle observation comes from ICC-IF, whereas its tissue IHC profile reports general nuclear and cytoplasmic expression (HPA subcellular ICC-IF; HPA tissue IHC: profile). | A: Judge IHC by recognizable intracellular staining and its cell type; consult the separate IF/ICC guide for that application (HPA tissue IHC: profile; general microscopy practice). |
Experiment evidence
IHC & IF images showing MAPK14 staining
M00176-1 · IHCIHC analysis of p38 MAPK using anti-p38 MAPK antibody (M00176-1) on human brain. p38 MAPK was detected in paraffin-embedded section. Heat me
M00176-1 · IHCIHC analysis of p38 MAPK using anti-p38 MAPK antibody (M00176-1) on human brain. p38 MAPK was detected in paraffin-embedded section. Heat me
M00176-1 · IFIF analysis of immunocytochemical section of Hela cells using anti-p38 MAPK antibody (M00176-1) p38 MAPK was detected in immunocytochemical
A00176T180Y182 · IHCImmunohistochemical analysis of paraffin-embedded Human-lung tissue. 1, p38 (phospho Thr180/Y182) Polyclonal Antibody was diluted at 1:200 (
A00176T180Y182 · IHCImmunohistochemical analysis of paraffin-embedded Human-lung-cancer tissue. 1, p38 (phospho Thr180/Y182) Polyclonal Antibody was diluted at
A00176T180Y182 · IFImmunofluorescence analysis of Hela cell. 1, p38 (phospho Thr180/Y182) Polyclonal Antibody (green) was diluted at 1:200 (4° overnight). 2, G
A00176-2 · IHCIHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2). p38 alpha/MAPK14 was detected in a paraffin-embedded secti
A00176-2 · IHCIHC analysis of p38 alpha/MAPK14 using anti-p38 alpha/MAPK14 antibody (A00176-2). p38 alpha/MAPK14 was detected in a paraffin-embedded secti
M00176 · IFImmunofluorescent analysis of Hela cells, using p38 MAPK Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000112062-MAPK14/tissue/cerebral+cortex#imid_18439903
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000112062-MAPK14/tissue/colon#imid_18439829
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000112062-MAPK14/tissue/liver#imid_18439822
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000112062-MAPK14/tissue/kidney#imid_18439831
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA051825; green = MAPK14, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA051825; green = MAPK14, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAPK14 IHC & IF
🧪Run cerebral cortex first and confirm staining in neuronal cells (HPA: Medium in cerebral cortex neuronal cells); the selected catalog antibody also has a paraffin-section image from human brain (catalog caption: M00176-1). Run adipose tissue as the negative tissue and assess adipocytes (HPA: Not detected in adipocytes); on the positive slide, neighboring unstained cells and clear background provide internal negative areas, without assuming a particular cell type must be negative (standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAPK14 in A-431, U-251MG, U2OS, ASC52telo, NIH 3T3, with annotated localisation: Nuclear speckles (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype-matched rabbit IgG control matched to the primary antibody’s clonality (catalog caption: rabbit primary; standard IHC practice); use MAPK14-knockout material or an immunizing-peptide block, if available, as a biological specificity control (standard IHC practice). For the brain slide, block endogenous peroxidase and check endogenous biotin background before interpreting the biotin–streptavidin/DAB signal (catalog caption: biotinylated secondary and SABC/DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected tissue-IHC caption does not state the fixative (catalog caption: M00176-1). The demonstrated paraffin-section workflow uses heat retrieval in citrate buffer at pH 6 for 20 minutes; retrieval dependence beyond that example is unreported (catalog caption: M00176-1). No supplied matched comparison establishes that frozen sections or IF are easier; for brain IF, check lipofuscin autofluorescence with an unstained section (standard IF practice).
HPA tissue IHC evidence for MAPK14
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | Medium | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAPK14 IHC Tips
Troubleshoot MAPK14 staining in paraffin sections by checking retrieval, controls, compartment assignment and cell-level scoring before interpreting signal as pathway activity.
How should I adjust retrieval when MAPK14 staining is weak or uneven?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes in paraffin sections (datasheet M00176-1). Compare a known positive section and an adjacent no-primary control in the same run, keeping section thickness, heating and detection constant (standard IHC practice). The selected image used 1 µg/mL primary antibody overnight at 4°C, so match those conditions before changing retrieval (datasheet M00176-1). If staining remains weak, test a different retrieval condition on serial sections while checking whether tissue morphology or background deteriorates (standard IHC practice). Record cytoplasmic and nuclear signal separately because both compartments are plausible for MAPK14 (UniProt Q16539 subcellular location).
Could fixation explain weak MAPK14 staining in paraffin sections?
The selected paraffin-section caption does not state a fixative, so target-specific fixation sensitivity for MAPK14 is unknown (datasheet M00176-1). Document the specimen’s actual fixative and fixation interval, then compare sections processed under matched retrieval and detection conditions (standard IHC practice). Begin with citrate pH 6 retrieval for 20 minutes when using the catalog antibody’s documented method (datasheet M00176-1). If signal differs across processing batches, inspect morphology and compare a shared control section before attributing the difference to MAPK14 abundance (standard IHC practice). Do not infer a fixation effect from MAPK14’s tissue distribution or intracellular location (HPA tissue IHC; UniProt Q16539 subcellular location).
Is nuclear staining compatible with MAPK14, and how should I check it?
Score cytoplasmic and nuclear staining separately because MAPK14 is reported in both compartments (UniProt Q16539 subcellular location; HPA tissue IHC). Use a counterstain and inspect intact cells at high magnification to distinguish nuclear signal from overlying cytoplasm or precipitate (standard IHC practice). HPA subcellular imaging supports nuclear speckles and cytosol, but that fine pattern need not resolve in chromogenic tissue sections (HPA subcellular; standard IHC practice). Compare serial sections with a no-primary control and apply the same pH 6 retrieval across them (datasheet M00176-1; standard IHC practice). A compartment shift alone does not establish activation; the catalog image documents p38 MAPK detection, not phosphorylation-specific detection (datasheet M00176-1).
Can this stain distinguish MAPK14 isoforms or a phosphorylated epitope?
MAPK14 has 5 listed isoforms, including CSBP2, CSBP1, Mxi2 and Exip (UniProt Q16539 isoforms). The supplied caption identifies a p38 MAPK antibody but gives no immunogen, mapped epitope or isoform cross-reactivity (datasheet M00176-1). Therefore, treat staining as antibody-dependent MAPK14-associated signal until epitope specificity is established with appropriate controls (datasheet M00176-1; standard IHC practice). MAPK14 carries reported modified residues, but total-antibody staining cannot assign any particular phosphorylation state without phospho-specific validation (UniProt Q16539 modified residues; standard IHC practice). Compare candidate isoform or phospho reagents on matched serial sections, and document each reagent’s independent validation before interpreting differences (standard IHC practice).
How should I adapt MAPK14 assessment to multiplex tissue IF?
Treat IF as a separately validated application: the selected antibody evidence describes chromogenic staining of paraffin sections, not IF performance (datasheet M00176-1). Multiplex MAPK14 with a validated marker identifying the cell population being scored, and check each channel alone for bleed-through (standard IF practice). Choose a fluorophore whose emission avoids the specimen’s strongest autofluorescence, using an unstained section to assess that background (standard IF practice). Because MAPK14 is cytoplasmic and nuclear with no transmembrane segment, permeabilisation should permit access to intracellular epitopes if the antibody requires it (UniProt Q16539 subcellular location and topology; standard IF practice). Compare permeabilised and unpermeabilised controls, then score nuclear and cytoplasmic signal separately rather than assuming IF matches DAB localisation (standard IF practice).
What causes diffuse brown background in MAPK14 IHC?
First inspect a no-primary section for endogenous enzyme activity, nonspecific secondary binding and chromogen deposits (standard IHC practice). The documented workflow used 10% goat serum, a biotinylated secondary, a streptavidin–biotin complex and DAB (datasheet M00176-1). Include an appropriate peroxidase block and consider whether endogenous biotin contributes to signal with that detection chemistry (standard IHC practice; datasheet M00176-1). If background persists, compare shorter chromogen development and more thorough washes while holding retrieval at citrate pH 6 constant (standard IHC practice; datasheet M00176-1). Assess background beside intact positive cells, since genuine MAPK14 staining may occur in cytoplasm and nucleus (HPA tissue IHC; UniProt Q16539 subcellular location).
How should I quantify MAPK14 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record % positive cells and intensity or calculate an H-score using consistent thresholds (standard IHC practice). Report nuclear and cytoplasmic scores separately because both are documented MAPK14 locations (HPA tissue IHC; UniProt Q16539 subcellular location). For spatial comparisons, count positive cells per mm² of viable, annotated tissue rather than dividing by the entire slide area (standard IHC practice). Normalise to the number of evaluable cells or viable tissue area within the same cell population, and keep exposure-independent DAB development consistent (standard IHC practice). Use the same control section and retrieval conditions across batches before comparing scores (standard IHC practice; datasheet M00176-1).
How can I distinguish true MAPK14 staining from section artefacts?
Prioritise staining in intact cells with plausible cytoplasmic or nuclear distribution, as reported for MAPK14 (UniProt Q16539 subcellular location; HPA tissue IHC). Compare the candidate signal with a no-primary control and examine edge effects, necrotic regions, precipitate and endogenous peroxidase activity (standard IHC practice). HPA reports medium staining in cerebral-cortex neuronal cells but no detection in adipocytes, providing examples for cell-aware comparison rather than universal controls (HPA tissue IHC). Recheck unexpected membrane-only staining because MAPK14 has no transmembrane segment, while recognising that morphology alone cannot establish specificity (UniProt Q16539 topology; standard IHC practice). Finally, do not equate total MAPK14 DAB intensity with kinase activation or a specific isoform without separately validated assays (datasheet M00176-1; UniProt Q16539 isoforms; standard IHC practice).
Boster reagents
Best MAPK14 / Mitogen-activated protein kinase 14 IHC Antibodies
IHC images show human brain, lung, and breast cancer paraffin sections and mouse and rat brain paraffin sections (catalog IHC captions); IF images show HeLa cells (catalog IF captions).
Real IHC data
Anti-p38 MAPK MAPK14 Rabbit Monoclonal Antibody
Cat # M00176-1
Real IHC data
Anti-Phospho-p38 (T180/Y182) MAPK14 Antibody
Cat # A00176T180Y182
Real IHC data
Anti-p38 alpha/MAPK14 Antibody ®
Cat # A00176-2
Real IF data
Anti-p38 MAPK14 Rabbit Monoclonal Antibody
Cat # M00176
M00176-1 has human brain paraffin IHC and HeLa IF images (catalog image captions); A00176T180Y182 has human lung paraffin IHC and HeLa IF images (catalog image captions). A00176-2 has human breast cancer, mouse brain, and rat brain paraffin IHC images (catalog IHC captions); M00176 has a HeLa IF image and lists ICC as an application (catalog IF caption; catalog applications).
Which to pick: For tissue IHC, choose A00176-2 when its documented EDTA pH 8.0 retrieval and 2 μg/ml primary incubation suit the experiment (A00176-2 IHC caption). For IF/ICC, M00176 is a rabbit monoclonal with a HeLa IF image and listed ICC use (catalog clone and applications; M00176 IF caption). For IHC across human, mouse, and rat samples, A00176-2 has paraffin-section images in all three species; the captions do not report the fixative (A00176-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16539 (MK14_HUMAN, Mitogen-activated protein kinase 14).
- Human Protein Atlas. MAPK14 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MAPK14 subcellular location (ICC-IF): Mainly localized to the nuclear speckles. In addition localized to the cytosol..
- Human Protein Atlas. MAPK14 antibody validation summary (3 antibodies).
- Mapk14 is a Prognostic Biomarker and Correlates with the Clinicopathological Features and Immune Infiltration of Colorectal Cancer. Frontiers in cell and developmental biology 2022 — PMC8818961.
- Combined De-Repression of Chemoresistance Associated Mitogen-Activated Protein Kinase 14 and Activating Transcription Factor 2 by Loss of microRNA-622 in Hepatocellular Carcinoma. Cancers 2021 — PMC7967205.
- Bone morphogenetic protein 7 promotes resistance to immunotherapy. Nature communications 2020 — PMC7519103.
- Vascular smooth muscle-MAPK14 is required for neointimal hyperplasia by suppressing VSMC differentiation and inducing proliferation and inflammation. Redox biology 2019 — PMC6377391.
- PubMed PMID:7997261 — UniProt-cited evidence.
- PubMed PMID:7696354 — UniProt-cited evidence.
- PubMed PMID:7479834 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAPK14 (Mitogen-activated protein kinase 14). Expected localisation General cytoplasmic and nuclear staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet M00176-1). Validated IHC antibody: M00176-1 at 1:50 recommended; image 1ug/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded tissue sections; fixative not specified (datasheet M00176-1; sample unspecified); 2. Fixation: Image fixative and duration unreported (datasheet M00176-1); 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: Citrate pH 6 , 20 min (datasheet M00176-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00176-1); 8. Primary antibody: Rabbit monoclonal (clone EGC-13) anti-MAPK14, 1:50 recommended; image 1ug/ml (datasheet M00176-1); 9. Primary incubation: Overnight at 4 °C (datasheet M00176-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M00176-1); Caveat: Total MAPK14 staining does not establish activation (UniProt); Isoform / epitope: 5 isoforms; check epitope coverage for the chosen antibody (UniProt); Regulation: Cytokines and stress activate MAPK14 (UniProt). Evidence: UniProt Q16539, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nuclear speckles. In addition localized to…, Datasheet M00176-1, PMC8818961, PMC7967205. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAPK14/immunohistochemistry-ihc-guide.