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MKNK1 / MAP kinase-interacting serine/threonine-protein kinase 1 · IHC design guide
Design Immunohistochemistry for MKNK1
Plan MKNK1 staining in paraffin sections using its general cytoplasmic tissue pattern (HPA tissue IHC). Compare staining across consistently fixed sections and account for possible isoform-dependent localisation (standard IHC practice; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MKNK1 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); nuclear location annotated (UniProt) | |
| Staining pattern | General cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Isoform-dependent localisation may alter staining (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt; datasheet) |
Section 1
Recommended MKNK1 IHC & IF Protocols
The catalog antibody protocol is followed by published MKNK1 IHC protocols for tissue sections (PMC9957794) and mouse primary tumors (PMC11397505).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A03783-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-MKNK1, 1:50-1:200 (datasheet A03783-2) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MKNK1-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 Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval setting; UniProt Q9BUB5: nucleus).
Section 2
What Is the Expected MKNK1 Staining Pattern?
MKNK1 is reported in the cytoplasm and nucleus and has no transmembrane segment (UniProt Q9BUB5). In tissue IHC, expect a general cytoplasmic pattern, including strong staining in the cell populations listed by HPA (HPA: tissue IHC). HPA rates the tissue profile Approved, with medium consistency between antibody staining and RNA expression; its annotation is based on CAB037310 (HPA: tissue IHC).
| Cytoplasmic staining in appendix or colon glandular cells, or bronchial respiratory epithelial cells. | This fits the general tissue pattern and named high staining populations (HPA: tissue IHC). Compare intensity with neighboring cells on the same section; HPA levels describe observed staining, not a numerical scoring cutoff. |
| Predominantly nuclear staining, or a nuclear signal alongside cytoplasmic staining. | Check the assay context before rejecting it: UniProt lists both compartments, while HPA tissue IHC describes general cytoplasmic expression (UniProt Q9BUB5; HPA: tissue IHC). HPA reports supported nucleoplasmic localization by ICC-IF and cautions that localization may depend on isoform (HPA: subcellular; HPA: tissue IHC). |
| A crisp membrane rim or extracellular deposit dominates the signal. | This is outside the reported cytoplasmic and nuclear locations and is unexplained by MKNK1 topology (UniProt Q9BUB5). Treat it as suspect staining; check the detection control and compare a second IHC-validated antibody before assigning it to MKNK1 (standard IHC practice). |
| Strong staining appears in an unexpected cell population while expected cells remain weak. | Possible causes include cross-reactivity or endogenous detection activity (standard IHC practice). HPA lists low staining in spleen red pulp cells, but its tissue list is not an exhaustive map of negative cells (HPA: tissue IHC). Confirm the cell identity and staining controls before interpreting the difference. |
| No visible signal in a selected high staining reference tissue. | For example, HPA reports high staining in appendix glandular cells and bone marrow hematopoietic cells (HPA: tissue IHC). A blank result there warrants checks of section quality, antibody dilution, retrieval, and detection reagents (standard IHC practice); it does not alone establish absent MKNK1. |
💡Expected MKNK1 appearanceCall a result consistent with MKNK1 when cytoplasmic staining is clear in an HPA high staining cell population, such as appendix glandular cells; a dominant membrane rim is suspect (HPA: tissue IHC; UniProt Q9BUB5 topology).
| Tissue and cell selection | HPA reports high staining in specified glandular, epithelial, hematopoietic, neuronal, Purkinje, and endothelial populations; spleen red pulp cells are low (HPA: tissue IHC). Select a named high staining population as a reference without treating other cells as established negatives. |
| Antibody validation | CAB027355 and CAB037310 have Approved IHC status; HPA's tissue profile is based on CAB037310 and has medium staining-to-RNA consistency (HPA: antibodies; HPA: tissue IHC). Interpret discrepancies in light of that qualification. |
| Isoforms and compartment | UniProt records three isoforms and both cytoplasmic and nuclear locations (UniProt Q9BUB5). HPA notes possible isoform-dependent localization, so an IHC–ICC compartment difference needs antibody and assay context (HPA: tissue IHC). |
| IF/ICC Q: What should appear? | A: HPA reports supported nucleoplasmic localization in ICC-IF, with images from HaCaT, RT-4, and U2OS (HPA: subcellular). That observation does not replace the general cytoplasmic tissue IHC pattern (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| High staining reference cells are blank. | The HPA reference predicts staining, but a negative slide can reflect an assay failure (HPA: tissue IHC; standard IHC practice). | Check section integrity and controls, then review retrieval, catalog antibody dilution, and detection steps; no target-specific retrieval condition is supplied (standard IHC practice). |
| Brown signal covers most tissue structures without cell boundaries. | Diffuse background can arise from detection reagents, blocking, or excess primary antibody (standard IHC practice). | Examine a no-primary control, review blocking and washes, and titrate the primary antibody while retaining a high staining reference section (standard IHC practice; HPA: tissue IHC). |
| Signal follows vessel or cell membranes as a sharp outline. | A membrane-dominant pattern is inconsistent with the listed intracellular locations and lack of a transmembrane segment (UniProt Q9BUB5). | Review morphology and detection controls; repeat with an independently IHC-validated antibody if available (standard IHC practice; HPA: antibodies). |
| Nuclear signal exceeds cytoplasmic signal. | The tissue profile is generally cytoplasmic, while supported ICC-IF localization is nucleoplasmic; HPA flags possible isoform dependence (HPA: tissue IHC; HPA: subcellular). | Record both compartments and the antibody used, then compare with a named high staining tissue population before calling the signal nonspecific (HPA: tissue IHC; standard IHC practice). |
| Unexpected cells stain more strongly than nearby expected cells. | Cell identification, cross-reactivity, or endogenous detection activity may explain the contrast (standard IHC practice). HPA's named levels do not define every cell as positive or negative (HPA: tissue IHC). | Recheck cell morphology and a no-primary control, and compare an independent IHC-validated antibody where available (standard IHC practice; HPA: antibodies). |
| IHC appears cytoplasmic but IF/ICC appears nucleoplasmic. | These are the respective reported patterns, and localization may be isoform dependent (HPA: tissue IHC; HPA: subcellular). | Keep the readouts assay-specific; record antibody identity and compartment scoring, and consult the separate IF/ICC guide for its workflow (HPA: antibodies; standard IHC/IF practice). |
Experiment evidence
IHC & IF images showing MKNK1 staining
A03783-2 · IHCImmunohistochemistry (IHC) analyzes of Mnk1 (I136) pAb in brain tissue of human
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000079277-MKNK1/tissue/cerebral+cortex#imid_9688156
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000079277-MKNK1/tissue/colon#imid_9688074
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000079277-MKNK1/tissue/liver#imid_9688075
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000079277-MKNK1/tissue/kidney#imid_9688068
HPA · ICC-IF HaCaTHuman Protein Atlas immunofluorescence, HaCaT, antibody HPA071293; green = MKNK1, blue = nucleus, red = microtubules.
HPA · ICC-IF HaCaTHuman Protein Atlas immunofluorescence, HaCaT, antibody HPA071293; green = MKNK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MKNK1 IHC & IF
🧪Run appendix first and assess staining in its glandular cells (HPA: High in appendix glandular cells). HPA lists no negative tissue and detects MKNK1 in all 45 scored tissues, so no-primary and isotype controls carry the negative comparison; any unstained cells on the appendix slide should show only background signal and should not be treated as validated MKNK1-negative cells (HPA: no negative tissue listed).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: None in HPA: MKNK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MKNK1 in HaCaT, RT-4, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Process no-primary (secondary-only) and host-species-matched isotype controls alongside the positive section; use MKNK1 knockout material, or immunizing-peptide competition when the peptide is known, to test antibody specificity (standard IHC practice). For chromogenic detection, quench endogenous peroxidase and check endogenous biotin if using a biotin-based system; for IF, assess tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A03783-2 tissue-IHC caption does not state a fixative (selected-SKU caption: fixative not stated). Retrieval dependency is unreported; optimize antigen retrieval for paraffin sections empirically (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier, although HPA reports nucleoplasmic ICC-IF localization in HaCaT, RT-4 and U2OS cells (HPA: subcellular); assess appendix luminal deposits separately from cell-associated staining (standard IHC practice).
HPA tissue IHC evidence for MKNK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. External characterization data suggest that subcellular localization is isoform dependent. Annotated expression profile is based on CAB037310.). 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 → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: MKNK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced MKNK1 IHC Tips
Troubleshoot MKNK1 staining in chromogenic IHC of paraffin sections, with one entry on translating the analysis to IF.
How should I retrieve MKNK1 when staining is weak?
Start with Tris-EDTA at pH 9.0 using heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval rule). Keep section thickness, heating vessel, and cooling time consistent across the run, then compare signal in the same cell population with a no-primary control (standard IHC practice). The selected catalog image shows MKNK1 staining in human brain, but its caption reports no fixative or retrieval method, so it cannot establish retrieval sensitivity for SKU A03783-2 (catalog caption: A03783-2). If staining remains weak, test a milder retrieval condition on adjacent sections while tracking tissue damage and background, and retain the condition that best preserves interpretable cellular staining (standard IHC practice).
Could fixation be masking MKNK1 in my sections?
Target-specific fixation sensitivity is unknown: the selected human-brain IHC caption does not state a fixative, and the supplied record gives no fixation comparison (catalog caption: A03783-2). Document the fixative, fixation duration, section thickness, and storage history for each specimen before attributing a weak result to MKNK1 itself (standard IHC practice). Compare adjacent sections using the page's Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 minutes, keeping detection and exposure to chromogen consistent (page retrieval rule; standard IHC practice). If tissue preservation or staining varies with processing, validate the chosen fixation workflow on matched material and report that result as a local observation (standard IHC practice).
Should MKNK1 appear in nuclei or cytoplasm?
Assess both compartments: UniProt lists MKNK1 in cytoplasm and nucleus, while HPA reports general cytoplasmic tissue staining and supported nucleoplasmic localisation in its subcellular data (UniProt Q9BUB5 subcellular; HPA tissue IHC; HPA subcellular). Score nuclear and cytoplasmic signal separately after a light hematoxylin counterstain, using the same chromogen development time for compared sections (standard IHC practice). HPA also notes evidence that localisation depends on isoform, so a compartment difference alone does not establish an artefact (HPA tissue IHC). If signal appears exclusively along cell membranes, examine the no-primary control and section morphology before assigning it to MKNK1, which has no transmembrane segment (UniProt Q9BUB5 topology; standard IHC practice).
Could isoforms or phosphorylation explain inconsistent staining?
MKNK1 has 3 recorded isoforms and a kinase domain spanning residues 49–374, so antibody epitope placement matters when comparing specimens or reagents (UniProt Q9BUB5 isoforms and domains). Check the catalog antibody's stated immunogen and whether it distinguishes isoforms before interpreting a change in staining as a change in total MKNK1 (standard antibody validation practice). Recorded modified residues include Ser39, Ser221, Ser226, Thr250, and Thr255, but the supplied evidence does not show that any of them changes this antibody's binding (UniProt Q9BUB5 modified residues). Compare adjacent sections under identical 20-minute retrieval and chromogen conditions, then describe unresolved differences as staining differences rather than isoform- or phosphorylation-specific results (page retrieval rule; standard IHC practice).
How can I check MKNK1 localisation by multiplex IF?
For a secondary IF assay, pair MKNK1 with a validated marker for the cell population being examined, such as a neuronal marker in caudate, where HPA reports high staining in neuronal cells (HPA: caudate neuronal cells High; standard IF practice). Choose fluorophores after imaging an unstained section for tissue autofluorescence, and place the weaker signal in a channel with good separation from that background (standard IF practice). MKNK1 is intracellular and has no transmembrane segment, so use a controlled permeabilisation step for access to cytoplasmic and nuclear epitopes, then check whether it alters morphology (UniProt Q9BUB5 topology and subcellular; standard IF practice). Test secondary-only and single-colour controls before interpreting nuclear overlap, because HPA reports supported nucleoplasmic localisation but does not validate this catalog antibody for IF (HPA subcellular; standard IF practice).
What should I check when chromogenic background obscures MKNK1?
Run a no-primary control beside the MKNK1 section to separate primary-dependent signal from endogenous enzyme activity and nonspecific detection staining (standard IHC practice). For a peroxidase and DAB workflow, include a peroxidase block, then inspect whether residual colour follows blood-rich areas or damaged tissue rather than intact cells (standard IHC practice). Titrate the primary antibody and shorten chromogen development in matched sections while keeping retrieval at Tris-EDTA pH 9.0 and 95–98 °C for 20 minutes constant (page retrieval rule; standard IHC practice). Preserve enough counterstain to identify cell borders, since HPA describes general cytoplasmic tissue expression and background can otherwise resemble diffuse MKNK1 signal (HPA tissue IHC; standard IHC practice).
How should I score MKNK1 staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because HPA reports general cytoplasmic tissue expression while its subcellular data support nucleoplasmic localisation (HPA tissue IHC; HPA subcellular). Record 0–3 intensity and the percentage of cells at each intensity, then calculate an H-score as the sum of intensity multiplied by its percentage, with a possible range of 0–300 (standard IHC scoring practice). Alternatively, report percentage positive cells or positive-cell density per mm² within a predefined region, using one threshold across compared sections (standard IHC practice). Normalise counts to the number of eligible cells or evaluable tissue area, exclude necrotic and folded regions, and keep retrieval and imaging settings consistent (standard IHC practice).
When is apparent MKNK1 positivity likely to be artefact?
A plausible result should resolve to intact cells in cytoplasm or nucleus, consistent with UniProt localisation and HPA's tissue and subcellular observations (UniProt Q9BUB5 subcellular; HPA tissue IHC; HPA subcellular). Compare the labelled cell type with the section anatomy: HPA reports high staining in caudate neuronal cells and cerebral-cortex endothelial cells, among other populations (HPA: caudate neuronal cells High; HPA: cerebral cortex endothelial cells High). Treat sharp section-edge staining, necrotic deposits, and signal in a no-primary control as reasons to investigate processing or endogenous detection activity (standard IHC practice). Check adjacent intact fields and repeat with consistent 20-minute retrieval before claiming a biological compartment shift, especially because HPA notes isoform-dependent localisation evidence (page retrieval rule; HPA tissue IHC; standard IHC practice).
Boster reagents
Best MKNK1 / MAP kinase-interacting serine/threonine-protein kinase 1 IHC Antibodies
The catalog lists one IHC antibody with a human brain tissue image and stated human, mouse, and rat reactivity (catalog: A03783-2; IHC image caption). No IF image is supplied (catalog: A03783-2).
Real IHC data
Anti-Mnk1 (I136) MKNK1 Antibody
Cat # A03783-2
A03783-2 is the only SKU that will render; its IHC image shows human brain tissue (IHC image caption: A03783-2). The catalog lists IHC and WB applications and human, mouse, and rat reactivity (catalog: A03783-2).
Which to pick: For tissue IHC, choose A03783-2: it lists IHC and has a human brain tissue image (catalog and IHC image caption: A03783-2). For cross-species work, A03783-2 lists human, mouse, and rat reactivity, though its supplied IHC image shows only human tissue (catalog and IHC image caption: A03783-2). No IF/ICC application or image is listed, so an IF/ICC choice is unsupported; the IHC caption does not report section processing or fixative (catalog and IHC image caption: A03783-2).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BUB5 (MKNK1_HUMAN, MAP kinase-interacting serine/threonine-protein kinase 1).
- Human Protein Atlas. MKNK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MKNK1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. MKNK1 antibody validation summary (3 antibodies).
- Identification of MKNK1 and TOP3A as ovarian endometriosis risk-associated genes using integrative genomic analyses and functional experiments. Computational and structural biotechnology journal 2023 — PMC9957794.
- Blocking tumor-intrinsic MNK1 kinase restricts metabolic adaptation and diminishes liver metastasis. Science advances 2024 — PMC11397505.
- PubMed PMID:9155018 — UniProt-cited evidence.
- PubMed PMID:11463832 — UniProt-cited evidence.
- PubMed PMID:15350534 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MKNK1 (MAP kinase-interacting serine/threonine-protein kinase 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location annotated (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A03783-2 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A03783-2); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-MKNK1, 1:50-1:200 (datasheet A03783-2); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Isoform-dependent localisation may alter staining (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt; datasheet); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt Q9BUB5, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A03783-2, PMC9957794, PMC11397505. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MKNK1/immunohistochemistry-ihc-guide.