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MNAT1 / CDK-activating kinase assembly factor MAT1 · IHC design guide
Design Immunohistochemistry for MNAT1
Plan MNAT1 IHC-P staining around its nuclear pattern, using seminiferous duct cells as a positive reference (HPA tissue IHC). Start the catalog antibody at 0.5–1 μg/mL (datasheet PB9614) and assess staining by cell type.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MNAT1 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 | Nuclear staining in tissue cells (HPA tissue IHC) | |
| Staining pattern | Variable nuclear staining, prominent in seminiferous ducts (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB9614) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA have medium consistency (HPA tissue IHC) | |
| Regulation | Expression regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unspecified (UniProt) |
Section 1
Recommended MNAT1 IHC & IF Protocols
The catalog antibody protocol uses heat-mediated EDTA pH 8.0 retrieval (datasheet: PB9614). Published MNAT1 IHC examples cover colorectal cancer microarrays and paraffin-embedded laryngeal carcinoma sections (PMC6258412; PMC12274886).
| Sample | Paraffin-embedded mouse testis tissue; fixative not specified (datasheet PB9614) |
| Fixation | Image fixative and duration unreported (datasheet PB9614); 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 PB9614); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB9614) |
| Primary antibody | Rabbit anti-MNAT1, 0.5-1μg/ml (datasheet PB9614) |
| Primary incubation | Overnight at 4 °C (datasheet PB9614) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB9614) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MNAT1-positive staining in cells in seminiferous ducts of testis (HPA tissue IHC: High). HPA tissue profile: Nuclear expression at variable levels in several different cell types, most abundant in cells in seminiferous ducts, urinary bladder and gallbladder. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet: PB9614); the published examples do not specify retrieval conditions (PMC6258412; PMC12274886).
Section 2
What Is the Expected MNAT1 Staining Pattern?
MNAT1 should appear predominantly in nuclei, with the strongest reported IHC staining in cells of testis seminiferous ducts and variable staining in other cell types (UniProt P51948: nucleus; HPA tissue IHC: High in cells in seminiferous ducts). It has no transmembrane segment (UniProt P51948 topology). HPA rates its tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA tissue IHC reliability).
| Clear nuclear staining in cells of seminiferous ducts; weaker nuclear staining in some other tissues. | This matches HPA’s High testis signal and variable nuclear tissue profile (HPA tissue IHC). Compare the named cell type and its nuclei, since an entire tissue section is not uniformly positive (HPA tissue IHC). |
| Predominantly cytoplasmic or membrane-like staining, with little nuclear signal. | Treat this as a localization mismatch requiring investigation: MNAT1 is nuclear and lacks a transmembrane segment (UniProt P51948: subcellular location and topology). HPA also reports nucleoplasmic ICC-IF staining (HPA subcellular). |
| Strong staining in skeletal myocytes, adipocytes, or splenic red-pulp cells. | These cell types were Not detected by HPA tissue IHC (HPA tissue IHC). Consider cross-reactivity or endogenous chromogenic detection activity (general IHC practice); first verify which cells actually carry the stain. |
| A broad haze covers nuclei, cytoplasm, and surrounding tissue. | A haze without cell-specific nuclear contrast is difficult to score as MNAT1 (UniProt P51948: nucleus; HPA tissue IHC: nuclear profile). Review background controls, blocking, and detection conditions (general IHC practice). |
| No staining in cells of testis seminiferous ducts. | The expected High positive reference is absent (HPA tissue IHC). Before calling a sample negative, check section quality and control performance, then review the antibody’s IHC conditions and detection reagents (general IHC practice). |
💡Expected MNAT1 appearanceCall a positive result when staining is predominantly nuclear in the relevant cells, strongest in cells of testis seminiferous ducts; diffuse or chiefly cytoplasmic staining is suspect (HPA tissue IHC: High in seminiferous ducts and nuclear profile; UniProt P51948: nucleus).
| Subcellular location | Nuclear staining is the principal scoring feature (UniProt P51948: nucleus; HPA tissue IHC: nuclear profile). Membrane-like staining conflicts with the absence of a transmembrane segment (UniProt P51948 topology). |
| Cell-type variation | HPA reports Medium staining in lung macrophages and several glandular cell types, but Not detected in skeletal myocytes and splenic red-pulp cells (HPA tissue IHC). Judge staining by the identified cells. |
| Strength of tissue evidence | HPA calls the tissue IHC profile Approved while describing only medium consistency with RNA expression (HPA tissue IHC reliability). Its observed cell-level contrasts guide interpretation; they do not guarantee every specimen’s result. |
| Antibody-specific validation | HPA lists HPA000701 and CAB004495 as IHC Approved (HPA antibodies). That status belongs to those antibodies; it does not establish equivalent staining or an IHC dilution for another antibody. |
| Isoforms and processing | UniProt lists 2 isoforms and a single chain spanning residues 1–309, without a signal peptide or propeptide (UniProt P51948). The supplied record gives no antibody epitope, so isoform-specific detection cannot be predicted. |
| IF/ICC reference | HPA reports enhanced nucleoplasmic ICC-IF localization with HPA001154 (HPA subcellular; HPA antibodies). This supports the compartment expectation, but ICC-IF validation does not establish an IHC staining intensity or protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive testis section has no nuclear signal. | A failed assay or poorly performing control is possible; HPA reports High staining in cells of seminiferous ducts (HPA tissue IHC). | Confirm those cells are present, inspect the positive control, and check the antibody’s IHC instructions, retrieval step, and detection reagents (general IHC practice). |
| Most of the section shows uniform brown haze. | Background from detection chemistry or insufficient blocking can obscure cell-specific contrast (general IHC practice). | Compare a no-primary control, review blocking and wash steps, and adjust detection conditions before scoring nuclear MNAT1 (general IHC practice; UniProt P51948: nucleus). |
| Cytoplasmic or membrane-like staining dominates. | The compartment conflicts with UniProt’s nuclear assignment and absence of a transmembrane segment (UniProt P51948: subcellular location and topology). | Compare a known-positive section and inspect nuclear counterstain alignment; review primary-antibody specificity and detection background (general IHC practice). |
| An HPA Not detected cell type stains strongly. | For example, HPA reports skeletal myocytes as Not detected; cross-reactivity or endogenous detection activity may explain unexpected chromogen (HPA tissue IHC; general IHC practice). | Identify the stained cells, examine a no-primary control, and confirm the pattern with an independently validated antibody if available (general IHC practice). |
| Signal varies between cells or tissue regions. | MNAT1 staining varies by cell type in HPA tissue IHC; HPA also reports only medium consistency with RNA expression (HPA tissue IHC reliability). | Score the intended cell population separately and compare it with the expected cell-level pattern before treating variation as assay failure (HPA tissue IHC). |
| Can IF/ICC settle an uncertain IHC compartment call? | HPA’s ICC-IF result is nucleoplasmic and enhanced, while its tissue IHC profile is nuclear and Approved (HPA subcellular; HPA antibodies; HPA tissue IHC). | Use the ICC-IF localization as a compartment cross-check; follow the separate IF/ICC guide for that application. Do not infer an IHC protocol from ICC-IF validation. |
Experiment evidence
IHC & IF images showing MNAT1 staining
PB9614 · IHCIHC analysis of MNAT1 using anti-MNAT1 antibody (PB9615). MNAT1 was detected in a paraffin-embedded section of mouse testis tissue. Heat med
PB9614 · IHCIHC analysis of MNAT1 using anti-MNAT1 antibody (PB9615). MNAT1 was detected in a paraffin-embedded section of rat testis tissue. Heat media
PB9614 · IFIF analysis of MNAT1 using anti-MNAT1 antibody (PB9614). MNAT1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retr
A06523 · IFIF analysis of MNAT1 using anti-MNAT1 antibody (A06523) and anti-Beta Tubulin antibody (M01857-3). MNAT1 was detected in immunocytochemical
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000020426-MNAT1/tissue/cerebral+cortex#imid_212122
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000020426-MNAT1/tissue/colon#imid_212040
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000020426-MNAT1/tissue/liver#imid_212041
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000020426-MNAT1/tissue/kidney#imid_212050
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001154; green = MNAT1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA001154; green = MNAT1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MNAT1 IHC & IF
🧪Run testis first and look for nuclear staining in cells of the seminiferous ducts (HPA: High in cells in seminiferous ducts; UniProt P51948: nucleus). Use heart muscle cardiomyocytes as the tissue negative (HPA: Not detected in cardiomyocytes); on the testis slide, treat unstained nuclei as internal negatives only where they are actually observed, since the supplied HPA row does not identify a specific negative cell type there.
Positive control tissue: Testis (Cells in seminiferous ducts, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MNAT1 in A-431, U-251MG, U2OS, U2OS, siRNA 1 (10x), U2OS, siRNA 1 (40x), U2OS, scrambled (10x), U2OS, scrambled (40x), with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an immunoglobulin-class-matched isotype control matched to the primary host species (caption: rabbit primary); use MNAT1-knockout material as a biological negative if available (standard IHC practice). Quench endogenous peroxidase and check for endogenous biotin background in testis because the caption uses biotin–streptavidin detection with DAB (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected tissue-IHC caption does not state a fixative (caption: fixative not stated). The mouse-testis paraffin-section caption reports heat retrieval in EDTA at pH 8.0, but it does not establish that retrieval is required for every specimen or antibody (caption: EDTA heat retrieval). Whether frozen sections or IF are easier is unreported; the caption’s biotin-based detection makes endogenous biotin a background concern in testis, while ICC-IF images place MNAT1 in the nucleoplasm (caption: SABC detection; HPA subcellular: nucleoplasm).
HPA tissue IHC evidence for MNAT1
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 |
|---|---|---|---|---|
| Testis | Cells in seminiferous ducts | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Parathyroid gland | 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 → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MNAT1 IHC Tips
Troubleshoot MNAT1 staining in paraffin sections by checking retrieval, nuclear localisation, tissue controls and scoring (UniProt P51948; HPA tissue IHC).
What should I change when MNAT1 nuclear staining is weak after retrieval?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PB9614). Check that sections were fully deparaffinised, remained covered by retrieval solution and cooled consistently before staining (standard IHC practice). Compare a weak section with a concurrently processed testis control, scoring nuclei in seminiferous ducts rather than diffuse tissue colour (HPA: High in cells in seminiferous ducts; standard IHC practice). If staining remains weak, test a different retrieval condition on adjacent sections as a fallback and compare signal with tissue damage and background (standard IHC practice). Record the retrieval method with each image because a darker DAB reaction alone does not establish improved MNAT1 detection (standard IHC practice).
How can I troubleshoot suspected fixation effects on MNAT1 staining?
MNAT1-specific sensitivity to fixation is unknown from the supplied evidence, so do not assign weak staining to a particular fixative or fixation duration (supplied evidence: no target-specific fixation data). The selected caption describes a paraffin section but does not state its fixative; it also names antibody PB9615 while the selected SKU is PB9614 (selected tissue-IHC caption). Record the actual fixative, processing history and section age, then compare sections processed together before changing retrieval or antibody conditions (standard IHC practice). Include a testis control and evaluate staining in cells of seminiferous ducts, where nuclear signal is reported as high (HPA tissue IHC).
Should MNAT1 staining appear in nuclei or cytoplasm?
Evaluate nuclear DAB staining first: MNAT1 is reported in the nucleus, with enhanced nucleoplasmic localisation in ICC/IF (UniProt P51948 subcellular; HPA subcellular). Compare the brown reaction product with a nuclear counterstain and inspect individual cells at a magnification that resolves nuclear boundaries (standard IHC practice). Strong cytoplasmic or surface staining without corresponding nuclear staining warrants checks of background, section folds and the secondary detection system before interpretation (UniProt P51948 subcellular; standard IHC practice). MNAT1 has no transmembrane segment, so a crisp membrane-only pattern is inconsistent with the supplied topology and should prompt review of staining specificity (UniProt P51948 topology).
Could isoforms or epitope accessibility explain inconsistent MNAT1 IHC?
MNAT1 has 2 listed isoforms, but the supplied evidence does not identify which isoform or epitope the IHC-validated antibody detects (UniProt P51948 isoforms; supplied antibody evidence). Do not interpret a weak cell population as isoform-specific without an antibody epitope map and independent validation (standard IHC practice). The protein contains a UIM at residues 142–161 and reported phosphorylation sites at 51 and 279; these annotations alone do not predict retrieval behaviour (UniProt P51948 domains and modified residues). If staining varies between runs, compare matched sections under the same retrieval and detection conditions before proposing epitope masking (standard IHC practice).
How should I check MNAT1 localisation by multiplex IF?
Use IF/ICC as a separate localisation check and assess nucleoplasmic signal, the enhanced location reported for MNAT1 (HPA subcellular). For tissue multiplexing, pair MNAT1 with a validated marker identifying the expected cell population, such as cells in seminiferous ducts, and inspect marker overlap at single-cell resolution (HPA: High in cells in seminiferous ducts; standard IF practice). Choose fluorophores and imaging channels after measuring tissue autofluorescence with appropriate controls (standard IF practice). Because MNAT1 is nuclear and has no transmembrane segment, use permeabilisation appropriate for access to an intracellular epitope, then compare nuclear signal with a DNA counterstain (UniProt P51948 subcellular and topology; standard IF practice).
What causes widespread brown staining that obscures MNAT1-positive nuclei?
First compare the stained section with a no-primary control to identify colour generated by secondary reagents, endogenous enzyme activity or the detection system (standard IHC practice). For a peroxidase/DAB workflow, check the peroxidase block and review whether DAB development is producing diffuse deposits (standard IHC practice). Examine section edges, folds and damaged areas separately, because local reagent pooling or tissue damage can mimic increased nuclear staining (standard IHC practice). The selected image caption describes goat-serum blocking, a biotinylated secondary antibody and DAB, but names PB9615 rather than the selected PB9614 SKU; treat those details as caption evidence for PB9615 only (selected tissue-IHC caption).
How should I score MNAT1 IHC across heterogeneous tissue? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell compartment and regions before scoring, then count nuclear-positive cells within each annotated cell population rather than averaging whole-section DAB colour (UniProt P51948 subcellular; standard IHC practice). Report either percentage of positive nuclei or an H-score combining staining intensity with the percentage at each intensity, using one scoring rule across sections (standard IHC practice). Normalise counts to the number of eligible nuclei in each region; for spatial counts, report positive nuclei per mm² of viable, annotated tissue (standard IHC practice). Keep cell types separate because HPA reports high staining in cells in seminiferous ducts and no detection in adipocytes (HPA tissue IHC).
How do I distinguish genuine MNAT1 signal from staining artefact?
A plausible positive is predominantly nuclear and occurs in an expected cell population, particularly cells in seminiferous ducts, where HPA reports high staining (UniProt P51948 subcellular; HPA tissue IHC). Compare that pattern with adipocytes or cardiomyocytes, reported as not detected, while recognising that HPA rates tissue staining as Approved with medium RNA agreement (HPA tissue IHC). Treat membrane-only staining, edge-heavy colour, necrotic areas and signal shared by a no-primary control as reasons to investigate artefact (UniProt P51948 topology; standard IHC practice). Check the peroxidase block if diffuse DAB persists, and avoid calling a section positive solely because its overall colour is darker (standard IHC practice).
Boster reagents
Best MNAT1 / CDK-activating kinase assembly factor MAT1 IHC Antibodies
The catalog lists anti-MNAT1 antibodies for IHC and IF/ICC in human, mouse and rat (catalog: PB9614, A06523); the supplied figures show paraffin-section IHC and cell IF (image captions).
Real IHC data
Anti-MNAT1 Antibody ®
Cat # PB9614
Real IF data
Anti-MNAT1 Antibody ®
Cat # A06523
PB9614 is listed for IHC and IF/ICC in human, mouse and rat; its associated IHC captions show paraffin sections of human, mouse and rat testis but identify the antibody as PB9615 (catalog: applications/reactivity; IHC captions). A06523 is listed for IF/ICC in human, mouse and rat, with a HELA cell IF figure and no listed IHC application (catalog: applications/reactivity; IF caption).
Which to pick: For paraffin-section IHC, choose PB9614 based on its listed application and 0.5–1 μg/ml IHC range, while checking the PB9614/PB9615 caption discrepancy before relying on the images (catalog: PB9614 applications/dilution; IHC captions). The captions report paraffin sections and EDTA retrieval at pH 8.0, but do not report the fixative (IHC captions). For IF/ICC, A06523 has a HELA cell figure at 5 μg/ml; both SKUs list human, mouse and rat reactivity, while the supplied IF figures document cells only (catalog: applications/reactivity; IF captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P51948 (MAT1_HUMAN, CDK-activating kinase assembly factor MAT1).
- Human Protein Atlas. MNAT1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MNAT1 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. MNAT1 antibody validation summary (3 antibodies).
- MNAT1 is overexpressed in colorectal cancer and mediates p53 ubiquitin-degradation to promote colorectal cancer malignance. Journal of experimental & clinical cancer research : CR 2018 — PMC6258412.
- Mitochondrial apoptosis induced by MNAT1 in laryngeal squamous cell carcinoma cells reverses drug resistance. Translational oncology 2025 — PMC12274886.
- The regulatory effect of has-circ-0001146/miR-26a-5p/MNAT1 network on the proliferation and invasion of osteosarcoma. Bioscience reports 2020 — PMC7284318.
- MNAT1 promotes proliferation and the chemo-resistance of osteosarcoma cell to cisplatin through regulating PI3K/Akt/mTOR pathway. BMC cancer 2020 — PMC7713032.
- PubMed PMID:8521818 — UniProt-cited evidence.
- PubMed PMID:8521393 — UniProt-cited evidence.
- PubMed PMID:12508121 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MNAT1 (CDK-activating kinase assembly factor MAT1). Expected localisation Nuclear staining in tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB9614). Validated IHC antibody: PB9614 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Cells in seminiferous ducts); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse testis tissue; fixative not specified (datasheet PB9614); 2. Fixation: Image fixative and duration unreported (datasheet PB9614); 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 PB9614); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB9614); 8. Primary antibody: Rabbit anti-MNAT1, 0.5-1μg/ml (datasheet PB9614); 9. Primary incubation: Overnight at 4 °C (datasheet PB9614). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining and RNA have medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unspecified (UniProt); Regulation: Expression regulation is unreported (UniProt). Evidence: UniProt P51948, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet PB9614, PMC6258412, PMC12274886. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MNAT1/immunohistochemistry-ihc-guide.