This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MBNL1 / Muscleblind-like protein 1 · IHC design guide
Design Immunohistochemistry for MBNL1
Plan MBNL1 paraffin-section IHC around the nuclear staining seen in most tissues (HPA tissue IHC). Use HPA high-staining cells as positive controls and start the catalog antibody at 2–5 µg/mL (HPA tissue IHC; datasheet A02309-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MBNL1 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 tissue staining; cytoplasm also annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | Nuclear staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A02309-1) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+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 A02309-1) | |
| Caveat | Skeletal myocytes lack detectable IHC staining despite reported muscle expression (HPA tissue IHC; UniProt) | |
| Regulation | Higher during myoblast differentiation (UniProt) | |
| Isoform / epitope | Seven isoforms; antibody epitope coverage is unknown (UniProt) |
Section 1
Recommended MBNL1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A02309-1) with published MBNL1 staining protocols for transplanted tPVAT (PMC8698607), human brain and glioblastoma (PMC7642170), and stenotic intestine (PMC13297613).
| Sample | Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A02309-1) |
| Fixation | Image fixative and duration unreported (datasheet A02309-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A02309-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A02309-1) |
| Primary antibody | Rabbit anti-MBNL1, 2-5μg/ml (datasheet A02309-1) |
| Primary incubation | Overnight at 4 °C (datasheet A02309-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A02309-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MBNL1-positive staining in cells in molecular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: A02309-1); adjust staining conditions for the specimen and detection method.
Section 2
What Is the Expected MBNL1 Staining Pattern?
MBNL1 staining in paraffin sections is expected mainly in nuclei across many tissues (HPA tissue IHC: nuclear expression in most tissues). Strong examples include cerebellar molecular-layer cells, nasopharyngeal respiratory epithelium, skin arrector pili muscle cells and testicular spermatogonia (HPA tissue IHC: High). Cytoplasmic signal can also be biologically plausible (UniProt Q9NR56: nucleus and cytoplasm). MBNL1 has no transmembrane segment, so a membrane-restricted pattern is unexpected (UniProt Q9NR56 topology). HPA rates its tissue staining reliability Enhanced, with medium staining–RNA consistency (HPA tissue IHC).
| Distinct nuclear chromogen in the expected cells, with stronger staining in a documented High example (HPA tissue IHC: High). | This fits the reported nuclear pattern (HPA tissue IHC: nuclear expression in most tissues). Score the relevant cell population and compartment; compare intensity with controls processed in the same run (general IHC practice). |
| Staining is confined to cell borders or an extracellular structure, with little nuclear signal. | A membrane-restricted pattern conflicts with MBNL1’s lack of a transmembrane segment (UniProt Q9NR56 topology). Review morphology, background and antibody controls before interpreting it as target staining (general IHC practice). Cytoplasmic signal alone is less decisive because MBNL1 also localizes there (UniProt Q9NR56). |
| Prominent chromogen appears in an unexpected cell population, or persists when primary antibody is omitted. | Consider antibody cross-reactivity or endogenous chromogenic detection activity (general IHC practice). Check the omitted-primary control and tissue morphology before assigning MBNL1 expression; HPA documents cell-specific differences within tissues (HPA tissue IHC). |
| Diffuse color covers nuclei, cytoplasm and tissue spaces without clear cell boundaries. | This does not provide a reliable compartment call (general IHC practice). Review background controls, blocking, washing and chromogen development (general IHC practice); the reference pattern is chiefly nuclear (HPA tissue IHC). |
| No signal appears in a section containing a documented High cell population (HPA tissue IHC: High). | First treat the run as potentially uninformative: confirm that the relevant cells are present and that positive controls and detection worked (general IHC practice). Absence alone cannot distinguish low abundance from an assay failure (general IHC practice). |
💡Expected MBNL1 appearanceA convincing positive shows cell-associated, predominantly nuclear chromogen in a documented positive population, potentially strong in HPA High cells; isolated membrane-like or diffuse tissue-wide color is suspect (HPA tissue IHC: nuclear pattern and High examples; UniProt Q9NR56 topology; general IHC practice).
| Tissue and cell selection | Use the named cell population when judging a section: HPA reports High staining in cerebellar molecular-layer cells and nasopharyngeal respiratory epithelium, but Not detected in adipocytes and spleen red-pulp cells (HPA tissue IHC). A tissue name alone is an imprecise control designation (general IHC practice). |
| Tissue-source disagreement | UniProt describes high protein expression in cardiac and skeletal muscle, while HPA reports Low staining in cardiomyocytes and Not detected in skeletal myocytes (UniProt Q9NR56: tissue specificity; HPA tissue IHC). Treat those HPA cells cautiously as positive IHC controls; the records do not establish why they differ. |
| Antibody validation | HPA lists CAB016398 as IHC Enhanced and HPA035098 as IHC Supported (HPA antibodies). These are validation labels for the listed antibodies, not a guarantee that every section or detection run will reproduce a given intensity (HPA antibodies; general IHC practice). |
| Protein form and topology | UniProt lists seven MBNL1 isoforms, one annotated 1–388 chain, no signal peptide or propeptide, and no transmembrane segment (UniProt Q9NR56). Epitope coverage across isoforms is unspecified here; do not infer an isoform-specific stain from the observed compartment alone. |
| IF/ICC Q&A: Where should signal appear? | Mainly in nucleoplasm, with additional cytosol reported in ICC-IF (HPA subcellular: nucleoplasm Enhanced; cytosol Supported). UniProt also reports stress-granule localization upon stress (UniProt Q9NR56). These cell-image observations inform localization; they do not define a paraffin-section staining intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known High cells are present, but the section and run control are blank (HPA tissue IHC: High). | A shared staining or detection failure is plausible (general IHC practice). | Check the detection reagents and run control, then review the IHC-validated antibody’s stated IHC-P conditions before repeating the run (general IHC practice). |
| The run control stains, but the chosen test population is blank. | The selected cells may have little detectable staining: cardiomyocytes are Low and skeletal myocytes Not detected in HPA IHC (HPA tissue IHC). | Verify cell identity and compare with a documented High population on a suitable control section (HPA tissue IHC; general IHC practice). Do not call the entire assay failed from this population alone. |
| Brown color persists in the omitted-primary control. | Endogenous detection activity or another primary-independent background source may contribute (general chromogenic IHC practice). | Review the detection system’s background-blocking step and compare matched controls before scoring the test section (general IHC practice). |
| Staining is widespread and obscures nuclear boundaries. | Excess background or chromogen development can prevent compartment assessment (general IHC practice). | Review washing, blocking and development against the run controls; score MBNL1 only where cell boundaries and nuclear localization remain interpretable (general IHC practice; HPA tissue IHC: nuclear pattern). |
| Only cell borders stain while expected nuclei remain clear. | A border-only pattern does not match a protein without a transmembrane segment or HPA’s chiefly nuclear pattern (UniProt Q9NR56 topology; HPA tissue IHC). | Inspect morphology and background controls, then reassess antibody specificity before reporting the border signal as MBNL1 (general IHC practice). |
| Cytoplasmic staining accompanies nuclear staining. | Cytoplasmic localization is documented, although the HPA tissue IHC profile is chiefly nuclear (UniProt Q9NR56; HPA tissue IHC). | Record nuclear and cytoplasmic staining separately, compare with controls and avoid labeling cytoplasmic signal an artefact solely because it is extranuclear (general IHC practice; UniProt Q9NR56). |
Experiment evidence
IHC & IF images showing MBNL1 staining
A02309-1 · IHCIHC analysis of MBNL1 using anti-MBNL1 antibody (A02309-1). MBNL1 was detected in a paraffin-embedded section of human thyroid cancer tissue
A02309-1 · IHCIHC analysis of MBNL1 using anti-MBNL1 antibody (A02309-1). MBNL1 was detected in a paraffin-embedded section of human colorectal adenocarci
A02309-1 · IFIF analysis of MBNL1 using anti-MBNL1 antibody (A02309-1). MBNL1 was detected in an immunocytochemical section of A549 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000152601-MBNL1/tissue/cerebral+cortex#imid_16530915
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000152601-MBNL1/tissue/colon#imid_16530857
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000152601-MBNL1/tissue/liver#imid_16530842
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000152601-MBNL1/tissue/kidney#imid_16530843
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB016398; green = MBNL1, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB016398; green = MBNL1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MBNL1 IHC & IF
🧪Run skin first: arrector pili muscle cells should stain (HPA: High in arrector pili muscle cells). Use adipose tissue as the negative tissue (HPA: Not detected in adipocytes); on the skin slide, cells without specific staining should show counterstain without brown DAB signal, but adjacent cell types are not established as MBNL1-negative by these rows.
Positive control tissue: Cerebellum (Cells in molecular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MBNL1 in U-251MG, A-431, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a rabbit IgG isotype control matched to the primary antibody format; and, if available, MBNL1-knockout material as a biological specificity control (selected caption: rabbit primary and anti-rabbit secondary). Quench endogenous peroxidase for chromogenic IHC and distinguish skin pigment from DAB signal (selected caption: DAB detection; HPA: High in skin arrector pili muscle cells).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A02309-1 paraffin-section caption does not state the fixative (selected caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required across specimens (selected caption: EDTA retrieval). Frozen sections or IF are not established as easier by the supplied evidence; skin pigment can complicate DAB interpretation, while IF/ICC offers a way to assess the reported nucleoplasmic and cytosolic localization (HPA: nucleoplasm enhanced; cytosol supported).
HPA tissue IHC evidence for MBNL1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Nasopharynx | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Skin | Arrector pili muscle cells | High | Protein (IHC) | HPA → |
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced MBNL1 IHC Tips
Troubleshoot MBNL1 staining in paraffin sections by checking retrieval, compartment, controls and cell-specific scoring before interpreting changes in signal.
What retrieval should I try first for MBNL1 in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for this catalog antibody (datasheet A02309-1). The selected paraffin-section example used that retrieval before 2 μg/ml primary antibody overnight at 4°C, so it provides a practical starting condition (datasheet A02309-1). If staining is weak, vary heating and cooling conditions on matched sections while holding antibody concentration, detection and development constant (standard IHC practice). Compare nuclear signal in a suitable positive control with background on a no-primary control before accepting a stronger retrieval condition (HPA tissue IHC; standard IHC practice). Do not treat signal gained through damaged tissue or diffuse background as improved MBNL1 detection (standard IHC practice).
Could fixation explain weak or patchy MBNL1 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not report a fixative (datasheet A02309-1). Record fixation and processing history for each specimen, then compare similarly handled sections before changing the antibody concentration or retrieval conditions (standard IHC practice). In a controlled trial, keep EDTA pH 8.0 retrieval and the 2 μg/ml starting concentration fixed while comparing specimens with documented processing histories (datasheet A02309-1; standard IHC practice). Check morphology and nuclear counterstaining alongside MBNL1 signal, since poorly preserved or unevenly processed areas can distort a staining comparison (standard IHC practice). Do not infer a target-specific fixation effect from expression patterns or protein topology (HPA tissue IHC; UniProt Q9NR56).
How should I troubleshoot cytoplasmic staining when I expect nuclear MBNL1?
Expect predominantly nuclear staining across tissues, with nucleoplasm the main reported subcellular location; cytosolic signal is also reported (HPA tissue IHC; HPA subcellular). MBNL1 can occur in cytoplasm and stress granules under the described biological conditions, so cytoplasmic staining alone is not proof of artefact (UniProt Q9NR56). Inspect matched fields for a consistent intracellular pattern, preserved morphology and a clean no-primary control before assigning that signal to MBNL1 (standard IHC practice). If the pattern shifts between sections, repeat retrieval at EDTA pH 8.0 and compare staining under fixed detection conditions (datasheet A02309-1; standard IHC practice). Report nuclear and cytoplasmic scores separately rather than merging distinct compartments (standard IHC practice).
Can epitope accessibility or MBNL1 isoforms explain inconsistent staining?
MBNL1 has 7 listed isoforms, but the supplied antibody caption does not identify the recognized epitope or establish isoform coverage (UniProt Q9NR56; datasheet A02309-1). The protein has no annotated transmembrane segment, signal peptide or glycosylation sites, and a phosphothreonine is listed at residue 6 (UniProt Q9NR56). Those annotations do not establish how processing or retrieval affects this antibody’s epitope (UniProt Q9NR56; standard IHC practice). Compare matched sections with EDTA pH 8.0 retrieval and unchanged detection before attributing variable staining to epitope accessibility (datasheet A02309-1; standard IHC practice). Avoid assigning an isoform from staining intensity or compartment alone without independent epitope or isoform evidence (standard IHC practice).
How can I assess MBNL1 localisation by multiplex immunofluorescence?
Treat IF/ICC as a separate assay requiring its own antibody and fixation validation; the selected paraffin-section IHC caption supplies no IF conditions (datasheet A02309-1; standard IF practice). Multiplex MBNL1 with an independently validated marker of the expected cell type, and include single-stain controls to assess bleed-through (standard IF practice). Choose a spectrally separated, preferably far-red fluorophore when tissue autofluorescence obscures shorter wavelengths, and check an unstained section (standard IF practice). Because MBNL1 is nuclear and cytoplasmic and has no transmembrane segment, permeabilisation must permit access to the intracellular epitope; its precise location is unreported for this antibody (UniProt Q9NR56; datasheet A02309-1). Compare nuclear and cytoplasmic signal separately against matched negative controls (HPA subcellular; standard IF practice).
How do I reduce diffuse DAB background without losing MBNL1 signal?
Begin with the documented 10% goat serum block and 2 μg/ml primary antibody overnight at 4°C as reference conditions for this catalog antibody (datasheet A02309-1). Its selected paraffin-section example used a peroxidase-conjugated secondary for 30 minutes at 37°C and DAB development (datasheet A02309-1). Use a peroxidase block, adequate washing and a no-primary control to distinguish endogenous enzyme activity or nonspecific detection from antibody-dependent staining (standard IHC practice). If background persists, adjust one variable at a time and compare signal in the same compartment and cell type across matched sections (standard IHC practice). Keep DAB development comparable between test and control sections (standard IHC practice).
What should I score when comparing MBNL1 IHC across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear MBNL1 separately from cytoplasmic staining, since nuclear expression predominates while cytosolic localisation is also reported (HPA tissue IHC; HPA subcellular). For a defined compartment, record the percentage of positive cells and staining intensity, or calculate an H-score using consistent thresholds (standard IHC practice). Normalise positive-cell counts to the number of evaluable cells of that type, or report their density per mm² of evaluable tissue (standard IHC practice). Exclude folds, necrosis and damaged edges using the same rules for every section, and keep retrieval and DAB development comparable (standard IHC practice). Report compartment, denominator and scoring thresholds with each result (standard IHC practice).
Which staining patterns support genuine MBNL1 rather than artefact?
Give greatest weight to reproducible intracellular staining in intact cells, particularly nuclear staining, because the tissue profile is predominantly nuclear (HPA tissue IHC; standard IHC practice). Consider cytoplasmic signal in context: MBNL1 has reported cytoplasmic localisation, and stress-granule localisation occurs under stress (UniProt Q9NR56; HPA subcellular). Check whether the stained cells match the population being scored, since HPA reports differing levels among cell types, including high staining in cerebellar molecular-layer cells and undetected staining in adipocytes (HPA tissue IHC). Distrust signal restricted to section edges, necrotic regions or no-primary controls, and investigate endogenous peroxidase before calling DAB deposits positive (standard IHC practice). Do not infer a specific isoform or RNA-bound state from chromogenic staining alone (UniProt Q9NR56; standard IHC practice).
Boster reagents
Best MBNL1 / Muscleblind-like protein 1 IHC Antibodies
A02309-1 has IHC images from human paraffin sections and IF images from A549 cells and human colon cancer sections; the catalog lists Human, Monkey, Mouse and Rat reactivity (catalog image captions; catalog reactivity).
Real IHC data
Anti-MBNL1 Antibody ®
Cat # A02309-1
A02309-1 has IHC images from paraffin sections of human thyroid, colorectal, laryngeal and liver cancers (catalog IHC image captions). A02309-1 also has IF images from A549 cells and paraffin sections of human colon cancer (catalog IF image captions).
Which to pick: For chromogenic tissue IHC, choose A02309-1: its human paraffin section images use EDTA retrieval at pH 8.0, 2 μg/ml primary antibody and DAB detection; the fixative is unreported (catalog IHC image captions). For IF/ICC, A02309-1 has images from A549 cells and human colon cancer sections, with 5 μg/ml primary antibody (catalog IF image captions). For cross-species planning, the catalog lists Human, Monkey, Mouse and Rat reactivity for A02309-1, while the supplied IHC and IF images show human samples (catalog reactivity; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NR56 (MBNL1_HUMAN, Muscleblind-like protein 1).
- Human Protein Atlas. MBNL1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. MBNL1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. MBNL1 antibody validation summary (2 antibodies).
- Comparative Proteomic Analysis of tPVAT during Ang II Infusion. Biomedicines 2021 — PMC8698607.
- The Alternative Splicing Factor, MBNL1, Inhibits Glioblastoma Tumor Initiation and Progression by Reducing Hypoxia-Induced Stemness. Cancer research 2020 — PMC7642170.
- Muscleblind-like 1 is required for normal heart valve development in vivo. BMC developmental biology 2015 — PMC4608261.
- MBNL1 Promotes Intestinal Fibrosis via RAS-MAPK Pathway-Mediated Fibroblast Activation and Proliferation. Biomedicines 2026 — PMC13297613.
- PubMed PMID:9455477 — UniProt-cited evidence.
- PubMed PMID:10970838 — UniProt-cited evidence.
- PubMed PMID:11590133 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MBNL1 (Muscleblind-like protein 1). Expected localisation Nuclear tissue staining; cytoplasm also annotated (HPA tissue IHC; UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A02309-1). Validated IHC antibody: A02309-1 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in molecular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human thyroid cancer tissue; fixative not specified (datasheet A02309-1); 2. Fixation: Image fixative and duration unreported (datasheet A02309-1); 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: EDTA pH 8.0 (datasheet A02309-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A02309-1); 8. Primary antibody: Rabbit anti-MBNL1, 2-5μg/ml (datasheet A02309-1); 9. Primary incubation: Overnight at 4 °C (datasheet A02309-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A02309-1); Caveat: Skeletal myocytes lack detectable IHC staining despite reported muscle expression (HPA tissue IHC; UniProt); Isoform / epitope: Seven isoforms; antibody epitope coverage is unknown (UniProt); Regulation: Higher during myoblast differentiation (UniProt). Evidence: UniProt Q9NR56, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A02309-1, PMC8698607, PMC7642170. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MBNL1/immunohistochemistry-ihc-guide.