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MEF2B / Myocyte-specific enhancer factor 2B · IHC design guide
Design Immunohistochemistry for MEF2B
Plan MEF2B IHC in paraffin sections around selective nuclear staining in germinal center lymphocytes of tonsil and lymph node (HPA tissue IHC). The guide covers fixation consistency, interpretation of the staining pattern, and the cross-gene detection caveat (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MEF2B 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 in germinal center lymphocytes (HPA tissue IHC) | |
| Staining pattern | Selective nuclear staining in tonsil and lymph node germinal centers (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05743-1) | |
| Positive control | Lymph node+1 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 A05743-1) | |
| Caveat | Antibody may detect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Lymphoid tissue enriched; intensity may vary by cell population (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope differences are unspecified (UniProt) |
Section 1
Recommended MEF2B IHC & IF Protocols
The catalog antibody’s IHC-P protocol and one published MEF2B tumor-core IHC method provide starting conditions (PMC11143196).
| Sample | Paraffin-embedded human hodgkin lymphoma tissue; fixative not specified (datasheet A05743-1) |
| Fixation | Image fixative and duration unreported (datasheet A05743-1); 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 A05743-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05743-1) |
| Primary antibody | Rabbit anti-MEF2B, 2-5 μg/ml (datasheet A05743-1) |
| Primary incubation | Overnight at 4 °C (datasheet A05743-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A05743-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MEF2B-positive staining in germinal center cells of lymph node (HPA tissue IHC: High). HPA tissue profile: Selective nuclear expression in germinal center lymphocytes of tonsil and lymph node. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A05743-1). The published 1:200 dilution applies to the antibody used in that study (PMC11143196).
Section 2
What Is the Expected MEF2B Staining Pattern?
MEF2B is a nuclear transcription factor with no transmembrane segment (UniProt Q02080: localisation, topology). In paraffin-section IHC, expect selective, high nuclear staining in germinal center cells of tonsil and lymph node (HPA: tissue IHC). HPA rates the tissue pattern Supported because staining agrees with RNA expression, while cautioning that the antibody targets proteins from more than one gene (HPA: tissue IHC reliability).
| Strong, selective nuclear staining in tonsil or lymph-node germinal center cells. | This matches the reported high germinal center signal and nuclear pattern (HPA: tissue IHC). Score the cells and compartment together; a brown area alone does not establish the expected result (standard IHC interpretation). |
| Predominantly cytoplasmic or membrane-like staining in the expected germinal center cells. | This does not match the tissue IHC nuclear pattern or UniProt nuclear localisation (HPA: tissue IHC; UniProt Q02080: localisation). Check whether counterstain, pigment or detection background is being mistaken for specific signal (standard IHC practice). |
| Strong staining in cells reported as unstained, such as adipocytes or bone-marrow hematopoietic cells. | HPA reports MEF2B as not detected in those cell types (HPA: tissue IHC). Consider antibody cross-reactivity, consistent with HPA's multi-gene caution, or endogenous detection activity before calling the signal MEF2B (HPA: tissue IHC caution; standard IHC practice). |
| Diffuse colour over many cell types, extracellular areas or the whole section. | A diffuse field obscures the selective nuclear pattern reported for germinal centers (HPA: tissue IHC). Review blocking, washes and detection controls for general background; the appearance alone cannot identify its cause (standard IHC practice). |
| No convincing nuclear signal in a tonsil or lymph-node germinal center. | These are the supplied high-staining reference sites (HPA: tissue IHC). First confirm that the germinal center is present, then check antibody and detection controls; an absent signal alone cannot distinguish a technical failure from a sample difference (standard IHC practice). |
💡Expected MEF2B appearanceCall a positive result when germinal center cells show selective, high nuclear IHC staining (HPA: tissue IHC); broad cytoplasmic or unrelated-cell staining is suspect against that reference pattern (HPA: tissue IHC; UniProt Q02080: localisation).
| Tissue and cell selection | Tonsil and lymph-node germinal center cells provide reported high signal; selected adipocytes and bone-marrow hematopoietic cells are not detected (HPA: tissue IHC). Interpret each cell population separately. |
| Compartment | UniProt places MEF2B in the nucleus, and tissue IHC reports selective nuclear staining (UniProt Q02080: localisation; HPA: tissue IHC). The absence of a transmembrane segment offers no basis for an expected membrane pattern (UniProt Q02080: topology). |
| Antibody evidence | The listed antibody HPA004734 has Supported IHC status; HPA also warns that its tissue evidence targets proteins from more than one gene (HPA: antibody validation; HPA: tissue IHC reliability). Treat unexpected staining as needing independent support. |
| Isoforms and processing | UniProt lists 2 isoforms and a single 1–365 chain, with no signal peptide or propeptide (UniProt Q02080: isoforms, processing). These facts alone do not predict a different IHC compartment or tissue pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Germinal centers look negative while the rest of the slide develops colour. | The supplied positive reference is high nuclear staining in these cells, so this result is unresolved (HPA: tissue IHC). | Verify germinal center identity, inspect a concurrent positive control, and review antibody dilution and detection performance (standard IHC practice). |
| Many unrelated cells show nuclear signal. | This exceeds the selective tissue pattern; HPA cautions that the antibody targets proteins from more than one gene (HPA: tissue IHC). | Compare cell types with HPA's reported positives and negatives, then seek an independent antibody or orthogonal check before assigning identity (HPA: tissue IHC; standard IHC practice). |
| Staining is mainly cytoplasmic or outlines cell borders in paraffin IHC. | That differs from the reported nuclear tissue IHC pattern and UniProt localisation (HPA: tissue IHC; UniProt Q02080: localisation). | Check nuclear counterstain alignment and detection controls; record the discordance rather than scoring that colour as the expected IHC positive (standard IHC practice). |
| Background makes nuclear scoring difficult. | Diffuse detection colour can conceal selective nuclear staining (standard IHC practice; HPA: tissue IHC pattern). | Review blocking, wash steps, detection exposure and a negative detection control, then score only clearly localised nuclear signal (standard IHC practice). |
| Colour persists where primary antibody was omitted. | A primary-independent signal indicates background from the detection workflow, including possible endogenous activity (standard IHC practice). | Address the detection control first, then reassess whether germinal center nuclei retain selective staining (standard IHC practice; HPA: tissue IHC pattern). |
| Q: Why does an IF/ICC image show cytosol or cell junctions? | A: HPA approves nucleoplasm and cytosol as main IF/ICC locations and cell junctions as an additional location, with a multi-gene antibody caution (HPA: subcellular ICC-IF). | Interpret that IF/ICC observation in its own assay context; use the selective nuclear germinal center pattern to judge tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing MEF2B staining
A05743-1 · IHCIHC analysis of MEF2B using anti-MEF2B antibody (A05743-1). MEF2B was detected in a paraffin-embedded section of human hodgkin lymphoma tiss
A05743-1 · IHCIHC analysis of MEF2B using anti-MEF2B antibody (A05743-1). MEF2B was detected in a paraffin-embedded section of human tonsil tissue. Heat m
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000213999-MEF2B/tissue/cerebral+cortex#imid_1565296
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000213999-MEF2B/tissue/colon#imid_1565222
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000213999-MEF2B/tissue/liver#imid_1565207
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000213999-MEF2B/tissue/kidney#imid_1565224
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004734; green = MEF2B, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004734; green = MEF2B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MEF2B IHC & IF
🧪Run lymph node first and score germinal center cells for nuclear staining (HPA: High in lymph-node germinal center cells; UniProt Q02080: Nucleus). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the lymph-node slide, cells outside germinal centers that lack nuclear signal can serve as internal background references, without assuming every such cell is MEF2B-negative (HPA: High in germinal center cells).
Positive control tissue: Lymph node (Germinal center cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MEF2B in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and species- and clonality-matched rabbit IgG isotype controls; use MEF2B knockout material as a biological negative where available (selected IHC caption: rabbit primary antibody). Quench endogenous peroxidase and check for endogenous biotin signal in the lymph-node section when using the caption’s biotin-based DAB detection (selected IHC caption: biotinylated secondary, SABC and DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A05743-1 tissue-IHC caption does not state its fixative. Heat retrieval in EDTA at pH 8.0 was used for the paraffin-section example, but its necessity has not been established; the caption used 2 μg/ml primary antibody overnight at 4°C (selected IHC caption). The supplied evidence does not establish that frozen sections or IF are easier; for lymph-node IHC, evaluate peroxidase and biotin background with the technical controls (selected IHC caption: biotin-based DAB detection).
HPA tissue IHC evidence for MEF2B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). 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 |
|---|---|---|---|---|
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MEF2B IHC Tips
Troubleshoot MEF2B staining in paraffin-section chromogenic IHC using the catalog antibody’s tissue example and the reported cellular expression patterns.
What retrieval should I try first when nuclear MEF2B staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A05743-1). The matched tissue example used this retrieval before overnight incubation at 4°C with 2 μg/ml catalog antibody (caption A05743-1). Keep section thickness, heating, cooling, and detection conditions consistent while comparing a weak case with a positive control containing germinal center lymphocytes (HPA: high in lymph node and tonsil germinal center cells; standard IHC practice). If staining remains weak, vary retrieval duration on adjacent sections and inspect morphology for heat damage (standard IHC practice). Score nuclear signal separately from diffuse cytoplasmic color, because MEF2B is annotated as nuclear (UniProt Q02080: nucleus).
Could fixation explain absent MEF2B staining in my paraffin sections?
Target-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (caption A05743-1). Record the fixative, time in fixative, processing schedule, and section age for each case before comparing staining (standard IHC practice). Run sections with different documented fixation histories alongside the same positive control, using EDTA at pH 8.0 and 2 μg/ml catalog antibody as the matched starting conditions (datasheet A05743-1; caption A05743-1). Compare nuclear signal and preserved morphology together, since fixation and processing can affect antigen access and tissue appearance (standard IHC practice). A negative result alone cannot establish fixation sensitivity or MEF2B absence (standard IHC practice).
Should cytoplasmic or junctional staining count as MEF2B positive in IHC?
Prioritize staining in intact nuclei when scoring paraffin-section IHC, because MEF2B is annotated as nuclear (UniProt Q02080: nucleus). An HPA tissue-IHC profile describes selective nuclear staining in germinal center lymphocytes, with high staining in lymph node and tonsil germinal center cells (HPA: tissue IHC). HPA also reports nucleoplasmic, cytosolic, and junctional signals in its subcellular dataset, but cautions that those antibodies target proteins from multiple genes (HPA: subcellular). Therefore, record cytoplasmic or junctional color separately and check whether it survives control comparisons before attributing it to MEF2B (HPA: subcellular caution; standard IHC practice). Exclude folds, damaged edges, and necrotic areas from compartment assessment (standard IHC practice).
Can this antibody distinguish MEF2B isoforms or altered epitopes in sections?
Do not assign isoform identity from a positive nuclear stain: the record lists 2 MEF2B isoforms, while the selected tissue-IHC caption supplies no isoform-specific epitope (UniProt Q02080: isoforms 1 and 2; caption A05743-1). MEF2B has a MADS-box at residues 3–57, but its position alone does not locate this antibody’s binding site (UniProt Q02080: domain; caption A05743-1). Request the immunogen or epitope information before interpreting staining differences as isoform-specific, and document what that information actually covers (standard IHC practice). Compare sections under matched EDTA pH 8.0 retrieval and detection conditions to reduce technical differences (datasheet A05743-1; standard IHC practice). Corroborate an isoform claim with an independent, isoform-discriminating method (standard IHC practice).
How should I adapt this IHC result for multiplex MEF2B immunofluorescence?
Treat multiplex IF as a separate optimization: the selected antibody example documents chromogenic IHC on a paraffin section, not IF validation (caption A05743-1). Pair MEF2B with a validated germinal center lymphocyte marker and compare signal in marker-positive cells with nearby marker-negative cells (HPA: germinal center lymphocyte staining; standard IF practice). Choose fluorophores and filter sets after checking the section’s autofluorescence, reserving a brighter, spectrally separated channel for weak MEF2B signal (standard IF practice). After retrieval, optimize gentle permeabilisation for access to the nuclear epitope; MEF2B has no transmembrane segment and is annotated as nuclear (UniProt Q02080: topology and localisation; standard IF practice). Include single-color and no-primary controls before calling colocalisation (standard IF practice).
How can I reduce widespread brown staining without losing nuclear MEF2B signal?
Check a no-primary section and inspect whether brown color tracks tissue edges, folds, vessels, or damaged regions rather than nuclei (standard IHC practice; UniProt Q02080: nucleus). The selected example used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, a biotinylated secondary, and DAB development (caption A05743-1). Adjust blocking, washing, primary concentration, and chromogen development one variable at a time while retaining a positive control (standard IHC practice; HPA: high in germinal center cells). Include a peroxidase block for DAB detection and consider endogenous biotin when using the caption’s biotin-based detection scheme (standard IHC practice; caption A05743-1). Treat diffuse color without cell-specific nuclear contrast as background pending control results (standard IHC practice).
How should I quantify MEF2B across sections with different germinal center content? ⚠ ANSWER MARKED FOR VERIFICATION
Define germinal center regions and count intact, interpretable nuclei within them before scoring, because HPA reports high MEF2B staining in germinal center cells (HPA: lymph node and tonsil tissue IHC; standard IHC practice). Report the percentage of positive nuclei and, when intensity is reproducible, an H-score using intensity categories 0–3 for a range of 0–300 (standard IHC scoring practice). Normalize positive-cell counts to the number of eligible germinal center cell nuclei, or report positive nuclei per mm² of annotated region (standard IHC practice). Apply one positivity threshold and counterstain assessment across batches, with a shared control section (standard IHC practice). Exclude necrosis, folds, and section edges before calculating denominators (standard IHC practice).
What would make a brown MEF2B signal convincing rather than artefactual?
A convincing IHC pattern is discrete nuclear staining in the expected germinal center lymphocytes, consistent with the nuclear annotation and the HPA tissue profile (UniProt Q02080: nucleus; HPA: germinal center cells). Compare lymph node or tonsil germinal centers, reported as high, with cell populations reported as undetected, while interpreting differences in tissue context (HPA: tissue IHC). Be cautious with predominantly cytoplasmic or junctional color: HPA reports those subcellular signals but warns that its antibodies can target proteins from multiple genes (HPA: subcellular caution). Edge staining, necrotic areas, and color reproduced in a no-primary control favor artefact; endogenous peroxidase can also contribute to DAB color (standard IHC practice). Resolve ambiguous cases with an independently validated specificity control before assigning biological meaning (standard IHC practice).
Boster reagents
Best MEF2B / Myocyte-specific enhancer factor 2B IHC Antibodies
A05743-1 has IHC images from human paraffin-embedded Hodgkin lymphoma and tonsil sections (A05743-1 image captions). The catalog lists human, mouse, and rat reactivity (catalog: A05743-1).
Real IHC data
Anti-MEF2B Antibody ®
Cat # A05743-1
A05743-1 will render with its own IHC figure from a human paraffin-embedded Hodgkin lymphoma section (A05743-1 image caption). A second caption documents human tonsil IHC, while the catalog lists IHC use and human, mouse, and rat reactivity (A05743-1 tonsil image caption; catalog: A05743-1).
Which to pick: Choose A05743-1 for paraffin-section IHC; its captions document human Hodgkin lymphoma and tonsil sections, but do not report the fixative (A05743-1 image captions). No SKU in this payload is listed for IF/ICC (catalog: A05743-1 applications). For mouse or rat IHC, A05743-1 is the listed option, although its supplied IHC images show human tissue only (catalog: A05743-1 reactivity and applications; A05743-1 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q02080 (MEF2B_HUMAN, Myocyte-specific enhancer factor 2B).
- Human Protein Atlas. MEF2B tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MEF2B subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the cell junctions. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. MEF2B antibody validation summary (1 antibodies).
- Utility of MEF2B immunohistochemistry in distinguishing follicular lymphoma and marginal zone lymphoma in diagnostically challenging cases. Journal of hematopathology 2026 — PMC13323876.
- Spatial phenotyping of nodular lymphocyte predominant Hodgkin lymphoma and T-cell/histiocyte-rich large B-cell lymphoma. Blood cancer journal 2024 — PMC11143196.
- MEF2B Instructs Germinal Center Development and Acts as an Oncogene in B Cell Lymphomagenesis. Cancer cell 2018 — PMC6223119.
- Cx58 is associated with the metastasis of non-small cell lung cancer via MEF2B/Cx58 axis. Acta biochimica et biophysica Sinica 2025 — PMC12368521.
- PubMed PMID:1516833 — UniProt-cited evidence.
- PubMed PMID:1748287 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MEF2B (Myocyte-specific enhancer factor 2B). Expected localisation Nuclear in germinal center lymphocytes (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05743-1). Validated IHC antibody: A05743-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lymph node (Germinal center cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human hodgkin lymphoma tissue; fixative not specified (datasheet A05743-1); 2. Fixation: Image fixative and duration unreported (datasheet A05743-1); 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 A05743-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05743-1); 8. Primary antibody: Rabbit anti-MEF2B, 2-5 μg/ml (datasheet A05743-1); 9. Primary incubation: Overnight at 4 °C (datasheet A05743-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A05743-1); Caveat: Antibody may detect protein from more than one gene (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences are unspecified (UniProt); Regulation: Lymphoid tissue enriched; intensity may vary by cell population (HPA tissue IHC). Evidence: UniProt Q02080, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet A05743-1, PMC11143196. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MEF2B/immunohistochemistry-ihc-guide.