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MYEF2 / Myelin expression factor 2 · IHC design guide
Design Immunohistochemistry for MYEF2
Plan MYEF2 paraffin IHC around its general nuclear staining pattern (HPA tissue IHC). Cerebral cortex glia stain strongly, while smooth muscle cells are reported as undetected; use these observations to guide control selection (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MYEF2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | General nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across multiple cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Smooth muscle |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA show medium concordance (HPA tissue IHC) | |
| Regulation | No specific expression regulator annotated (UniProt) | |
| Isoform / epitope | 4 isoforms; check antibody epitope coverage (UniProt) |
Section 1
Recommended MYEF2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by one published chromogenic IHC protocol for MYEF2 in HCC tissues (PMC9854206).
| Sample | Paraffin-embedded Mouse brain tissue; fixative not specified (datasheet A11666) |
| Fixation | Image fixative and duration unreported (datasheet A11666); 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-MYEF2, 1:50-1:200 (datasheet A11666) |
| 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 | MYEF2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min (page retrieval: nuclear antigen); citrate pH 6.0 with microwave heating is published for HCC tissues (PMC9854206).
Section 2
What Is the Expected MYEF2 Staining Pattern?
MYEF2 is a nuclear protein with no transmembrane segment (UniProt Q9P2K5 topology). In paraffin sections, expect predominantly nuclear staining in glandular, hematopoietic, respiratory epithelial and selected brain cells reported as high by HPA (HPA tissue IHC). The tissue pattern is rated Supported, with medium consistency between staining and RNA expression and external verification pending (HPA tissue IHC).
| Clear nuclear staining in adrenal glandular cells, bone marrow hematopoietic cells or cerebral cortex glial cells. | This matches the reported high staining in those cell populations and the general nuclear pattern (HPA tissue IHC). Judge the signal within the named cells; staining elsewhere in the section does not establish the same result. Compare signal with a negative control before scoring it as specific (general IHC practice). |
| Strong, predominantly cytoplasmic staining with little nuclear signal in an expected positive cell population. | This conflicts with nuclear localization (UniProt Q9P2K5; HPA tissue IHC) and warrants investigation of staining or detection artefact (general IHC practice). HPA reports additional cytosolic localization by ICC-IF, so a cytosolic component alone does not prove an artefact (HPA subcellular). |
| Strong staining of smooth muscle cells, especially when the expected positive cells stain weakly. | HPA reports MYEF2 as not detected in smooth muscle cells (HPA tissue IHC). Check for antibody cross-reactivity or endogenous detection activity before interpreting this as MYEF2; the supplied evidence does not identify which, if either, caused the signal (general IHC practice). |
| Broad color deposition across nuclei, cytoplasm and extracellular areas without clear cell boundaries. | A diffuse pattern cannot be scored confidently as the reported nuclear expression (HPA tissue IHC). Background from the staining or detection workflow is a possible explanation; use the negative control and tissue morphology to distinguish it from localized signal (general IHC practice). |
| No nuclear signal in an expected high-staining population, such as bronchial respiratory epithelial cells. | This is discordant with HPA's high result for those cells (HPA tissue IHC). First assess tissue preservation, controls and the IHC workflow (general IHC practice). Because HPA rates the tissue pattern Supported with external verification pending, one negative section cannot by itself settle MYEF2 expression (HPA tissue IHC). |
💡Expected MYEF2 appearanceCall a section positive when identifiable expected cells show predominantly nuclear signal, with strong staining plausible in HPA high populations; strong smooth muscle staining or cytoplasmic-only staining is a warning pattern requiring controls, rather than a secure positive call (HPA tissue IHC; UniProt Q9P2K5; general IHC practice).
| Expected compartment | UniProt places MYEF2 in the nucleus, and HPA describes general nuclear tissue staining (UniProt Q9P2K5; HPA tissue IHC). Score the named cell populations and their nuclear signal together; intensity without the expected location is insufficient for a confident call (general IHC practice). |
| Tissue and cell population | HPA reports high staining in several glandular, hematopoietic and brain cell populations, low staining in vaginal squamous epithelial cells and skeletal myocytes, and no detection in smooth muscle cells (HPA tissue IHC). Choose controls by the reported cell population within a tissue, not by tissue name alone. |
| Evidence strength | The supplied antibody HPA004883 has Supported IHC status; HPA describes medium agreement between tissue staining and RNA and says external verification is pending (HPA antibodies; HPA tissue IHC). Treat an unexpected compartment or cell type as a finding to check, not as established MYEF2 biology. |
| Isoforms and epitope coverage | UniProt lists 4 MYEF2 isoforms (UniProt Q9P2K5). The supplied record does not map the IHC antibody epitope, so it cannot establish which isoforms contribute to staining. Do not explain a tissue difference by isoform recognition without separate evidence. |
| IF/ICC: where should signal appear? | HPA reports approved nucleoplasmic localization plus additional approved cytosolic localization in ICC-IF, with images from A-431, U-251MG, U2OS and KOLF2.1J (HPA subcellular). Its separate ICC antibody status is Approved (HPA antibodies); these observations do not define an IHC-P protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| An expected high-staining population has no visible nuclear signal. | The section or staining workflow may have failed; a biological absence is also possible. HPA's high call is Supported, with external verification pending (HPA tissue IHC). | Review morphology and the run's positive control, then check retrieval, primary-antibody incubation and detection steps against the chosen IHC-P procedure (general IHC practice). Avoid assigning a MYEF2-specific retrieval requirement from this record. |
| Signal appears only in cytoplasm in an IHC section. | The result diverges from general nuclear tissue staining (HPA tissue IHC; UniProt Q9P2K5). HPA's additional cytosolic ICC-IF location does not establish a cytoplasmic-only IHC pattern (HPA subcellular). | Inspect nuclear counterstain and cell boundaries, then compare with the positive and negative controls (general IHC practice). Record any mixed nuclear and cytoplasmic signal separately from a cytoplasmic-only result. |
| Smooth muscle cells stain strongly. | HPA lists smooth muscle cells as not detected (HPA tissue IHC). Cross-reactivity or endogenous detection activity could explain unexpected color, but this record does not distinguish them. | Compare an adjacent negative control and assess whether the color persists without primary antibody; review endogenous activity blocking where relevant to the detection chemistry (general IHC practice). |
| Weak color spreads across much of the section. | Diffuse background can obscure the nuclear pattern used for interpretation (HPA tissue IHC; general IHC practice). The payload provides no MYEF2-specific cause for background. | Check negative-control staining, blocking, washes and detection development, then assess nuclei in identifiable expected cells (general IHC practice). Avoid calling diffuse color positive solely because an HPA tissue is listed as high. |
| Low-staining cells look as dark as an expected high-staining population. | That contrast differs from HPA's low calls for vaginal squamous epithelial cells and skeletal myocytes versus its listed high populations (HPA tissue IHC). Section-to-section color intensity alone is not a validated expression measurement. | Compare the named cell populations within appropriately controlled sections and use consistent scoring criteria (general IHC practice). Keep HPA's Supported reliability and pending external verification in the interpretation (HPA tissue IHC). |
| A researcher wants to apply an IF/ICC image pattern directly to IHC-P scoring. | HPA reports nucleoplasm plus additional cytosol for ICC-IF, while its tissue IHC profile is general nuclear expression (HPA subcellular; HPA tissue IHC). The observations come from different applications. | Use nuclear staining in the reported tissue cell populations as the IHC-P reference (HPA tissue IHC). Consult the separate IF/ICC guide for that application; do not infer an IF protocol or an IHC-P cytoplasmic scoring rule from these records. |
Experiment evidence
IHC & IF images showing MYEF2 staining
A11666 · IHCImmunohistochemistry of paraffin-embedded Mouse brain using MYEF2 Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrieval with
A11666 · IHCImmunohistochemistry of paraffin-embedded Rat testis using MYEF2 Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrieval with 1
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000104177-MYEF2/tissue/cerebral+cortex#imid_1725992
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000104177-MYEF2/tissue/colon#imid_1725918
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000104177-MYEF2/tissue/liver#imid_1725911
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000104177-MYEF2/tissue/kidney#imid_1725928
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004883; green = MYEF2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA004883; green = MYEF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MYEF2 IHC & IF
🧪Run adrenal gland first: its glandular cells should show MYEF2 staining (HPA: High in adrenal gland glandular cells). Run smooth muscle as the negative tissue (HPA: Not detected in smooth muscle cells); on the adrenal slide, assess neighboring cells without nuclear signal as internal background references rather than assuming every cell is negative (UniProt Q9P2K5: Nucleus).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Smooth muscle (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MYEF2 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and matched rabbit IgG controls for the rabbit polyclonal catalog antibody (selected A11666 caption: Rabbit pAb; standard IHC practice), plus MYEF2 knockout tissue or a validated peptide-block control if available (standard IHC practice). Quench endogenous peroxidase and check adrenal pigment against the control slides before scoring chromogenic signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11666 paraffin-section caption does not state a fixative (selected A11666 caption). Retrieval dependence is unestablished; that caption specifies microwave retrieval in 10 mM PBS, pH 7.2, before mouse-brain IHC at 1:100 (selected A11666 caption). The supplied evidence does not establish that frozen sections or IF/ICC are easier; in adrenal sections, assess pigment as a possible chromogenic artefact (standard IHC practice).
HPA tissue IHC evidence for MYEF2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Pending external verification.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MYEF2 IHC Tips
Troubleshoot MYEF2 staining by assessing retrieval, nuclear localisation, antibody epitope coverage and cell type before comparing chromogenic IHC results (UniProt Q9P2K5; HPA tissue IHC).
What retrieval should I try first for weak MYEF2 nuclear staining?
Use Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval setting: nuclear antigen). If staining remains weak, compare a separate section retrieved by microwave in 10 mM PBS at pH 7.2, the method shown for catalog antibody A11666 in paraffin-embedded mouse brain (A11666 tissue-IHC caption). Keep antibody concentration and detection conditions identical across those sections so retrieval is the main variable (standard IHC practice). Score nuclear signal and tissue damage together: MYEF2 is annotated as nuclear, while torn sections or diffuse staining cannot establish improved target detection (UniProt Q9P2K5 localisation; standard IHC practice).
How can I assess whether fixation is causing weak MYEF2 staining?
MYEF2-specific fixation sensitivity is unknown from the supplied evidence; the A11666 mouse-brain caption identifies paraffin sections but does not state a fixative (A11666 tissue-IHC caption). Record the fixative and fixation interval for every specimen, then compare sections processed and retrieved together (standard IHC practice). For a controlled trial, compare appropriately fixed material against material with a documented longer fixation interval, using Tris-EDTA pH 9.0 retrieval for 20 min in both groups (page retrieval setting; standard IHC practice). Judge nuclear staining alongside morphology and a processing control; neither HPA staining patterns nor MYEF2 topology establishes how this antibody responds to fixation (HPA tissue IHC; UniProt Q9P2K5 topology; standard IHC practice).
How should I troubleshoot strong MYEF2 staining outside nuclei?
First compare the chromogenic pattern with MYEF2's annotated nuclear localisation and HPA's general nuclear tissue pattern (UniProt Q9P2K5 localisation; HPA tissue IHC). HPA subcellular imaging places MYEF2 mainly in nucleoplasm and additionally in cytosol, so limited cytoplasmic signal warrants evaluation rather than automatic exclusion (HPA subcellular). Check a nuclear counterstain and inspect intact cells at high magnification; pigment, precipitate and overlapping cells can mimic extranuclear DAB signal (standard IHC practice). MYEF2 has no transmembrane segment, so prominent membrane outlining is unexpected and should prompt comparison with a no-primary control and antibody titration (UniProt Q9P2K5 topology; standard IHC practice).
Could isoform coverage explain inconsistent MYEF2 staining between specimens?
MYEF2 has 4 annotated isoforms, but the supplied evidence does not identify the catalog antibody's epitope or establish which isoforms it detects (UniProt Q9P2K5 isoforms; A11666 tissue-IHC caption). Compare the antibody's immunogen sequence with isoform sequences before interpreting a negative nucleus as absent MYEF2 (standard IHC practice). The protein contains 3 RNA recognition motifs, and reported modifications include phosphorylation and arginine methylation; their effect on this antibody's IHC staining is unknown (UniProt Q9P2K5 domains and modified residues). Keep retrieval at Tris-EDTA pH 9.0 for 20 min while evaluating epitope coverage, and use an independently validated antibody if coverage remains uncertain (page retrieval setting; standard IHC practice).
How should an IF experiment check the nuclear IHC pattern?
Treat IF/ICC as a separate assay and pair MYEF2 with a glial-cell marker when examining the glial populations reported in caudate or cerebral cortex (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and compare an unstained section or cell preparation for autofluorescence before assigning a weak MYEF2 signal, particularly in the green channel (standard IF practice). MYEF2 lacks a transmembrane segment and is mainly nucleoplasmic, so permeabilise fixed cells sufficiently to reach the nuclear antigen, then compare a no-primary control (UniProt Q9P2K5 topology; HPA subcellular; standard IF practice). Evaluate nuclear overlap with a DNA counterstain and record acquisition settings consistently; the mouse-brain paraffin IHC caption supplies no IF fixation or dilution conditions (standard IF practice; A11666 tissue-IHC caption).
What controls help identify diffuse brown background in MYEF2 IHC?
Compare the stained section with no-primary and detection-only controls, and examine whether brown signal follows nuclei or instead coats tissue edges and damaged areas (standard IHC practice; UniProt Q9P2K5 localisation). For peroxidase-based DAB detection, include an endogenous peroxidase block and inspect a matched control for residual enzyme activity (standard chromogenic IHC practice). Titrate the catalog antibody around the caption's 1:100 mouse-brain dilution while holding Tris-EDTA pH 9.0 retrieval for 20 min constant (A11666 tissue-IHC caption; page retrieval setting; standard IHC practice). That dilution is documented for the pictured paraffin specimen; compare nuclear signal and background together before applying it to other material (A11666 tissue-IHC caption; standard IHC practice).
How should I quantify MYEF2 positivity across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define a nuclear scoring rule before viewing the groups, then report either the percentage of positive nuclei or a nuclear H-score based on staining intensity and positive-cell fraction (standard IHC quantification practice; UniProt Q9P2K5 localisation). Use the same threshold, illumination and analysis settings for every section, and exclude folds, edges and necrotic regions by a prespecified rule (standard IHC quantification practice). Normalise positive nuclei to the total count of evaluable nuclei within each annotated cell population, or report positive-cell density per mm² of viable tissue (standard IHC quantification practice). Keep glandular, hematopoietic and glial compartments separate where present because HPA reports high staining in examples of each (HPA tissue IHC; standard IHC quantification practice).
When is a MYEF2-positive IHC result convincing rather than artefactual?
A convincing result shows reproducible staining of intact nuclei with low control background, consistent with MYEF2's nuclear annotation and HPA's general nuclear tissue pattern (UniProt Q9P2K5 localisation; HPA tissue IHC; standard IHC practice). HPA reports high staining in caudate glial cells and no detection in smooth-muscle cells, useful contextual comparisons within the reported material rather than universal positive or negative rules (HPA tissue IHC). Review apparent positives at section edges, in necrosis or over endogenous peroxidase activity against matched controls before scoring them (standard chromogenic IHC practice). Interpret a negative result alongside retrieval and processing controls; HPA rates its tissue evidence as supported with medium RNA–protein consistency and pending external verification (page retrieval setting; HPA tissue IHC; standard IHC practice).
Boster reagents
Best MYEF2 / Myelin expression factor 2 IHC Antibodies
Anti-MYEF2 IHC images show paraffin-embedded mouse brain and rat testis (A11666 IHC captions). IF/ICC is listed for human, mouse and rat, but no IF image is supplied (A11666 catalog).
Real IHC data
Anti-Myelin expression factor 2 MYEF2 Antibody
Cat # A11666
A11666 has IHC images of paraffin-embedded mouse brain and rat testis at 1:100 (A11666 IHC captions); IF/ICC is listed without an IF image (A11666 catalog). A11666-1 lists human paraffin-section IHC at 2–5 μg/ml and human IF/ICC at 5 μg/ml, but supplies no IHC or IF image (A11666-1 catalog).
Which to pick: For tissue IHC, pick A11666 when a paraffin-section example matters: its rabbit polyclonal antibody has mouse brain and rat testis IHC images, with microwave retrieval in 10 mM PBS, pH 7.2 (A11666 catalog and IHC captions). For human IF/ICC, either SKU is listed, while A11666-1 specifies 5 μg/ml for human samples (A11666 and A11666-1 catalogs). For cross-species work, A11666 lists human, mouse and rat reactivity and IHC/IF/ICC applications; its IHC captions document paraffin sections but do not report the fixative (A11666 catalog and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9P2K5 (MYEF2_HUMAN, Myelin expression factor 2).
- Human Protein Atlas. MYEF2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. MYEF2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. MYEF2 antibody validation summary (1 antibodies).
- Expression and function of myelin expression factor 2 in hepatocellular carcinoma. BMC gastroenterology 2023 — PMC9854206.
- MYEF2: an immune infiltration-related prognostic factor in IDH-wild-type glioblastoma. Aging 2023 — PMC10457068.
- PARP1-mediated PARylation activity is essential for oligodendroglial differentiation and CNS myelination. Cell reports 2021 — PMC9586836.
- PubMed PMID:10718198 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16572171 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MYEF2 (Myelin expression factor 2). Expected localisation General nuclear staining in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A11666 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Smooth muscle. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Mouse brain tissue; fixative not specified (datasheet A11666); 2. Fixation: Image fixative and duration unreported (datasheet A11666); 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-MYEF2, 1:50-1:200 (datasheet A11666); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA show medium concordance (HPA tissue IHC); Isoform / epitope: 4 isoforms; check antibody epitope coverage (UniProt); Regulation: No specific expression regulator annotated (UniProt). Evidence: UniProt Q9P2K5, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A11666, PMC9854206. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MYEF2/immunohistochemistry-ihc-guide.