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MAF / Transcription factor Maf · IHC design guide
Design Immunohistochemistry for MAF
Plan paraffin-section MAF IHC around the nuclear pattern reported in most tissues (HPA tissue IHC). Compare cell-specific staining with appropriate controls, bearing in mind the reported low agreement between antibody staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MAF 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 most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear signal across most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Fallopian tube+1 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 | Antibody staining has low RNA concordance (HPA tissue IHC) | |
| Regulation | Staining intensity varies by cell type (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended MAF IHC & IF Protocols
The catalog antibody’s IHC-P protocol is paired with one published human tissue microarray protocol for nuclear MAF staining (PMC8301809; UniProt O75444: nucleus).
| Sample | Paraffin-embedded human kidney tissue; fixative not specified (datasheet M00654) |
| Fixation | Image fixative and duration unreported (datasheet M00654); 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 monoclonal (clone DHI-13) anti-MAF, 1:50 (datasheet M00654) |
| 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 | MAF-positive staining in glial cells of caudate (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page antigen-retrieval setting; UniProt O75444: nucleus).
Section 2
What Is the Expected MAF Staining Pattern?
MAF is a nuclear transcription factor with no transmembrane segment (UniProt O75444: nucleus; topology). In paraffin tissue sections, expect predominantly nuclear staining across many tissues, including glial cells, kidney tubular cells, and selected glandular cells (HPA: nuclear expression in most tissues; listed high staining). HPA rates its tissue IHC profile Approved but reports low consistency between antibody staining and RNA expression; interpret intensity and tissue comparisons with that caveat (HPA: reliability description).
| Distinct nuclear staining in glial cells of caudate or cerebral cortex, or kidney tubular cells. | This matches the expected compartment and documented high-staining cell populations (UniProt O75444: nucleus; HPA: High in caudate/cortex glial cells and kidney tubules). Judge the stained cells, rather than treating the whole tissue as uniformly positive (HPA: cell-level tissue entries). |
| Predominantly cytoplasmic staining, with little or no nuclear signal, in a tissue expected to stain. | This conflicts with the predominant tissue IHC pattern (HPA: nuclear expression in most tissues) and UniProt localization (UniProt O75444: nucleus). Consider nonspecific staining or detection artefact; do not classify cytoplasmic staining alone as a convincing MAF-positive result. |
| Strong glandular staining in fallopian tube or seminal vesicle. | These glandular cells are listed as Not detected (HPA: fallopian tube and seminal vesicle glandular cells). Check whether staining follows a tissue-associated background pattern or persists in a no-primary control (general IHC practice). HPA's Approved profile has low RNA–staining consistency, so this is a comparison, not proof of cross-reactivity (HPA: reliability description). |
| Diffuse chromogen across nuclei, cytoplasm, and extracellular spaces, obscuring cell boundaries. | An indiscriminate deposit cannot establish the expected nuclear pattern (HPA: nuclear expression in most tissues; UniProt O75444: nucleus). Excess detection background, incomplete blocking, or insufficient washing are possible general IHC causes (general IHC practice); use controls and a cleaner stain before scoring. |
| No nuclear signal in a high-staining reference tissue. | This disagrees with the listed cell-specific observations, such as kidney tubules or caudate glia (HPA: High in those cells). First verify that the relevant cells are present and the assay worked with controls (general IHC practice). A blank section alone does not establish absent MAF expression. |
💡Expected MAF appearanceCall positive when clearly stained nuclei occur in the relevant cells, potentially at high intensity in HPA-listed populations; diffuse background or isolated cytoplasmic staining is unconvincing (UniProt O75444: nucleus; HPA: nuclear profile and listed High cells).
| Cell-level tissue choice | Use a listed high-staining population, such as glial cells in caudate or tubular cells in kidney, to judge whether the nuclear pattern is visible (HPA: High in these cells). Fallopian tube and seminal vesicle glandular cells provide documented Not detected comparisons, with the HPA reliability caveat applied (HPA: tissue entries; low RNA–staining consistency). |
| Strength of tissue evidence | The HPA tissue IHC profile is Approved and reports nuclear expression in most tissues, while noting low consistency between staining and RNA expression (HPA: reliability and profile). Both listed antibodies have Approved IHC status; neither is listed here as Enhanced (HPA: antibody validation entries). Avoid treating a single intensity difference as a definitive expression ranking. |
| Isoforms and antibody coverage | UniProt lists two MAF isoforms (UniProt O75444: isoforms 1 and 2). The supplied antibody entries give no epitope or isoform coverage (HPA: antibody entries). A tissue pattern cannot identify which isoform was detected; consult antibody-specific documentation before making an isoform claim. |
| Protein architecture | MAF is a 373-residue protein with a bZIP region at residues 288–351, one listed chain, no signal peptide, and no transmembrane segment (UniProt O75444: sequence, domain, processing, topology). Those features support a nuclear interpretation, but do not specify an antibody epitope or predict antigen-retrieval performance. |
| Detection background | Endogenous enzyme activity and nonspecific detection can create chromogen unrelated to the primary antibody (general IHC practice). Interpret unexpected widespread staining with a no-primary control and suitable detection controls (general IHC practice); the supplied HPA tissue profile does not identify a MAF-specific background mechanism. |
| IF/ICC Q&A: Where should signal appear? | Mainly in nucleoplasm and nuclear bodies, with additional Golgi apparatus and vesicle localization reported in ICC-IF (HPA: approved subcellular locations). This separate IF observation does not make predominantly cytoplasmic paraffin IHC staining the expected tissue result (HPA: nuclear expression in most tissues). |
| Situation | Likely cause | Next action |
|---|---|---|
| High-staining reference tissue appears blank. | The relevant cell population may be absent from the viewed field, or staining and detection may have failed (HPA: cell-specific High entries; general IHC practice). | Locate the documented cells, inspect the counterstain and assay controls, then review the IHC-validated antibody's documented procedure and repeat with a suitable positive control (general IHC practice). |
| Signal appears mainly cytoplasmic in paraffin sections. | The observed compartment disagrees with the expected nuclear tissue pattern (HPA: nuclear expression in most tissues; UniProt O75444: nucleus); background is one possible cause (general IHC practice). | Compare with a no-primary control, assess whether nuclei are discernible, and optimize detection and washing using standard IHC controls before assigning positivity (general IHC practice). |
| Every cell and extracellular area carries brown deposit. | Widespread deposit is compatible with excessive chromogen development or nonspecific detection (general IHC practice), rather than the documented cell-associated nuclear profile (HPA: tissue profile). | Check a no-primary control; review blocking, washes, detection reagents, and development time under the chosen IHC method (general IHC practice). Score only a resolved cellular pattern. |
| Fallopian tube or seminal vesicle glandular cells stain strongly. | This differs from their Not detected HPA entries (HPA: tissue IHC), although the Approved profile has low staining–RNA consistency (HPA: reliability description). | Confirm cell identity and nuclear location, compare controls, and check whether the same run reproduces a listed high-staining pattern (HPA: tissue entries; general IHC practice). Treat discordance as unresolved until verified. |
| One high-staining tissue is weaker than another. | HPA lists high staining in particular cell types but does not establish identical signal across tissues or runs (HPA: tissue entries). Section composition and assay variation can affect comparisons (general IHC practice). | Compare matched cell populations with the same staining run and scoring criteria, and record nuclear intensity separately from the fraction of positive cells (general IHC practice). |
| IF/ICC shows puncta outside nuclei. | Golgi apparatus and vesicle localization are additionally reported alongside nucleoplasm and nuclear bodies (HPA: approved ICC-IF subcellular locations). | Interpret the IF image against its nuclear and additional compartment observations (HPA: ICC-IF summary), while using the predominantly nuclear tissue profile to score paraffin IHC (HPA: tissue IHC profile). |
Experiment evidence
IHC & IF images showing MAF staining
M00654 · IHCImmunohistochemical analysis of paraffin-embedded human kidney, using c-Maf Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000178573-MAF/tissue/cerebral+cortex#imid_3667504
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000178573-MAF/tissue/colon#imid_3667422
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000178573-MAF/tissue/liver#imid_3667423
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000178573-MAF/tissue/kidney#imid_3667424
HPA · ICC-IF AF22Human Protein Atlas immunofluorescence, AF22, antibody HPA028289; green = MAF, blue = nucleus, red = microtubules.
HPA · ICC-IF AF22Human Protein Atlas immunofluorescence, AF22, antibody HPA028289; green = MAF, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MAF IHC & IF
🧪Run caudate first and look for MAF staining in glial cells (HPA: High in caudate glial cells); use fallopian tube glandular cells as the negative tissue (HPA: Not detected in fallopian tube glandular cells). On the caudate slide, compare glial nuclear staining with unstained neighboring nuclei as an internal background reference, without treating those cells as validated MAF-negative cells (UniProt O75444: nucleus; HPA: High in caudate glial cells).
Positive control tissue: Caudate (Glial cells, HPA High)
Negative control tissue: Fallopian tube (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAF in AF22, Rh30, U2OS, with annotated localisation: Nucleoplasm (approved), Nuclear bodies (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, a host-species- and Ig-class-matched isotype control for a monoclonal antibody or host-matched normal IgG for a polyclonal antibody, and a matched MAF-knockout section if available (standard IHC practice). For chromogenic caudate IHC, quench endogenous peroxidase before HRP detection and check the no-primary slide for background (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the M00654 paraffin-section caption does not state a fixative (supplied target/application evidence; selected M00654 tissue-IHC caption). MAF-specific retrieval dependence is unreported; optimize retrieval empirically for nuclear staining in paraffin sections (UniProt O75444: nucleus; standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; for caudate IF, check tissue autofluorescence with a no-primary control (supplied target/application evidence; standard IF practice).
HPA tissue IHC evidence for MAF
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced MAF IHC Tips
Troubleshoot c-Maf staining in paraffin section IHC by checking nuclear signal, tissue context, and controls before changing the detection workflow.
How should I retrieve c-Maf in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Allow slides to cool consistently before applying the IHC-validated antibody, and compare weak sections with a positive kidney tubule control processed in the same run (HPA: High in kidney tubule cells). If staining remains weak, test a citrate buffer at pH 6.0 on adjacent sections as a fallback, keeping antibody concentration and detection constant (standard IHC optimisation). Score nuclear signal separately from background, since c-Maf is a nuclear transcription factor (UniProt O75444: Nucleus).
Could fixation explain weak or uneven c-Maf staining?
Target-specific fixation sensitivity is unknown: the selected M00654 image identifies paraffin-embedded human kidney but does not report its fixative (M00654 tissue-IHC caption). Record each specimen’s fixative and time in fixative, then compare sections processed with the same retrieval and detection settings (standard IHC practice). For a controlled optimisation, compare routinely fixed sections with deliberately varied fixation durations, such as 6 and 24 h, while holding tissue thickness constant (standard IHC optimisation). Interpret differences as effects of the tested preparation until replicated; neither the tissue staining profile nor the protein record establishes a c-Maf-specific fixation response (HPA: tissue IHC; UniProt O75444: protein record).
Where should convincing c-Maf staining appear in tissue sections?
Prioritise a signal within nuclei: c-Maf is annotated as nuclear, and tissue IHC reports nuclear expression in most tissues (UniProt O75444: Nucleus; HPA: tissue IHC). In kidney, examine tubule cell nuclei because those cells are reported at a High staining level; the selected antibody also has a paraffin-embedded kidney image (HPA: kidney tubule cells; M00654 tissue-IHC caption). Use a hematoxylin counterstain to separate nuclear chromogen from adjacent cytoplasm and luminal deposits (standard chromogenic IHC practice). Additional Golgi and vesicle localisation comes from cell imaging, so diffuse cytoplasmic tissue staining alone should trigger a specificity check (HPA: subcellular ICC/IF).
How do unknown epitope and isoform coverage affect c-Maf IHC?
The record lists 2 isoforms, but the supplied antibody evidence does not specify its epitope or isoform coverage (UniProt O75444: isoforms; M00654 tissue-IHC caption). Request the immunogen or epitope map before treating staining differences as evidence for one isoform; compare any mapped sequence against both isoforms (standard antibody validation practice). The bZIP region spans residues 288–351, which matters only if the antibody’s mapped epitope overlaps it (UniProt O75444: bZIP domain). With no signal peptide, transmembrane segment, or annotated glycosylation sites, interpret unexpected membrane staining cautiously and check it with an independent antibody (UniProt O75444: topology and glycosylation).
How can IF help check the c-Maf pattern seen by IHC?
Use IF on a matched section to check whether c-Maf signal overlaps nuclei, since the protein is nuclear and cell imaging places it mainly in nucleoplasm and nuclear bodies (UniProt O75444: Nucleus; HPA: subcellular ICC/IF). Multiplex with DAPI and an endothelial marker such as CD31 to assess an expected cell population; endothelial expression is reported for c-Maf (UniProt O75444: tissue specificity; standard IF marker practice). Choose a far-red fluorophore, such as a 647 nm channel, and include an unstained section to assess tissue autofluorescence (standard IF practice). Because the target is intracellular and nuclear, start with mild permeabilisation, such as 0.1% Triton X-100 for 5 min, then optimise against preserved nuclear morphology (UniProt O75444: localisation; standard IF practice).
What controls distinguish c-Maf chromogen from background?
Run a no-primary control through the same secondary and chromogenic detection steps to locate reagent-dependent staining (standard IHC practice). Block endogenous peroxidase before DAB development, and compare the resulting precipitate with the no-primary section; these are general chromogenic workflow steps, not c-Maf-specific validation (standard IHC practice). Reduce excess primary antibody or detection reagent only after confirming that the positive kidney tubule nuclei remain visible (HPA: High in kidney tubule cells; standard IHC optimisation). Inspect tissue edges, folds, and damaged areas separately because uneven reagent access can create local signal unrelated to nuclear antigen distribution (standard IHC interpretation).
How should I score variable nuclear c-Maf staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define eligible cell compartments before scoring, then report the percentage of nuclei positive for c-Maf and the number of scored nuclei per section (UniProt O75444: Nucleus; standard IHC scoring). If intensity is reliable, calculate a nuclear H-score from 0–300 using percentages at intensity grades 0–3 (standard IHC scoring). For spatial comparisons, report positive nuclei per mm² and normalise to the area of viable, evaluable tissue or to all eligible nuclei in the same compartment (standard IHC quantification). Keep kidney tubule cells separate from other kidney cells because the reported High signal is assigned specifically to tubule cells (HPA: kidney tubule cells).
When is a c-Maf-positive cell credible rather than an artefact?
A credible result has discrete nuclear signal in an identifiable, intact cell and survives comparison with a no-primary control (UniProt O75444: Nucleus; standard IHC interpretation). Kidney tubule nuclei provide a documented positive context, while glandular cells in fallopian tube and seminal vesicle were reported as Not detected in the cited tissue survey (HPA: tissue IHC). Distrust staining restricted to section edges, necrotic regions, lumina, or cytoplasm without corresponding nuclei; check peroxidase-block and no-primary controls for chromogen artefact (standard IHC interpretation). Treat tissue patterns cautiously because the tissue atlas labels its antibody staining Approved while reporting low consistency with RNA expression (HPA: reliability description).
Boster reagents
Best MAF / Transcription factor Maf IHC Antibodies
The IHC antibody has an image from a human kidney paraffin section and lists human, mouse and rat reactivity (M00654 image caption; M00654 catalog reactivity). A separate antibody lists human IF/ICC use (A00654-1 catalog applications and reactivity).
Real IHC data
Anti-c-Maf Rabbit Monoclonal Antibody
Cat # M00654
M00654 is the SKU that will render; its IHC figure shows a paraffin-embedded human kidney section (M00654 image caption). Its listed application is IHC, with human, mouse and rat reactivity (M00654 catalog applications and reactivity).
Which to pick: Choose M00654 for tissue IHC: it is a rabbit monoclonal listed for IHC, and its own image caption documents a paraffin-embedded human kidney section; the fixative is unreported (M00654 catalog host, clone and applications; M00654 image caption). For IF/ICC, choose A00654-1, which lists those applications and human reactivity (A00654-1 catalog applications and reactivity). For mouse or rat IHC, M00654 is the listed cross-species option, although its own IHC figure shows human tissue (M00654 catalog reactivity and image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75444 (MAF_HUMAN, Transcription factor Maf).
- Human Protein Atlas. MAF tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MAF subcellular location (ICC-IF): Mainly localized to the nucleoplasm and nuclear bodies. In addition localized to the Golgi apparatus and vesicles..
- Human Protein Atlas. MAF antibody validation summary (2 antibodies).
- Sequential and combinatorial roles of maf family genes define proper lens development. Molecular vision 2007 — PMC2503189.
- c-MAF deletion in adult C57BL/6J mice induces cataract formation and abnormal differentiation of lens fiber cells. Experimental animals 2020 — PMC7220708.
- Characterization of c-Maf transcription factor in normal and neoplastic hematolymphoid tissue and its relevance in plasma cell neoplasia. American journal of clinical pathology 2009 — PMC4847138.
- Loss of the MAF Transcription Factor in Laryngeal Squamous Cell Carcinoma. Biomolecules 2021 — PMC8301809.
- PubMed PMID:9616139 — UniProt-cited evidence.
- PubMed PMID:15616553 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MAF (Transcription factor Maf). Expected localisation Nuclear staining in most tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: M00654 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Glial cells); negative Fallopian tube. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human kidney tissue; fixative not specified (datasheet M00654); 2. Fixation: Image fixative and duration unreported (datasheet M00654); 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 monoclonal (clone DHI-13) anti-MAF, 1:50 (datasheet M00654); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody staining has low RNA concordance (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Staining intensity varies by cell type (HPA tissue IHC). Evidence: UniProt O75444, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and nuclear bodies. In addition…, Datasheet M00654, PMC8301809. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MAF/immunohistochemistry-ihc-guide.