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- Table of Contents
MAFK / Transcription factor MafK · Western blot design guide
Design a Western Blot for MAFK
Source-linked MAFK Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MAFK WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
MAFK Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~17.5 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Placenta (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MAFK Western Blot Protocol Options
The M06288 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | lysate from Jurkat cell line, (catalog M06288) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M06288; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | goat anti-mouse IgG, 1:3000 (catalog M06288) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected MAFK Western Blot Band Size?
MAFK has a predicted monomer mass of 17.5 kDa; dimerization and Ser25 phosphorylation are annotated, but their effects on blot migration are unestablished.
| Band near 17.5 kDa | consistent with monomeric MAFK; confirm identity with antibody controls |
| Band near 35 kDa | could reflect a MAFK homodimer if it survives sample preparation |
| Higher band of variable size | could reflect a retained heterodimer with a CNC-bZIP partner |
| Weak band in whole-cell lysate | nuclear MAFK may be underrepresented |
💡Expected MAFK appearanceUniProt predicts a 17.5 kDa MAFK monomer; no empirical band size is supplied, so verify a band near that size with ordinary identity controls.
| UniProt predicted mass | 17.5 kDa for the MAFK monomer |
| MAFK homodimer formation | could yield a band near twice the monomer size if the complex survives sample preparation |
| Heterodimer formation with CNC-bZIP proteins | could yield a higher band if the complex survives sample preparation; partner mass is unspecified |
| Phosphoserine 25 | may affect migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear MAFK may be poorly represented in the preparation | check a nuclear fraction and use an antibody identity control |
| Band higher than expected | a MAFK dimer may persist during sample preparation | compare denaturing conditions and verify MAFK identity |
| Band lower than expected | possible degradation or an unrelated antibody-reactive protein | prepare fresh lysate with protease inhibitors and verify band identity |
| Multiple bands | possible retained dimers or unrelated antibody-reactive proteins | compare sample preparation conditions and confirm the MAFK band with an identity control |
| Weak or no signal | nuclear MAFK may be diluted in whole-cell lysate | enrich the nuclear fraction and check sample loading |
| Fragments below expected size | possible proteolysis during sample preparation | prepare fresh lysate with protease inhibitors and verify fragment identity |
Experiment evidence
Western blot images showing MAFK band
M06288 · MAFK
A06288 · MAFK
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MAFK Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MAFK in Western blot, you can use placenta lysate, which shows high HPA expression.
Positive control: Placenta (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: MAFK is nuclear, so nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for MAFK
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Placenta | trophoblastic cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAFK Western Blot Tips
Deeper troubleshooting and optimisation questions for MAFK, answered from its protein features.
How should MAFK band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Could MAFK isoforms explain multiple bands?
Isoforms · Only one MAFK isoform is listed, with no alternative sequence supplied. The supplied features therefore do not support assigning additional bands to isoforms.
How should phosphorylation be considered when interpreting MAFK bands?
PTM · UniProt lists phosphoserine at residue 25 of the supplied 156-residue sequence. Keep that UniProt coordinate explicit when comparing antibody or paper numbering. This site establishes a modification, but does not show whether it produces a resolvable band shift.
Does this guide establish induction of MAFK?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for MAFK Western blot?
Transfer · MAFK is predicted to be 17.5 kDa. Choose and check transfer conditions for retention and detection of a protein of that size; the supplied features do not specify a membrane or transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M06288 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should MAFK be quantified across samples?
Quantitation · MAFK is listed as nuclear. Use a consistent sample fraction and loading basis across samples, especially when comparing nuclear preparations. The supplied features provide no induction condition or expected change in abundance.
Why might MAFK migrate away from its predicted mass?
Interpretation · MAFK has a predicted mass of 17.5 kDa, but no observed band position is supplied. UniProt lists phosphoserine at position 25 and no glycosylation sites. That feature alone cannot establish a visible shift or explain a difference between apparent and predicted mass.
Compare candidate bands with the predicted 17.5 kDa mass, but do not treat that value as an observed band position. MAFK can form homodimers and heterodimers with CNC-bZIP proteins; the supplied features do not establish that either causes a particular Western blot band. Only one isoform is listed.
Compare candidate bands with the predicted 17.5 kDa mass, but do not treat that value as an observed band position. MAFK can form homodimers and heterodimers with CNC-bZIP proteins; the supplied features do not establish that either causes a particular Western blot band. Only one isoform is listed.
Boster reagents
MAFK Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-MAFK Antibody
Cat # M06288
Real WB data
Anti-Transcription factor MafK MAFK Antibody
Cat # A06288
Both listed anti-MAFK antibodies have Western blot images using human cell lysates: M06288 with Jurkat and A06288 with K562. The supplied evidence includes no mouse Western blot example or publication data.
Which to pick: For human samples, choose based on the closer tested context: M06288 was shown with Jurkat lysate, and A06288 with K562 lysate. A06288 also lists mouse reactivity, but its supplied Western blot image uses human cells.
Source: BosterBio MAFK gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O60675.
- Human Protein Atlas. MAFK tissue expression.
- PMC12948957 — target-verified WB comparison
- PMC3711329 — target-verified WB comparison
- PMC3847324 — target-verified WB comparison