MAFA Western Blot Experimental Design Guide

Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before running
Expected band 37 kDa observed
Observed band 37 kDa observed
Gel 12-15%
Positive control No defensible HPA tissue; use knockout/knockdown
Negative control Use knockout/knockdown or orthogonal negative
Important caveats
Reasons your observed band may differ from the expected size.
Calculated mass37 kDa
LocalizationNucleus
Processing / PTMRecord-dependent
ReactivityHuman / Mouse / Rat
Section 1

Real Curated MAFA Western Blot Protocols

Start with the molecular-weight rule, then compare verified publication-derived conditions.

Recommended Western blot protocol parameters
Sample / lysateKnown-positive human lysate
Gel %12-15%
Load20-30 µg total protein per lane
TransferSemi-dry, standard transfer
Membrane0.45 µm PVDF
Blocking5% non-fat milk or 5% BSA in TBST
PrimaryA05251-1 at datasheet starting dilution
Primary incubationOvernight at 4 °C with gentle agitation
SecondarySpecies-matched HRP conjugate at validated dilution
Wash3 × 5 min in TBST
DetectionChemiluminescent substrate
ExposureBracket exposures to avoid saturation
Section 2

What Is the Expected MAFA Western Blot Band Size?

Use the product-observed 37 kDa band as the primary planning value and retain the UniProt calculated mass as context.

What am I looking at on my blot?
37 kDaMatches the authoritative product WB observation.
37 kDa calculatedUse as UniProt context, not as a replacement observed band.
Unexpected additional signalDo not assign identity without orthogonal positive/negative controls.
💡Expected MAFA appearancePlan around 37 kDa and keep the calculated mass as supporting context.
How each factor affects band size
Catalog-observed band37 kDa; use this as the primary experimental expectation.
Calculated mass37 kDa from UniProt Q8NHW3; retain as context.
Gel selection12-15%; shared with the recommended protocol and poster.
Specificity checkCompare the lead HPA positive and negative controls with A05251-1.
Why is my band missing or off?
SituationLikely causeNext action
37 kDaMatches the authoritative product WB observation.Confirm with orthogonal controls and the linked product record.
Additional bandMay reflect processing, modification, or non-specific signal.Run a dilution series and compare positive/negative controls.
Weak signalTarget abundance or transfer may be limiting.Verify transfer, increase positive-control abundance, and bracket exposure.

Sample controls for MAFA Western blot

🧪No defensible HPA positive control was available; do not invent one.
HPA protein score determines control status; other expression data is supporting context only.

HPA tissue expression evidence for MAFA

Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart on each tissue page — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue Cell type Level Evidence Source

Undetected expression · recommended negative controls

The Human Protein Atlas exposes no structured IHC tissue profile for MAFA, so tissue-level positive/negative controls cannot be listed here. Use a documented MAFA-positive lysate (recombinant MAFA or a known-expressing line) as the positive control and a MAFA knockdown/knockout lysate as the negative control; confirm against the antibody datasheet.
Tissue Cell type Level Evidence Source
Section 3

Advanced MAFA Western Blot Tips

Deeper troubleshooting and optimisation questions for MAFA, answered from its protein features.

Which band should guide the blot?
Use 37 kDa, the observation attached to the authoritative A05251-1 WB record.
How should calculated mass be interpreted?
Treat the UniProt calculated mass as context; it does not replace the catalog-observed 37 kDa expectation.
Which positive control should I start with?
No defensible positive tissue was supplied; do not invent one.
Which negative control is defensible?
Use a target knockdown/knockout control when no HPA Not detected tissue is available.
Which gel should I use?
Use 12-15% consistently across the quick facts, protocol table, and poster.
What transfer method to use for MAFA Western blot?
Use the transfer method in the recommended protocol and verify transfer before blocking.
How should A05251-1 be started?
Start at the linked datasheet condition and run a three-point primary-antibody dilution test.
Which publication-derived protocols can I compare?
No verified publication protocol was supplied; use only the deterministic recommended protocol.
Boster reagents

MAFA Western Blot Reagents

Human/Mouse/Rat-reactive MAFA Western blot reagents with authoritative product imagery.

Real WB dataWestern blot validation image for MAFA using A05251-1; observed band 37 kDa
Anti-MAFA Picoband® Antibody
Cat # A05251-1
Real WB dataWestern blot validation image for MAFA using RP1066; observed band 37 kDa
Anti-Transcription factor MafA MAFA Antibody Picoband®
Cat # RP1066

Only image-backed, WB-validated Human/Mouse/Rat recommendations from the prepared catalog evidence are shown.

Source: prepared picoband-wb product evidence; each card retains its own SKU, URL, observed band, and authoritative WB image.