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- Table of Contents
MTF1 / Metal regulatory transcription factor 1 · Western blot design guide
Design a Western Blot for MTF1
Source-linked MTF1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MTF1 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
MTF1 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 | ~81 kDa | |
| Observed band | ~70 kDa | |
| Gel | 10% (catalog A04733-2) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MTF1 Western Blot Protocol Options
The A04733-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human A431, human THP-1, human U251, rat liver, rat heart, mouse liver, mouse heart (catalog A04733-2) |
| Gel % | 10% (catalog A04733-2) |
| Load | 30 ug; reducing conditions (catalog A04733-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04733-2) |
| Membrane | nitrocellulose membrane (catalog A04733-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04733-2) |
| Primary antibody | A04733-2 · 1:1000 (catalog A04733-2) |
| Primary incubation | overnight at 4°C (catalog A04733-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04733-2) |
| Secondary incubation | 1.5 hour at RT (catalog A04733-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04733-2) |
| Detection | ECL (catalog A04733-2) |
Section 2
What Is the Expected MTF1 Western Blot Band Size?
MTF1 is predicted at 81 kDa, but antibody QC reports a band near 70 kDa; the cause of the difference is not established.
| Band near 70 kDa | Empirical MTF1 band reported in whole-cell lysates; confirm identity with controls |
| Band near 81 kDa | Compatible with the predicted MTF1 mass; confirm identity with controls |
| Weak band in a nuclear fraction | Compatible with cytoplasmic localization before metal-induced nuclear translocation |
| Stronger band in a nuclear fraction after metal exposure | Compatible with metal-induced nuclear translocation |
💡Expected MTF1 appearanceMTF1 has a predicted mass of 81 kDa, while antibody QC reports a band near 70 kDa in whole-cell lysates; the difference is unexplained, so verify band identity with appropriate controls.
| Predicted MTF1 mass | 81 kDa is the sequence-based reference; the reported band is near 70 kDa |
| N-acetylglycine at residue 2 | Present, but no apparent size effect is established |
| Phosphoserine at residue 5 | Present, but no apparent size effect is established |
| Phosphoserine at residue 305 | Present, but no apparent size effect is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MTF1 signal may be below detection; absence is not explained by secretion | Check loading and antibody performance with a positive-control lysate |
| Band lower than expected | The reported band is near 70 kDa versus the predicted 81 kDa; its migration difference is unexplained | Compare with the antibody QC pattern and verify identity using MTF1 depletion |
| Band higher than expected | The supplied features do not establish a higher-migrating MTF1 species | Verify identity using MTF1 depletion and check molecular-weight calibration |
| Multiple bands | Additional bands cannot be assigned to distinct isoforms from the supplied features | Identify the MTF1-dependent band using MTF1 depletion |
| Weak or no signal | MTF1 can occupy nuclear and cytoplasmic compartments, with metal-induced nuclear translocation | Check both fractions and compare metal-treated with untreated samples |
Experiment evidence
Western blot images showing MTF1 band
A04733-2 · MTF1
A04733-3 · MTF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MTF1 Western blot
🧪For positive controls for MTF1 in Western blot, you can use no HPA-supported positive sample because tissue expression data were not supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: HPA provides no tissue data, so validate any positive sample experimentally and use knockdown or KO for a negative control.
HPA tissue expression evidence for MTF1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced MTF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MTF1, answered from its protein features.
How should MTF1 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 MTF1 isoforms explain multiple bands?
Isoforms · Only one isoform is listed, with no alternative sequence. The supplied features therefore do not support assigning multiple bands to different MTF1 isoforms.
Which MTF1 modifications matter when interpreting bands?
PTM · UniProt lists N-acetylglycine at position 2 and phosphoserine at positions 5 and 305, using its sequence numbering. Their presence alone does not establish a visible band shift.
No glycosylation sites are listed. The supplied features do not support attributing an unexpected band or the 70 versus 81 kDa difference to MTF1 glycosylation.
No glycosylation sites are listed. The supplied features do not support attributing an unexpected band or the 70 versus 81 kDa difference to MTF1 glycosylation.
Does this guide establish induction of MTF1?
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.
How should transfer be checked for MTF1?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04733-2 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 nuclear MTF1 be quantified after metal exposure?
Quantitation · Compare like fractions across conditions and normalize each fraction consistently. Metal-induced nuclear translocation means a change in nuclear signal alone does not establish a change in total MTF1 abundance.
Why might MTF1 appear near 70 kDa instead of 81 kDa?
Interpretation · The supplied observed band is about 70 kDa, while the predicted mass is 81 kDa. These features do not explain the difference or establish the band’s identity. Confirm the band with an independent MTF1-specific check.
The supplied location note says metals induce MTF1 translocation to the nucleus. If comparing metal exposure conditions, keep exposure and fraction collection consistent and examine nuclear and cytoplasmic samples when testing localization.
Do not assign them to isoforms or phosphorylation solely by position: one isoform and phosphoserines at UniProt positions 5 and 305 are listed, but neither establishes a distinct band. Check whether each band tracks with an independent MTF1-specific measurement.
The supplied location note says metals induce MTF1 translocation to the nucleus. If comparing metal exposure conditions, keep exposure and fraction collection consistent and examine nuclear and cytoplasmic samples when testing localization.
Do not assign them to isoforms or phosphorylation solely by position: one isoform and phosphoserines at UniProt positions 5 and 305 are listed, but neither establishes a distinct band. Check whether each band tracks with an independent MTF1-specific measurement.
Boster reagents
MTF1 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-MTF1 Antibody
Cat # A04733-2
Real WB data
Anti-MTF1 Antibody Picoband®
Cat # A04733-3
The catalog reports two anti-MTF1 antibodies with Western blot images from human cell lysates and rat and mouse tissues. Both captions report bands below the expected 81 kDa: approximately 70 kDa for A04733-2 and 65 kDa for A04733-3. Independent validation is not supplied.
Which to pick: Both list human, mouse, and rat reactivity and have WB images. Choose A04733-2 for a closer match to the reported A431, THP-1, or U251 cell samples; choose A04733-3 for 293T or brain tissue samples. Check the reported band size against your sample.
Source: BosterBio MTF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14872.
- Human Protein Atlas. MTF1 tissue expression.
- PMC10307746 — target-verified WB comparison
- PMC12160861 — target-verified WB comparison
- PMC6299381 — target-verified WB comparison