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- Table of Contents
MTA1 / Metastasis-associated protein MTA1 · Western blot design guide
Design a Western Blot for MTA1
Real validated MTA1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MTA1 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
MTA1 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 | ~80.8 kDa | |
| Observed band | ~80 kDa | |
| Gel | 5–20% (catalog PA1483) | |
| Positive control | Cerebellum (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated MTA1 Western Blot Protocols
The PA1483 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human HEK293, human HELA (catalog PA1483) |
| Gel % | 5–20% (catalog PA1483) |
| Load | 50ug; reducing conditions (catalog PA1483) |
| Transfer | a Nitrocellulose membrane at 150mA for 50-90 minutes (catalog PA1483) |
| Membrane | Nitrocellulose membrane (catalog PA1483) |
| Blocking | 5% Non-fat Milk/ TBS for 1.5 hour at RT (catalog PA1483) |
| Primary antibody | PA1483 · 0.5 μg/mL (catalog PA1483) |
| Primary incubation | overnight at 4°C (catalog PA1483) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PA1483) |
| Secondary incubation | 1.5 hour at RT (catalog PA1483) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1483) |
| Detection | ECL (catalog PA1483) |
Section 2
What Is the Expected MTA1 Western Blot Band Size?
UniProt predicts 80.8 kDa and antibody QC observes ~80 kDa for MTA1; the cause of the small difference is not established.
| Band near 80 kDa | Matches the reported MTA1 band in reducing whole-cell lysates. |
| Additional band at a different mass | Could reflect a splice isoform; confirm its identity. |
| Faint whole-cell band with stronger nuclear signal | Consistent with MTA1's nuclear location. |
| Doublet that collapses after phosphatase treatment | Would support a contribution from annotated phosphorylation sites. |
💡Expected MTA1 appearanceUniProt predicts 80.8 kDa, and antibody QC reports a ~80 kDa band in reducing whole-cell lysates; confirm unexpected bands with MTA1 depletion or an independent antibody.
| UniProt predicted mass | 80.8 kDa for the supplied sequence, close to the observed ~80 kDa band. |
| Long splice isoform | May differ in size from other isoforms; its mass is not supplied. |
| Short splice isoform | May differ in size from other isoforms; its mass is not supplied. |
| Isoform 3 | May differ in size from other isoforms; its mass is not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear MTA1 may be insufficiently represented in the sample. | Check a nuclear fraction and include a positive lysate control. |
| Band higher than expected | An alternate isoform or modified form is possible; its migration is unverified. | Check MTA1 depletion and compare with an independent antibody. |
| Band lower than expected | An alternate isoform or protein degradation is possible. | Check MTA1 depletion and prepare fresh lysate with protease inhibitors. |
| Multiple bands | Splice isoforms or phosphorylated forms are possible. | Use MTA1 depletion to identify specific bands; compare phosphatase-treated samples. |
| Weak or no signal | Nuclear MTA1 may be diluted in whole-cell lysate. | Check nuclear enrichment and run the antibody QC positive control. |
Experiment evidence
Western blot images showing MTA1 band
PA1483 · MTA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MTA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MTA1 in Western blot, you can use cerebellum tissue, which HPA rates as highly positive.
Positive control: Cerebellum (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: MTA1 is nuclear, so nuclear extracts may improve detection.
HPA tissue expression evidence for MTA1
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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Placenta | hofbauer cells | High | Protein (IHC) | HPA → |
| Testis | Leydig cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | Medium | 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 → |
| Cervix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MTA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MTA1, answered from its protein features.
How should MTA1 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 MTA1 isoforms produce additional bands?
Isoforms · Three isoforms are listed: Long, Short, and 3. The Short isoform replaces residues 398–430 and lacks residues 431–715, so it may appear below the full-length band. Confirm that the antibody recognizes a sequence retained in the isoform before assigning a band.
In isoform 3, residues 64–81 are replaced by one arginine. Its sequence therefore differs from the full-length sequence, but the supplied features do not establish a distinct visible band. Check whether the antibody’s target sequence includes residues 64–81.
In isoform 3, residues 64–81 are replaced by one arginine. Its sequence therefore differs from the full-length sequence, but the supplied features do not establish a distinct visible band. Check whether the antibody’s target sequence includes residues 64–81.
Which phosphorylation sites matter when interpreting MTA1 bands?
PTM · UniProt lists phosphoserines at 386, 446, 449, 522, 576, and 639, and phosphothreonines at 564 and 578. These are UniProt coordinates; antibody or paper numbering may differ. Site annotation does not establish that phosphorylation causes a detectable band shift.
Yes. UniProt lists alternate N6-acetyllysine at position 626. This is a UniProt coordinate, and the site is within residues 431–715, which are missing from the Short isoform. Its presence does not establish a visible band shift.
Yes. UniProt lists alternate N6-acetyllysine at position 626. This is a UniProt coordinate, and the site is within residues 431–715, which are missing from the Short isoform. Its presence does not establish a visible band shift.
Does this guide establish induction of MTA1?
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 MTA1?
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 PA1483 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 can MTA1 bands be quantified consistently?
Quantitation · MTA1 is listed in the nucleus and as a component of the NuRD complex. Compare equivalent sample fractions and quantify the same defined band across lanes; record whether the measurement includes the band near 80 kDa alone or additional bands assigned to isoforms.
How should the observed MTA1 band compare with predicted mass?
Interpretation · MTA1 has a predicted mass of 80.8 kDa, consistent with the reported band near 80 kDa. The listed modifications alone do not establish a visible shift or explain any difference between calculated and apparent mass.
First compare them with the band near 80 kDa and the listed isoform sequences. The Short isoform lacks residues 431–715, while isoform 3 changes residues 64–81. Check antibody target coverage before assigning either. Listed phosphorylation and acetylation sites alone cannot identify an unexpected band.
First compare them with the band near 80 kDa and the listed isoform sequences. The Short isoform lacks residues 431–715, while isoform 3 changes residues 64–81. Check antibody target coverage before assigning either. Listed phosphorylation and acetylation sites alone cannot identify an unexpected band.
Boster reagents
MTA1 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-MTA1 Antibody Picoband®
Cat # PA1483
The catalog reports one anti-MTA1 antibody, PA1483, for Western blotting. Its image shows an approximately 80 kDa band in human cell lines and rat and mouse brain lysates. No publication or independent validation evidence was supplied.
Which to pick: PA1483 is the only listed option. It is listed as reactive with human, mouse and rat, and its Western blot image includes samples from all three species. Choose it when those documented sample contexts fit your experiment.
Source: BosterBio MTA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13330.
- Human Protein Atlas. MTA1 tissue expression.
- PMC2972736 — target-verified WB comparison
- PMC6379974 — target-verified WB comparison
- PMC12704216 — target-verified WB comparison