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- Table of Contents
TPM1 / Tropomyosin alpha-1 chain · Western blot design guide
Design a Western Blot for TPM1
Real validated TPM1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TPM1 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
TPM1 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 | ~32.7 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 10 isoform(s) |
Section 1
Real Curated TPM1 Western Blot Protocols
The M01591 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human heart lysate (catalog M01591) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M01591; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| 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 TPM1 Western Blot Band Size?
TPM1 is predicted at 32.7 kDa; isoforms and homodimer formation could affect migration, but no empirical band or feature-specific shift is established.
| Band near 32.7 kDa | Consistent with TPM1 at its predicted mass; confirm identity with antibody controls. |
| Higher band near twice the monomer mass | Could reflect a TPM1 homodimer if it survives sample denaturation. |
| Several discrete bands | Could reflect TPM1 splice isoforms if they differ enough in migration to resolve. |
| Band below 32.7 kDa | Could reflect a smaller TPM1 splice isoform; identity needs confirmation. |
💡Expected TPM1 appearanceTPM1 has a predicted mass of 32.7 kDa, but no empirical band size is supplied; use antibody specificity and TPM1 depletion controls to identify bands, since isoform migration and dimer persistence are unestablished.
| UniProt predicted mass | Places the reference monomer near 32.7 kDa. |
| TPM1 homodimer | Could produce a higher band near twice the monomer mass if it survives denaturation; no inter-chain disulfide is listed. |
| Splice isoforms 1, 2, 3, 4, and 5 | May differ in size, but their relative masses and band separation are not supplied. |
| Splice isoforms 6, 7, 8, 9, and 10 | May differ in size, but their relative masses and band separation are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TPM1 associates with cytoskeletal F-actin and may be poorly recovered during extraction. | Check the insoluble fraction and verify antibody performance with a positive lysate. |
| Band higher than expected | A TPM1 homodimer may persist through sample preparation. | Compare denaturing conditions and confirm the band with TPM1 depletion. |
| Band lower than expected | A smaller splice isoform is possible, but its mass is not supplied. | Check isoform recognition and confirm the band with TPM1 depletion. |
| Multiple bands | TPM1 has ten annotated splice isoforms, though their migration is not established. | Check antibody isoform coverage and compare bands after TPM1 depletion. |
| Weak or no signal | Cytoskeleton-associated TPM1 may be poorly recovered. | Inspect the insoluble fraction and include a positive lysate control. |
| Fragments below expected size | The supplied features do not establish a cleavage product. | Check sample integrity and test whether the fragments disappear after TPM1 depletion. |
Experiment evidence
Western blot images showing TPM1 band
M01591 · TPM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TPM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TPM1 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (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: Tissue lysates are suitable for cytoskeletal TPM1, but verify that adipose tissue gives no detectable band.
HPA tissue expression evidence for TPM1
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 |
|---|---|---|---|---|
| Appendix | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Colon | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | High | Protein (IHC) | HPA → |
| Rectum | enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | 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 → |
| Bone marrow | hematopoietic 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 TPM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TPM1, answered from its protein features.
What should I consider if extra TPM1 bands appear?
Band shift · Compare the bands with the 10 annotated isoforms and the antibody epitope. TPM1 is also annotated as a homodimer and as a heterodimer with TPM2, but those features alone do not identify an extra Western blot band. No observed band sizes are supplied.
Could different TPM1 isoforms produce different bands?
Isoforms · UniProt lists 10 isoforms with alternative sequences, including changes at canonical positions 1–80, 189–212 and 258–284. Check which isoforms your sample and antibody can detect before assigning multiple bands. The features alone do not establish their apparent sizes.
Check its epitope against the isoforms you intend to measure. Several isoforms replace parts of the canonical N terminus, residues 189–212 or C terminus; an antibody targeting a replaced segment may detect only some isoforms.
Use an antibody whose epitope is present in the isoforms you want to compare, and define whether you are measuring one isoform or combined TPM1 signal. UniProt lists 10 isoforms with sequence differences, so a single band may not represent every isoform.
Check its epitope against the isoforms you intend to measure. Several isoforms replace parts of the canonical N terminus, residues 189–212 or C terminus; an antibody targeting a replaced segment may detect only some isoforms.
Use an antibody whose epitope is present in the isoforms you want to compare, and define whether you are measuring one isoform or combined TPM1 signal. UniProt lists 10 isoforms with sequence differences, so a single band may not represent every isoform.
Which TPM1 modification sites matter when interpreting bands?
PTM · Canonical UniProt coordinates list N-acetylmethionine at 1; phosphoserine at 45, 174, 186, 206, 252, 271 and 283; and phosphotyrosine at 261. Serine 283 is annotated as phosphorylated by DAPK1. Confirm how your antibody or source numbers isoform residues; these sites alone do not establish a visible band shift.
TPM1 has annotated phosphorylation sites, including canonical Ser283 by DAPK1, but their presence does not prove that phosphorylation caused a shifted band. Check the detected isoform and antibody epitope before assigning the band.
TPM1 has annotated phosphorylation sites, including canonical Ser283 by DAPK1, but their presence does not prove that phosphorylation caused a shifted band. Check the detected isoform and antibody epitope before assigning the band.
Does this guide establish induction of TPM1?
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 TPM1?
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 M01591 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 TPM1 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Should TPM1 run at its predicted 32.7 kDa?
Interpretation · 32.7 kDa is the predicted mass, but no observed band size is supplied. The listed isoforms and modifications do not establish an apparent mass or explain any difference from 32.7 kDa.
Boster reagents
TPM1 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-Tropomyosin 1 Rabbit Monoclonal Antibody
Cat # M01591
The catalog reports one anti-TPM1 antibody, M01591, a rabbit monoclonal with listed human, mouse, and rat reactivity. Its Western blot image shows TPM1 expression in human heart lysate; the supplied evidence does not show mouse or rat blot results.
Which to pick: M01591 is the only listed option and has a Western blot image using human heart lysate. For mouse or rat samples, reactivity is listed, but no blot image for those samples is supplied.
Source: BosterBio TPM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P09493.
- Human Protein Atlas. TPM1 tissue expression.
- PMC4532463 — target-verified WB comparison
- PMC12167420 — target-verified WB comparison
- PMC7836198 — target-verified WB comparison