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- Table of Contents
TPT1 / Translationally-controlled tumor protein · Western blot design guide
Design a Western Blot for TPT1
Source-linked TPT1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TPT1 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
TPT1 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 | ~19.6 kDa | |
| Gel | 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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked TPT1 Western Blot Protocol Options
The M03414 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HeLa cell lysate (catalog M03414) |
| 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 | M03414; 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 TPT1 Western Blot Band Size?
TPT1 has a predicted 19.6 kDa monomer; isoforms and homodimerization could affect patterns, but no empirical migration or distinct isoform bands are established.
| Single band near 19.6 kDa | Consistent with the predicted TPT1 monomer; confirm identity with antibody controls |
| Upper band near twice the monomer size | Could reflect a homodimer if it survives sample preparation |
| Multiple bands near the monomer region | Could reflect isoforms 1 and 2, but distinct migration is unestablished |
| Upper band disappears after stronger denaturation | Supports dissociation of the annotated homodimer |
💡Expected TPT1 appearanceUniProt predicts a 19.6 kDa TPT1 monomer; no empirical band size is supplied, and antibody specificity and sample preparation should be checked when assigning a band.
| Predicted TPT1 mass | Sets a 19.6 kDa reference for the monomer, not a measured migration |
| Homodimer | Could appear near twice the monomer size if it survives sample preparation |
| Isoform 1 | May migrate differently from isoform 2; its size is unspecified |
| Isoform 2 | May migrate differently from isoform 1; its size is unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic TPT1 may be poorly recovered or below detection | Check lysate preparation, loading, and a positive control |
| Band higher than expected | The annotated homodimer may persist during sample preparation | Compare stronger denaturing conditions and confirm band identity |
| Band lower than expected | An alternate isoform may migrate differently; its size is unknown | Confirm identity with an independent antibody or TPT1 depletion |
| Multiple bands | Isoforms 1 and 2 or persistent homodimer may contribute | Compare denaturing conditions and confirm bands with TPT1 depletion |
| Weak or no signal | Insufficient recovery or antibody recognition of cytoplasmic TPT1 | Verify loading and antibody performance with a positive control |
Experiment evidence
Western blot images showing TPT1 band
M03414 · TPT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TPT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TPT1 in Western blot, you can use appendix tissue lysate.
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: TPT1 is cytoplasmic, but HPA rates the listed positive tissues only Medium, so expect a moderate signal.
HPA tissue expression evidence for TPT1
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 | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | ciliated cells (cilia axoneme) | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal 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 → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | hepatocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TPT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TPT1, answered from its protein features.
Where should the main TPT1 band appear?
Band shift · Canonical TPT1 has a predicted mass of 19.6 kDa. Use this as a reference, since no observed band position is supplied. The listed phosphorylation sites alone do not establish a visible shift or explain any difference between apparent and predicted mass.
Could TPT1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 1–34 of the canonical sequence, so it is expected to be smaller. If two bands appear, compare their sizes with this deletion; band position alone cannot identify either isoform.
Which TPT1 phosphorylation sites matter when interpreting bands?
PTM · The supplied UniProt features list phosphoserine at positions 46, 53, and 64. PLK1 is specified for positions 46 and 64. These are canonical UniProt coordinates; paper or antibody numbering may differ. Their presence does not establish that phosphorylation causes a visible band shift.
PLK1 is listed for canonical TPT1 phosphoserines 46 and 64. A shifted band cannot be predicted from those annotations alone. If testing a PLK1-related change, compare matched samples and verify the proposed phosphorylation effect separately.
PLK1 is listed for canonical TPT1 phosphoserines 46 and 64. A shifted band cannot be predicted from those annotations alone. If testing a PLK1-related change, compare matched samples and verify the proposed phosphorylation effect separately.
Does this guide establish induction of TPT1?
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 TPT1?
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 M03414 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 multiple TPT1 bands be quantified?
Quantitation · Report which band or bands were measured. Isoform 2 lacks canonical residues 1–34, and three phosphorylation sites are listed, but the features do not establish which forms an antibody detects or whether phosphorylation separates them on a blot. Quantify bands separately until their identities are supported.
Could an unexpected high TPT1 band be a dimer?
Interpretation · TPT1 is annotated as a homodimer by similarity, but that annotation does not identify a high band on a Western blot. Treat dimerization as a possibility to investigate, while checking band identity experimentally.
Boster reagents
TPT1 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-TCTP Rabbit Monoclonal Antibody
Cat # M03414
M03414 is a rabbit monoclonal anti-TCTP antibody listed as reactive with human, mouse, and rat. Its supplied Western blot image shows a HeLa cell lysate; no mouse or rat blot is shown.
Which to pick: M03414 is the only listed antibody. Its stated reactivity covers human, mouse, and rat, while the supplied Western blot example uses HeLa cell lysate. Check suitability for other sample types.
Source: BosterBio TPT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P13693.
- Human Protein Atlas. TPT1 tissue expression.
- PMC7851619 — target-verified WB comparison
- PMC6500995 — target-verified WB comparison
- PMC5604218 — target-verified WB comparison