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- Table of Contents
THRA / Thyroid hormone receptor alpha · Western blot design guide
Design a Western Blot for THRA
Source-linked THRA Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-THRA 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
THRA 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 | ~54.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bronchus (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 4 isoform(s) |
Section 1
Source-Linked THRA Western Blot Protocol Options
The M01775-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Jurkat cell lysate (catalog M01775-1) |
| 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 | M01775-1; 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 THRA Western Blot Band Size?
THRA is predicted at 54.8 kDa; isoforms and homodimerization could affect bands, but their effects on migration are not demonstrated here.
| Band near 54.8 kDa | Consistent with the predicted THRA mass; confirm identity with controls |
| Several bands at different positions | Could reflect Alpha-1, Alpha-2, Alpha-3, or Alpha-4; distinct migration is unconfirmed |
| One band despite four annotated isoforms | The isoforms may not resolve or may not all be detected |
| Band near twice the predicted mass | Could reflect retained THRA homodimer; confirm its identity |
💡Expected THRA appearanceTHRA has a predicted mass of 54.8 kDa, but no empirical band size is supplied; confirm a band near that size with ordinary identity controls, since isoform migration and dimer retention are unestablished.
| Predicted THRA mass | Provides a 54.8 kDa reference for a THRA band |
| Alpha-1 and Alpha-2 splice isoforms | May differ in apparent size; their relative sizes are not supplied |
| Alpha-3 and Alpha-4 splice isoforms | May differ in apparent size; their relative sizes are not supplied |
| THRA homodimer formation | Could produce a band near twice the monomer mass if the dimer survives electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear THRA may be poorly recovered or diluted in whole-cell lysate | Check nuclear extraction and a suitable positive lysate |
| Band higher than expected | A THRA homodimer could persist during electrophoresis | Compare denaturing conditions and verify THRA identity |
| Band lower than expected | An alternatively spliced THRA isoform may differ in size | Compare isoform-aware controls and verify the band |
| Multiple bands | Four THRA splice isoforms are annotated, but their migration is unknown | Use isoform-aware controls to identify each band |
| Weak or no signal | Nuclear THRA may be underrepresented in the sample | Check nuclear enrichment and antibody performance with a positive control |
Experiment evidence
Western blot images showing THRA band
M01775-1 · THRA
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for THRA Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for THRA in Western blot, you can use bronchus tissue lysate.
Positive control: Bronchus (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for THRA
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 |
|---|---|---|---|---|
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
| Prostate | glandular cells | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced THRA Western Blot Tips
Deeper troubleshooting and optimisation questions for THRA, answered from its protein features.
How should THRA 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.
Which THRA isoforms could produce different bands?
Isoforms · UniProt lists Alpha-1, Alpha-2, Alpha-3, and Alpha-4. Relative to the supplied 490-residue sequence, Alpha-1 replaces residues 371–490, while Alpha-3 lacks 371–409 and Alpha-4 lacks 371–412. These differences could affect band positions; confirm identities with an isoform-specific reagent.
Choose an epitope outside the listed variable region beginning at residue 371, and verify its sequence in each isoform. An antibody against the original 371–490 region may miss Alpha-1 or the deleted portions of Alpha-3 and Alpha-4. Coordinates here follow the supplied UniProt sequence.
Decide whether the measurement is total THRA or a particular isoform. For total THRA, verify that the antibody recognizes all four listed isoforms before combining bands. For isoform-specific quantitation, use a reagent validated against the relevant alternative sequence and quantify its band separately.
Choose an epitope outside the listed variable region beginning at residue 371, and verify its sequence in each isoform. An antibody against the original 371–490 region may miss Alpha-1 or the deleted portions of Alpha-3 and Alpha-4. Coordinates here follow the supplied UniProt sequence.
Decide whether the measurement is total THRA or a particular isoform. For total THRA, verify that the antibody recognizes all four listed isoforms before combining bands. For isoform-specific quantitation, use a reagent validated against the relevant alternative sequence and quantify its band separately.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of THRA?
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 THRA?
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 M01775-1 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 THRA 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 THRA appear at exactly 54.8 kDa?
Interpretation · 54.8 kDa is the predicted mass; no observed Western blot band is supplied. Use it as a reference, but do not assign a band by mass alone. The listed alternative sequences may affect isoform sizes, though these features do not establish apparent migration.
THRA binds DNA as a homodimer or as a heterodimer with RXRB. That interaction alone does not establish that a dimer survives Western blot preparation. Verify a higher band independently before assigning it to a THRA complex.
UniProt places THRA in the nucleus. Compare nuclear enrichment when assessing a candidate band, alongside antibody specificity and the listed isoforms. Localization can support an assignment but cannot identify a band by itself.
THRA binds DNA as a homodimer or as a heterodimer with RXRB. That interaction alone does not establish that a dimer survives Western blot preparation. Verify a higher band independently before assigning it to a THRA complex.
UniProt places THRA in the nucleus. Compare nuclear enrichment when assessing a candidate band, alongside antibody specificity and the listed isoforms. Localization can support an assignment but cannot identify a band by itself.
Boster reagents
THRA 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-THRA Rabbit Monoclonal Antibody
Cat # M01775-1
The catalog reports one anti-THRA antibody for Western blot: rabbit monoclonal M01775-1, with reported human reactivity and a WB image showing THRA expression in Jurkat cell lysate. The supplied evidence covers this example only.
Which to pick: M01775-1 is the only listed option. Its reported human reactivity and Jurkat cell lysate WB image make it the directly documented choice for that context; performance in other samples is not established here.
Source: BosterBio THRA gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P10827.
- Human Protein Atlas. THRA tissue expression.
- PMC12442676 — target-verified WB comparison
- PMC7907286 — target-verified WB comparison
- PMC8677787 — target-verified WB comparison