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- Table of Contents
TBR1 / T-box brain protein 1 · Western blot design guide
Design a Western Blot for TBR1
Real validated TBR1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TBR1 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
TBR1 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 | ~74.1 kDa | |
| Observed band | ~74 kDa | |
| Gel | 5–20% (catalog A07503-2) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) | |
| 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
Real Curated TBR1 Western Blot Protocols
The A07503-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SH-SY5Y, rat brain, rat C6 (catalog A07503-2) |
| Gel % | 5–20% (catalog A07503-2) |
| Load | 30 ug; reducing conditions (catalog A07503-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07503-2) |
| Membrane | nitrocellulose membrane (catalog A07503-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07503-2) |
| Primary antibody | A07503-2 · 0.5 μg/mL (catalog A07503-2) |
| Primary incubation | overnight at 4°C (catalog A07503-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07503-2) |
| Secondary incubation | 1.5 hour at RT (catalog A07503-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07503-2) |
| Detection | ECL (catalog A07503-2) |
Section 2
What Is the Expected TBR1 Western Blot Band Size?
TBR1 is predicted at 74.1 kDa and observed at ~74 kDa; the small difference has no established cause.
| Band at ~74 kDa | Empirical TBR1 band, close to its predicted 74.1 kDa mass; confirm identity with controls |
| Band near ~148 kDa | Could reflect the reported homodimer if it survives sample preparation |
| Multiple bands near the expected region | Could reflect isoforms 1 and 2, though distinct migration is unproven |
| Weak band in whole-cell lysate | Nuclear localization may make TBR1 easier to detect in an enriched fraction |
💡Expected TBR1 appearanceTBR1 is predicted at 74.1 kDa and observed at ~74 kDa; use appropriate positive samples and band-identity controls to verify the signal.
| Predicted TBR1 mass | 74.1 kDa, consistent with the observed ~74 kDa band |
| Homodimer | Could yield a band near twice the monomer mass if preserved |
| Splice isoform 1 | May differ in apparent size from isoform 2; its individual mass is unknown |
| Splice isoform 2 | May differ in apparent size from isoform 1; its individual mass is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TBR1 is nuclear, and the sampled lysate may contain little detectable target | Check a nuclear fraction and a positive brain tissue sample |
| Band higher than expected | The reported homodimer may persist after incomplete dissociation | Compare reducing and nonreducing preparations |
| Band lower than expected | An isoform is possible, but its migration is unknown | Confirm band identity with an independent antibody or target depletion |
| Multiple bands | Isoforms 1 and 2 are annotated, but distinct bands are unverified | Check which bands track with target depletion |
| Weak or no signal | The nuclear target may be scarce in the chosen sample | Use a positive brain tissue sample and assess nuclear enrichment |
Experiment evidence
Western blot images showing TBR1 band
A07503-2 · TBR1
M07503 · TBR1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TBR1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TBR1 in Western blot, you can use cerebral cortex tissue, which HPA reports as positive.
Positive control: Cerebral cortex (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: TBR1 is nuclear and has medium HPA signal in cerebral cortex, so nuclear enrichment may improve detection.
HPA tissue expression evidence for TBR1
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 |
|---|---|---|---|---|
| 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TBR1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TBR1, answered from its protein features.
How should TBR1 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 TBR1 isoforms produce different bands?
Isoforms · Yes. UniProt lists isoforms 1 and 2; isoform 2 lacks residues 1–287 of the canonical sequence. It could therefore migrate below the full-length band. Check whether the antibody recognizes a region retained in isoform 2 before assigning a lower band.
Which TBR1 phosphorylation sites matter when interpreting a blot?
PTM · UniProt lists phosphothreonine 408 and phosphoserines 410, 594, and 641. These are canonical-sequence coordinates; isoform 2 lacks the first 287 residues, so its numbering differs. Phosphorylation alone does not establish a visible mobility shift.
Does this guide establish induction of TBR1?
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 TBR1?
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 A07503-2 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.
What sample fraction is most relevant for TBR1 quantitation?
Quantitation · TBR1 is listed as nuclear. Use a consistent nuclear sample preparation and loading basis across lanes when comparing its signal; differences in nuclear recovery could otherwise affect the measured band intensity.
How does the observed TBR1 band compare with its predicted mass?
Interpretation · The observed band near 74 kDa agrees with the predicted 74.1 kDa for the 682-residue canonical protein. The listed features do not establish a measurable band shift.
TBR1 is reported to form a homodimer and to participate in a protein complex. A higher band may warrant checking whether sample preparation preserved protein associations, but these features alone cannot identify that band.
Consider isoform 2, which lacks canonical residues 1–287. Confirm that the antibody targets a retained region and compare the lower band with the full-length signal before assigning it to an isoform.
TBR1 is reported to form a homodimer and to participate in a protein complex. A higher band may warrant checking whether sample preparation preserved protein associations, but these features alone cannot identify that band.
Consider isoform 2, which lacks canonical residues 1–287. Confirm that the antibody targets a retained region and compare the lower band with the full-length signal before assigning it to an isoform.
Boster reagents
TBR1 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-TBR1 Antibody Picoband®
Cat # A07503-2
Real WB data
Anti-TBR1 Rabbit Monoclonal Antibody
Cat # M07503
Both listed anti-TBR1 antibodies report human, mouse, and rat reactivity and have Western blot images. A07503-2 shows a band near the expected 74 kDa in named cell and brain lysates; M07503 shows TBR1 expression in human fetal brain lysate. No publication evidence is supplied.
Which to pick: Choose A07503-2 for the documented human SH-SY5Y, rat brain or C6, and mouse brain or Neuro-2a blot conditions. Choose M07503 if human fetal brain lysate better matches your sample; its supplied caption gives fewer experimental details.
Source: BosterBio TBR1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16650.
- Human Protein Atlas. TBR1 tissue expression.
- PMC12078632 — target-verified WB comparison
- PMC6414196 — target-verified WB comparison