This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
BTRC / F-box/WD repeat-containing protein 1A · Western blot design guide
Design a Western Blot for BTRC
Source-linked BTRC Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BTRC 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
BTRC 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 | ~68.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Spleen (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked BTRC Western Blot Protocol Options
The A01747 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of U-251MG cells, (catalog A01747) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A01747) |
| Primary antibody | A01747 · 1:1000 (catalog A01747) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A01747) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A01747) |
Section 2
What Is the Expected BTRC Western Blot Band Size?
BTRC has a predicted monomer mass of 68.9 kDa; its isoforms and homodimer could affect migration, but no empirical band size is supplied.
| Band near 68.9 kDa | consistent with the predicted BTRC monomer |
| Higher band near twice the monomer size | could reflect a BTRC homodimer if the complex is preserved |
| More than one band | could reflect isoforms 1 and 2, though distinct migration is unconfirmed |
| One band despite two annotated isoforms | isoforms may comigrate or only one may be present |
💡Expected BTRC appearanceBTRC has a predicted monomer mass of 68.9 kDa, but no empirical band size is supplied; confirm any candidate band with BTRC depletion or an independent antibody.
| Predicted BTRC monomer mass | places the reference band near 68.9 kDa |
| BTRC homodimer | may produce a band near twice the monomer size if preserved during electrophoresis |
| Isoform 1 | may migrate differently from isoform 2; relative size is unknown |
| Isoform 2 | may migrate differently from isoform 1; relative size is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BTRC may be distributed between cytoplasm and nucleus | check both fractions and verify sample loading |
| Band higher than expected | a BTRC homodimer may be preserved | compare complex-preserving and denaturing conditions |
| Band lower than expected | isoform-dependent migration is possible but unconfirmed | test band identity by BTRC depletion or an independent antibody |
| Multiple bands | isoforms 1 and 2 or preserved homodimer may contribute | compare denaturing conditions and confirm bands by BTRC depletion |
| Weak or no signal | cytoplasmic and nuclear distribution may reduce signal in one fraction | compare fractions and check loading controls |
Experiment evidence
Western blot images showing BTRC band
A01747 · BTRC
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BTRC Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BTRC in Western blot, you can use appendix tissue lysate.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Spleen (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA reports medium detection in appendix and no detection in spleen, so these provide a feasible positive and negative pair.
HPA tissue expression evidence for BTRC
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 → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Endometrium | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced BTRC Western Blot Tips
Deeper troubleshooting and optimisation questions for BTRC, answered from its protein features.
How should BTRC 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 BTRC isoforms produce different bands?
Isoforms · Two isoforms are listed. Isoform 2 lacks residues 17–52 of the canonical sequence, so it has 36 fewer residues. This supports a possible size difference, but does not establish that both isoforms are expressed or resolved on a blot.
Check whether the antibody recognizes residues 17–52, which are missing in isoform 2. An antibody against that segment would not be expected to recognize isoform 2. Verify the antibody epitope and compare band positions cautiously.
Check whether the antibody recognizes residues 17–52, which are missing in isoform 2. An antibody against that segment would not be expected to recognize isoform 2. Verify the antibody epitope and compare band positions cautiously.
Do BTRC modifications explain an unexpected band?
PTM · No modified residues or glycosylation sites are supplied. The ubiquitin-like conjugation pathway keyword does not identify a modification on BTRC or establish a visible shift. Investigate band identity before assigning an unexpected band to a modification.
Does this guide establish induction of BTRC?
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 BTRC?
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 A01747 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 BTRC 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 BTRC run at exactly 68.9 kDa?
Interpretation · 68.9 kDa is the supplied predicted mass; no observed band position is available. Use it as a reference, but do not infer a band shift from the listed features alone.
BTRC self-associates and forms a homodimer within the SCF(BTRC) complex. That feature alone does not establish that a dimer will persist during Western blot preparation or produce a higher band.
BTRC is listed in both cytoplasm and nucleus. When comparing fractions, keep sample loading and normalization consistent and interpret changes alongside fraction purity; a change in one fraction may reflect distribution rather than total abundance.
The two listed isoforms offer one possibility: isoform 2 lacks canonical residues 17–52. Self-association is also reported, but does not establish a persistent blot band. Compare apparent sizes and antibody epitope coverage before assigning identities; no observed band positions are supplied.
BTRC self-associates and forms a homodimer within the SCF(BTRC) complex. That feature alone does not establish that a dimer will persist during Western blot preparation or produce a higher band.
BTRC is listed in both cytoplasm and nucleus. When comparing fractions, keep sample loading and normalization consistent and interpret changes alongside fraction purity; a change in one fraction may reflect distribution rather than total abundance.
The two listed isoforms offer one possibility: isoform 2 lacks canonical residues 17–52. Self-association is also reported, but does not establish a persistent blot band. Compare apparent sizes and antibody epitope coverage before assigning identities; no observed band positions are supplied.
Boster reagents
BTRC 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-BTRC Antibody
Cat # A01747
The catalog reports A01747, an anti-BTRC antibody with reported Human, Mouse, and Rat reactivity. Its WB image shows U-251MG cell extracts tested at 1:1000 with 25 µg per lane; the supplied evidence shows this sample context only.
Which to pick: A01747 is the only listed option and has a WB image from U-251MG extracts. Check the reported species reactivity against your sample; the supplied image does not document WB performance in other specimens.
Source: BosterBio BTRC gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9Y297.
- Human Protein Atlas. BTRC tissue expression.
- PMC12540224 — target-verified WB comparison
- PMC11850103 — target-verified WB comparison
- PMC4747187 — target-verified WB comparison