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SMARCB1 / SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 · Western blot design guide
Design a Western Blot for SMARCB1
Real validated SMARCB1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SMARCB1 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
SMARCB1 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 | ~44.1 kDa | |
| Observed band | ~44 kDa | |
| Gel | 5–20% (catalog A00500-2) | |
| Positive control | Adrenal gland (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 | Phosphorylated + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated SMARCB1 Western Blot Protocols
The A00500-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human 293T, human K562 (catalog A00500-2) |
| Gel % | 5–20% (catalog A00500-2) |
| Load | 30 ug (catalog A00500-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00500-2) |
| Membrane | nitrocellulose membrane (catalog A00500-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00500-2) |
| Primary antibody | A00500-2 · 0.5 μg/mL (catalog A00500-2) |
| Primary incubation | overnight at 4°C (catalog A00500-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00500-2) |
| Secondary incubation | 1.5 hour at RT (catalog A00500-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00500-2) |
| Detection | ECL (catalog A00500-2) |
Section 2
What Is the Expected SMARCB1 Western Blot Band Size?
SMARCB1 is predicted at 44.1 kDa and observed at ~44 kDa; isoforms A and B are annotated, but distinct migration is unestablished.
| Band at ~44 kDa | Matches the observed SMARCB1 band and its 44.1 kDa predicted mass |
| Single sharp band near 44 kDa | Consistent with SMARCB1; isoforms need not resolve separately |
| Bands at different positions | Could reflect isoforms A and B, but their migration is unestablished |
| Little or no band in a cytoplasmic fraction | Consistent with SMARCB1's nuclear location |
💡Expected SMARCB1 appearanceSMARCB1 has a predicted mass of 44.1 kDa and an observed band at ~44 kDa; confirm band identity with appropriate controls because isoform migration is not established.
| Predicted SMARCB1 mass | 44.1 kDa predicts a band near the observed ~44 kDa |
| Splice isoform A | Its individual mass and migration are not supplied |
| Splice isoform B | Its individual mass and migration are not supplied |
| Alternative splicing into A and B | May change band positions, but separate bands are not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SMARCB1 may be underrepresented in the sampled nuclear material | Check nuclear recovery and a validated positive lysate |
| Band higher than expected | The identity or migration of this band is unestablished | Compare with the ~44 kDa band and confirm specificity with SMARCB1 depletion |
| Band lower than expected | Isoform migration or band identity is unestablished | Confirm specificity with SMARCB1 depletion and an independent antibody |
| Multiple bands | Isoforms A and B are annotated, but separate bands are unconfirmed | Test band identity with SMARCB1 depletion or isoform-specific controls |
| Weak or no signal | Low SMARCB1 abundance or inadequate nuclear protein recovery | Check loading, nuclear recovery, and a validated positive lysate |
| Fragments below expected size | Fragment identity is unestablished | Check sample integrity and confirm specificity with SMARCB1 depletion |
Experiment evidence
Western blot images showing SMARCB1 band
A00500-2 · SMARCB1
M00500 · SMARCB1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SMARCB1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SMARCB1 in Western blot, you can use adrenal gland tissue, which has high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No clean negative tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for SMARCB1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Testis | Leydig cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced SMARCB1 Western Blot Tips
Deeper troubleshooting and optimisation questions for SMARCB1, answered from its protein features.
How should SMARCB1 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 SMARCB1 isoforms produce different bands?
Isoforms · UniProt lists isoforms A and B; isoform B lacks residues 69–77. Check whether your antibody recognizes both isoforms when interpreting closely spaced bands. The sequence difference alone does not establish that two bands will resolve.
How should phosphorylation affect SMARCB1 band interpretation?
PTM · UniProt lists phosphoserine at canonical-sequence position 129. Because isoform B lacks residues 69–77, residue numbering may differ by isoform or antibody source. The listed phosphorylation does not, by itself, demonstrate a visible mobility shift.
Does this guide establish induction of SMARCB1?
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.
What transfer method to use for SMARCB1 Western blot?
Transfer · SMARCB1 has a predicted mass of 44.1 kDa. Choose a transfer protocol validated around 44 kDa and check transfer at that size with a marker or total-protein stain. The supplied features do not establish a preference between transfer methods.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00500-2 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.
How should SMARCB1 bands be quantified?
Quantitation · SMARCB1 is nuclear, so keep sample preparation and nuclear protein loading consistent across comparisons. Quantify signal within the assay’s linear range. If isoforms A and B resolve and the antibody detects both, state whether you measured each band or their combined signal.
Why is the observed SMARCB1 band near 44 kDa?
Interpretation · The observed band near 44 kDa agrees with the predicted mass of 44.1 kDa. The listed features do not establish a modification-related band shift.
Consider isoform B’s missing residues 69–77 when assessing a nearby lower band, but verify band identity before assigning it to an isoform. UniProt lists phosphoserine at canonical position 129, yet that feature alone cannot explain an additional band. The listed features provide no signal peptide, propeptide, or glycosylation site to support those explanations.
Consider isoform B’s missing residues 69–77 when assessing a nearby lower band, but verify band identity before assigning it to an isoform. UniProt lists phosphoserine at canonical position 129, yet that feature alone cannot explain an additional band. The listed features provide no signal peptide, propeptide, or glycosylation site to support those explanations.
Boster reagents
SMARCB1 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-SNF5/SMARCB1 Antibody Picoband®
Cat # A00500-2
Real WB data
Anti-SNF5 SMARCB1 Rabbit Monoclonal Antibody
Cat # M00500
Two the supplier anti-SMARCB1 antibodies have WB images. A00500-2 shows an approximately 44 kDa band in human, monkey COS-7, and rat C6 lysates; M00500 shows blots of HeLa and K562 lysates. No publication evidence is supplied.
Which to pick: Both list human, mouse, and rat reactivity. Choose A00500-2 for a WB image with reported band size and detailed conditions, including rat C6 lysate; choose M00500 for a monoclonal antibody with a HeLa and K562 WB image. Neither caption shows mouse lysate.
Source: BosterBio SMARCB1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q12824.
- Human Protein Atlas. SMARCB1 tissue expression.
- PMC12144088 — target-verified WB comparison
- PMC10602591 — target-verified WB comparison
- PMC7312701 — target-verified WB comparison