This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
PBRM1 / Protein polybromo-1 · Western blot design guide
Design a Western Blot for PBRM1
Real validated PBRM1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PBRM1 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
PBRM1 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 | ~192.9 kDa | |
| Observed band | ~240 kDa | |
| Gel | 8% (catalog A01130-1) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) +4 more | |
| Negative control | Liver (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 9 isoform(s) |
Section 1
Real Curated PBRM1 Western Blot Protocols
The A01130-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human K562, human Hela, human U2OS (catalog A01130-1) |
| Gel % | 8% (catalog A01130-1) |
| Load | 30 ug; reducing conditions (catalog A01130-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01130-1) |
| Membrane | nitrocellulose membrane (catalog A01130-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01130-1) |
| Primary antibody | A01130-1 · 0.5 μg/mL (catalog A01130-1) |
| Primary incubation | overnight at 4°C (catalog A01130-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01130-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01130-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01130-1) |
| Detection | ECL (catalog A01130-1) |
Section 2
What Is the Expected PBRM1 Western Blot Band Size?
PBRM1 is predicted at 192.9 kDa, but an empirical band appears near 240 kDa; the cause of the difference is not established.
| Band near 240 kDa | Matches the empirical PBRM1 band; confirm identity with knockdown or an independent antibody. |
| Band near 193 kDa | Near the predicted sequence mass, but PBRM1 identity requires validation. |
| Several bands at different positions | PBRM1 has nine named isoforms, although distinct band positions are not established. |
| Weak band in whole-cell lysate | PBRM1 is nuclear; check recovery in a nuclear fraction. |
💡Expected PBRM1 appearancePBRM1 has a predicted mass of 192.9 kDa, while antibody QC reports a band at approximately 240 kDa; the cause of this difference is unestablished, so confirm band identity with knockdown or an independent antibody.
| Predicted PBRM1 mass | 192.9 kDa is the calculated sequence mass, while the reported band migrates near 240 kDa. |
| Splice isoforms 1, 2, and 3 | Their apparent sizes could differ; individual masses and migration positions are not supplied. |
| Splice isoforms 4, 5, and 6 | Their apparent sizes could differ; individual masses and migration positions are not supplied. |
| Splice isoforms 7, 8, and 9 | Their apparent sizes could differ; individual masses and migration positions are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear PBRM1 may be poorly recovered or insufficiently abundant in the lysate. | Check extraction and loading; compare a nuclear fraction with a positive-control lysate. |
| Band higher than expected | The reported approximately 240 kDa band exceeds the 192.9 kDa predicted mass; the cause is unestablished. | Confirm identity by PBRM1 knockdown or an independent antibody and check molecular-weight markers. |
| Band lower than expected | An isoform or protein breakdown is possible, but the band's identity is unknown. | Check sample handling and confirm PBRM1 specificity by knockdown or an independent antibody. |
| Multiple bands | Nine PBRM1 isoforms are listed, but their migration positions are unknown. | Use knockdown or an independent antibody to identify PBRM1 bands. |
| Weak or no signal | Low PBRM1 abundance or poor recovery of nuclear protein is possible. | Verify loading and transfer, then compare a nuclear fraction with a positive-control lysate. |
Experiment evidence
Western blot images showing PBRM1 band
A01130-1 · PBRM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PBRM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PBRM1 in Western blot, you can use cerebral cortex tissue, where HPA reports high expression.
Positive control: Cerebral cortex (IHC candidate; verify WB)
Negative control: Liver (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Because PBRM1 is nuclear, nuclear-enriched lysate may improve detection.
HPA tissue expression evidence for PBRM1
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 | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | hepatocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Prostate | glandular cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced PBRM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PBRM1, answered from its protein features.
How should PBRM1 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 PBRM1 isoforms could produce smaller bands?
Isoforms · UniProt lists nine isoforms. Isoform 6 lacks residues 857–1689 and could be substantially smaller. Several other isoforms lack shorter segments, including residues 1430–1484 in isoforms 2, 3, 4, 5, 7 and 9. These are UniProt canonical-sequence coordinates; confirm which isoforms your antibody recognizes before assigning bands.
Could phosphorylation affect the PBRM1 band pattern?
PTM · UniProt lists phosphoserines at 10, 39, 131, 178, 316, 319, 353, 355, 371, 375, 498, 509, 636, 648, 689, 948, 987, 1119, 1405 and 1453, plus phosphotyrosines at 134 and 1289. These are UniProt canonical-sequence coordinates. Compare samples with and without phosphatase treatment if phosphorylation is suspected; the annotations alone do not establish a visible shift.
UniProt lists N6-acetyllysine at residue 414 in canonical-sequence coordinates. If studying acetylated PBRM1, confirm that the antibody detects this site or use an appropriate enrichment strategy. The annotation does not establish that acetylation causes a resolvable band shift.
UniProt lists N6-acetyllysine at residue 414 in canonical-sequence coordinates. If studying acetylated PBRM1, confirm that the antibody detects this site or use an appropriate enrichment strategy. The annotation does not establish that acetylation causes a resolvable band shift.
Does this guide establish induction of PBRM1?
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 PBRM1 Western blot?
Transfer · PBRM1 is a large protein: its canonical predicted mass is 192.9 kDa, and the supplied apparent band is about 240 kDa. Choose transfer conditions that retain large proteins and check transfer at the high end of the molecular weight ladder. The UniProt features do not specify a membrane or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01130-1 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 PBRM1 bands be quantified?
Quantitation · Quantify a consistently identified PBRM1 band within the linear detection range, using the same band definition across samples. PBRM1 has nine annotated isoforms and numerous modification sites, so record whether additional bands are included. Do not combine them as total PBRM1 without establishing their identities.
Why might PBRM1 appear near 240 kDa instead of 192.9 kDa?
Interpretation · The supplied apparent band is about 240 kDa, while the canonical sequence predicts 192.9 kDa. Compare it with a molecular weight ladder and confirm its identity with a second antibody or PBRM1 depletion. The listed modifications and isoforms do not, by themselves, explain this difference.
Check antibody recognition and PBRM1 depletion, then compare band sizes with the annotated isoforms. Isoform 6 lacks residues 857–1689; isoforms 4 and 9 lack 989–1013, and isoform 4 also lacks 1336–1362. These are UniProt canonical-sequence coordinates. A matching size alone cannot identify an isoform or modification.
Check antibody recognition and PBRM1 depletion, then compare band sizes with the annotated isoforms. Isoform 6 lacks residues 857–1689; isoforms 4 and 9 lack 989–1013, and isoform 4 also lacks 1336–1362. These are UniProt canonical-sequence coordinates. A matching size alone cannot identify an isoform or modification.
Boster reagents
PBRM1 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-PBRM1 Antibody Picoband®
Cat # A01130-1
The catalog reports A01130-1, a rabbit polyclonal anti-PBRM1 antibody with reported human, mouse, and rat reactivity. Its Western blot image reports a band near 240 kDa, versus an expected 193 kDa; this discrepancy limits interpretation.
Which to pick: A01130-1 is the only listed option with a Western blot image. The caption documents human K562, HeLa, and U2OS; rat C6; and mouse brain and NIH/3T3 lysates. Choose based on your sample and assess the reported band size discrepancy.
Source: BosterBio PBRM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86U86.
- Human Protein Atlas. PBRM1 tissue expression.
- PMC11002309 — target-verified WB comparison
- PMC4599275 — target-verified WB comparison
- PMC9204009 — target-verified WB comparison