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- Table of Contents
PSMB1 / Proteasome subunit beta type-1 · Western blot design guide
Design a Western Blot for PSMB1
Source-linked PSMB1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PSMB1 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
PSMB1 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 | ~26.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 | Glycosylated + Phosphorylated | |
| Caveat | Precursor processing | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked PSMB1 Western Blot Protocol Options
The A06891 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A06891) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A06891) |
| Primary antibody | A06891 · 1:1000 (catalog A06891) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A06891) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A06891) |
Section 2
What Is the Expected PSMB1 Western Blot Band Size?
PSMB1 has a predicted 26.5 kDa precursor; propeptide cleavage and O-linked GlcNAc are annotated, but their effects on observed migration are not established.
| Band near 26.5 kDa | consistent with the predicted PSMB1 precursor; confirm identity with an antibody control |
| Band below 26.5 kDa | could reflect removal of the 1..28 propeptide |
| Single band without a resolved glycosylated form | O-linked GlcNAc is annotated at Ser58 and Ser209, but visible separation is not established |
| Bands in cytoplasmic and nuclear fractions | consistent with PSMB1 localization in both compartments |
💡Expected PSMB1 appearanceThe predicted 26.5 kDa mass describes the precursor; cleavage of its 1..28 propeptide may yield a smaller band, but no empirical band size or demonstrated migration effect is supplied, so confirm band identity experimentally.
| Predicted precursor mass | 26.5 kDa is the sequence-based reference, not a measured band position |
| Propeptide at residues 1..28 | cleavage can make the mature protein smaller than the precursor |
| O-linked GlcNAc at Ser58 | a modification site is annotated, but a visible shift is not established |
| O-linked GlcNAc at Ser209 | a modification site is annotated, but a visible shift is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | uncleaved precursor may run above a processed species | check precursor and mature forms with an independent PSMB1 antibody |
| Band lower than expected | removal of the 1..28 propeptide may reduce size | confirm the band with PSMB1 depletion and an independent antibody |
| Multiple bands | precursor and processed PSMB1 may coexist; other bands may be nonspecific | compare reducing blots after PSMB1 depletion |
| Weak or no signal | PSMB1 may be distributed between cytoplasm and nucleus | check matched fractions and whole-cell lysate with loading controls |
| Fragments below expected size | additional breakdown is not explained by the annotated propeptide alone | prepare lysate with protease inhibitors and verify fragments by PSMB1 depletion |
Experiment evidence
Western blot images showing PSMB1 band
A06891 · PSMB1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PSMB1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PSMB1 in Western blot, you can use adrenal gland lysate, which shows 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 such as Ponceau alongside.
⚠️Feasibility: HPA reports low expression even in the weakest tissues listed, so use siRNA knockdown or a KO line for a clean negative control.
HPA tissue expression evidence for PSMB1
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 → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Low | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PSMB1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PSMB1, answered from its protein features.
How should PSMB1 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 extra PSMB1 bands represent isoforms?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign extra bands to splice isoforms from this evidence alone; compare them with the expected precursor and processed forms and verify antibody specificity.
Which phosphorylation sites matter when interpreting PSMB1 bands?
PTM · UniProt lists phosphoserine at positions 62, 68, and 162, and phosphotyrosine at 150. These are coordinates in the supplied 241-residue sequence; antibody or paper numbering may differ. Phosphorylation is a plausible variable to investigate, but listed sites do not establish a visible shift.
UniProt lists O-linked GlcNAc at serines 58 and 209, using coordinates in the supplied sequence. Consider these sites when investigating migration differences, but their presence alone does not establish a visible shift or explain a particular band.
UniProt lists N-acetylmethionine at position 1 and N6-acetyllysine at position 204. Position 1 lies within the annotated propeptide at positions 1–28, so an assay for that N-terminal modification may distinguish precursor from processed protein. All positions use the supplied UniProt sequence numbering.
UniProt lists O-linked GlcNAc at serines 58 and 209, using coordinates in the supplied sequence. Consider these sites when investigating migration differences, but their presence alone does not establish a visible shift or explain a particular band.
UniProt lists N-acetylmethionine at position 1 and N6-acetyllysine at position 204. Position 1 lies within the annotated propeptide at positions 1–28, so an assay for that N-terminal modification may distinguish precursor from processed protein. All positions use the supplied UniProt sequence numbering.
Does this guide establish induction of PSMB1?
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 PSMB1?
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 A06891 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 nuclear and cytoplasmic PSMB1 signals be quantified?
Quantitation · PSMB1 is listed in both cytoplasm and nucleus, with nuclear translocation following interaction with AKIRIN2 and IPO9. Quantify each fraction with appropriate fraction loading controls and assess their relative distribution; a stronger nuclear signal need not mean more total PSMB1.
Why might PSMB1 migrate below its predicted 26.5 kDa mass?
Interpretation · The 26.5 kDa prediction is for the 241-residue sequence, which includes a propeptide at UniProt positions 1–28. Processing could yield a smaller mature protein. No observed band mass is supplied, so the size difference cannot be confirmed here.
Compare its size with the 26.5 kDa full-length prediction and account for the propeptide at UniProt positions 1–28. Check antibody specificity and whether the band changes with sample preparation. The listed phosphorylation, O-GlcNAc, and acetylation sites suggest variables to investigate, but none proves the identity of an unexpected band.
Compare its size with the 26.5 kDa full-length prediction and account for the propeptide at UniProt positions 1–28. Check antibody specificity and whether the band changes with sample preparation. The listed phosphorylation, O-GlcNAc, and acetylation sites suggest variables to investigate, but none proves the identity of an unexpected band.
Boster reagents
PSMB1 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-Proteasome subunit beta type-1 PSMB1 Antibody
Cat # A06891
the supplier A06891 is listed for human, mouse, and rat PSMB1 and has a Western blot image using cell-line extracts at 1:1000 dilution and 25 µg per lane. The caption does not identify the cell lines or their species.
Which to pick: A06891 is the only listed option. It has a WB image; check that its stated human, mouse, or rat reactivity fits your sample. The image caption does not establish which species were tested.
Source: BosterBio PSMB1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P20618.
- Human Protein Atlas. PSMB1 tissue expression.
- PMC10691970 — target-verified WB comparison
- PMC6409894 — target-verified WB comparison
- PMC13376088 — target-verified WB comparison