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PSMD1 / 26S proteasome non-ATPase regulatory subunit 1 · Western blot design guide
Design a Western Blot for PSMD1
Source-linked PSMD1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PSMD1 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
PSMD1 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 | ~105.8 kDa | |
| Observed band | ~106 kDa | |
| Gel | 8% (catalog A06293-1) | |
| Positive control | Breast (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked PSMD1 Western Blot Protocol Options
The A06293-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human HepG2, human PC-3, rat brain, rat liver, mouse brain, mouse liver (catalog A06293-1) |
| Gel % | 8% (catalog A06293-1) |
| Load | 30 ug; reducing conditions (catalog A06293-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06293-1) |
| Membrane | nitrocellulose membrane (catalog A06293-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06293-1) |
| Primary antibody | A06293-1 · 0.5 μg/mL (catalog A06293-1) |
| Primary incubation | overnight at 4°C (catalog A06293-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06293-1) |
| Secondary incubation | 1.5 hour at RT (catalog A06293-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06293-1) |
| Detection | ECL (catalog A06293-1) |
Section 2
What Is the Expected PSMD1 Western Blot Band Size?
PSMD1 is predicted at 105.8 kDa and observed at ~106 kDa; the cause of any apparent difference is not established.
| Band at ~106 kDa | Matches the empirical PSMD1 band and its 105.8 kDa predicted mass. |
| Single sharp band near 106 kDa | Consistent with detectable PSMD1; the supplied evidence does not establish separate isoform bands. |
| Additional band at a different size | Could represent isoform 1 or 2, but its identity needs validation. |
| Doublet near 106 kDa | Could involve the documented isoforms or modified PSMD1; separate migration is unproven. |
💡Expected PSMD1 appearancePSMD1 has a predicted mass of 105.8 kDa and an empirical band at ~106 kDa; confirm band identity with appropriate antibody controls if additional bands appear.
| Predicted PSMD1 mass | 105.8 kDa predicted; the empirical band is ~106 kDa. |
| Isoforms 1 and 2 | Different forms are documented, but their relative sizes and separate migration are not supplied. |
| Phosphothreonine at residue 273 | Modification is documented; no visible band shift is established. |
| N6-acetyllysine at residue 310 | Modification is documented; no visible band shift is established. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PSMD1 may be below detection in the prepared sample. | Check lysate loading and transfer with a control protein, then verify antibody performance with a positive lysate. |
| Band higher than expected | Identity or migration of the higher band is unestablished. | Check denaturation and compare with an independent PSMD1 antibody or knockdown control. |
| Band lower than expected | An isoform or breakdown product is possible, but neither has a supplied band size. | Use protease inhibitors and verify identity with an independent antibody or knockdown control. |
| Multiple bands | Isoforms 1 and 2 are documented, but distinct bands are unproven. | Compare isoform-specific evidence and use a PSMD1 knockdown control to identify specific bands. |
| Weak or no signal | The ~106 kDa PSMD1 band may fall below assay detection. | Check loading and transfer, then test a validated positive lysate. |
| Fragments below expected size | PSMD1 breakdown is possible; fragment sizes are not documented here. | Prepare fresh lysate with protease inhibitors and confirm fragment identity with a second antibody. |
Experiment evidence
Western blot images showing PSMD1 band
A06293-1 · PSMD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PSMD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PSMD1 in Western blot, you can use breast tissue, which shows high expression in HPA.
Positive control: Breast (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 Not-detected tissue is listed, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for PSMD1
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 |
|---|---|---|---|---|
| Breast | myoepithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Lung | alveolar cells | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced PSMD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for PSMD1, answered from its protein features.
How should PSMD1 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 PSMD1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 lacks residues 797–827 of the canonical sequence, so it could migrate differently. The features alone do not establish that a second band will be visible.
An antibody recognizing canonical residues 797–827 may miss isoform 2 because that segment is absent. Check the antibody’s stated epitope against this region when interpreting a missing or weaker isoform band.
An antibody recognizing canonical residues 797–827 may miss isoform 2 because that segment is absent. Check the antibody’s stated epitope against this region when interpreting a missing or weaker isoform band.
Which phosphorylation sites matter when interpreting PSMD1 bands?
PTM · UniProt lists phosphothreonine at 273, 311, and 830 and phosphoserine at 290, 315, and 834. These are canonical UniProt coordinates; numbering after the isoform 2 deletion can differ. Site annotation alone does not establish a resolvable band shift.
UniProt lists N-acetylmethionine at position 1 and N6-acetyllysine at positions 310 and 720, using canonical coordinates. These annotations do not establish a distinct Western blot band.
UniProt lists N-acetylmethionine at position 1 and N6-acetyllysine at positions 310 and 720, using canonical coordinates. These annotations do not establish a distinct Western blot band.
Does this guide establish induction of PSMD1?
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 PSMD1?
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 A06293-1 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 PSMD1 bands be quantified?
Quantitation · Use a consistent band assignment near the observed 106 kDa position across samples. If another band appears, verify its identity before combining its signal with the main band; UniProt lists two isoforms but does not establish which bands an antibody detects.
Why might PSMD1 migrate near 106 kDa?
Interpretation · The predicted mass is 105.8 kDa, close to the observed band near 106 kDa. The listed modifications do not by themselves establish a visible shift or explain any small difference between calculated and apparent mass.
Isoform 2 lacks canonical residues 797–827, making it a candidate for a lower band. Confirm its identity before assigning it to that isoform; the supplied features do not establish the apparent mass or identity of any additional band.
Isoform 2 lacks canonical residues 797–827, making it a candidate for a lower band. Confirm its identity before assigning it to that isoform; the supplied features do not establish the apparent mass or identity of any additional band.
Boster reagents
PSMD1 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-PSMD1 Antibody Picoband®
Cat # A06293-1
The catalog reports one anti-PSMD1 antibody for Western blot, A06293-1. Its supplied blot shows an approximately 106 kDa band in human cell and rat and mouse tissue lysates. Evidence here is limited to this product image.
Which to pick: A06293-1 is the only listed option. It is reported reactive with human, mouse, and rat, and its WB image includes A549, HepG2, PC-3, and rat and mouse brain and liver lysates under reducing conditions.
Source: BosterBio PSMD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99460.
- Human Protein Atlas. PSMD1 tissue expression.
- PMC12432587 — target-verified WB comparison
- PMC6039477 — target-verified WB comparison
- PMC7952820 — target-verified WB comparison