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- Table of Contents
PSMB4 / Proteasome subunit beta type-4 · Western blot design guide
Design a Western Blot for PSMB4
Source-linked PSMB4 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PSMB4 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
PSMB4 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 | ~29.2 kDa | |
| Observed band | ~25 and 29 kDa | |
| Gel | 5–20% (catalog A05105-1) | |
| Positive control | Lung (IHC candidate; verify WB) +4 more | |
| Negative control | Cervix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Propeptide processing | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked PSMB4 Western Blot Protocol Options
The A05105-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human HepG2, human A431, human RT4, human U251, human placenta, human MCF-7, rat skeletal muscle, mouse liver, mouse skeletal muscle (catalog A05105-1) |
| Gel % | 5–20% (catalog A05105-1) |
| Load | 30 ug; reducing conditions (catalog A05105-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05105-1) |
| Membrane | nitrocellulose membrane (catalog A05105-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05105-1) |
| Primary antibody | A05105-1 · 0.5 μg/mL (catalog A05105-1) |
| Primary incubation | overnight at 4°C (catalog A05105-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05105-1) |
| Secondary incubation | 1.5 hour at RT (catalog A05105-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05105-1) |
| Detection | ECL (catalog A05105-1) |
Section 2
What Is the Expected PSMB4 Western Blot Band Size?
PSMB4 is predicted at 29.2 kDa, while blots show about 25 and 29 kDa; the cause of the difference is not established.
| Band near 29 kDa | Matches the 29.2 kDa full-length prediction; confirm identity with a control. |
| Band near 25 kDa | An empirically detected PSMB4 band; propeptide cleavage is possible but unconfirmed. |
| Bands near 25 and 29 kDa | Both reported bands are present; their relationship remains unestablished. |
| Lower band predominates over the 29 kDa band | Could reflect propeptide processing, but band identity requires validation. |
💡Expected PSMB4 appearanceUniProt predicts a 29.2 kDa precursor, while antibody blots show bands near 25 and 29 kDa; the propeptide may affect migration, but use band-identity controls before assigning either band to a processing state.
| UniProt predicted mass | Places full-length PSMB4 at 29.2 kDa. |
| Propeptide at residues 1–45 | Adds sequence to the full-length precursor relative to a cleaved product. |
| Full-length precursor retention | Could favor the higher band relative to processed PSMB4. |
| Propeptide cleavage | Could produce a smaller mature protein; its apparent mass is not established. |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The full-length precursor retains residues 1–45 relative to processed PSMB4. | Compare with the reported 29 kDa band and verify identity with a knockdown or knockout control. |
| Band lower than expected | Propeptide cleavage is possible, though the 25 kDa band's identity is unproven. | Compare with the reported 25 kDa band and verify identity with a knockdown or knockout control. |
| Multiple bands | Blots report bands near 25 and 29 kDa; their relationship is unresolved. | Check whether both bands respond to PSMB4 knockdown or knockout. |
| Weak or no signal | PSMB4 occurs in both cytoplasm and nucleus, so recovery may vary by fraction. | Check loading and extraction, then compare whole-cell and subcellular fractions. |
| Fragments below expected size | Sample proteolysis could generate smaller immunoreactive fragments. | Prepare fresh lysate with protease inhibitors and confirm band identity by knockdown or knockout. |
Experiment evidence
Western blot images showing PSMB4 band
A05105-1 · PSMB4
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PSMB4 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PSMB4 in Western blot, you can use lung tissue lysate, which HPA scores High.
Positive control: Lung (IHC candidate; verify WB)
Negative control: Cervix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Cervix is HPA Not detected, but confirm that your lysate gives no PSMB4 signal.
HPA tissue expression evidence for PSMB4
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 |
|---|---|---|---|---|
| Lung | macrophages | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PSMB4 Western Blot Tips
Deeper troubleshooting and optimisation questions for PSMB4, answered from its protein features.
How should PSMB4 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 PSMB4 isoforms explain the two observed bands?
Isoforms · The supplied features list one isoform and no alternative sequence. Isoforms therefore provide no supported explanation for the approximately 25 and 29 kDa bands. Consider propeptide processing when interpreting them.
Which reported modifications could affect PSMB4 band interpretation?
PTM · UniProt lists N-acetylmethionine at residue 1, phosphoserine at residue 26, and phosphotyrosine at residue 102. Residues 1 and 26 fall within the 1–45 propeptide; residue 102 lies outside it. These features alone do not demonstrate a visible shift or explain the measured band positions.
UniProt Ser26 is within the propeptide at residues 1–45, so cleavage would remove that site from mature PSMB4. Interpret a phospho-Ser26 signal with processing state in mind, and check that antibody residue numbering matches UniProt numbering.
UniProt Ser26 is within the propeptide at residues 1–45, so cleavage would remove that site from mature PSMB4. Interpret a phospho-Ser26 signal with processing state in mind, and check that antibody residue numbering matches UniProt numbering.
Does this guide establish induction of PSMB4?
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 PSMB4?
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 A05105-1 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 PSMB4 be quantified across cell compartments?
Quantitation · PSMB4 is listed in both cytoplasm and nucleus and can move into the nucleus after interacting with AKIRIN2. If comparing fractions, quantify nuclear and cytoplasmic signals separately; a change in one fraction may reflect redistribution rather than a change in total PSMB4.
Why might PSMB4 appear near both 25 and 29 kDa?
Interpretation · The predicted 29.2 kDa mass includes the UniProt propeptide at residues 1–45. Its removal would reduce the mass, consistent with a band near 25 kDa; a band near 29 kDa could represent uncleaved protein. Band position alone cannot establish either identity.
Compare it with the reported approximately 25 and 29 kDa bands and the 1–45 propeptide boundary. Check whether the antibody recognizes a region retained after cleavage. PSMB4 belongs to a proteasome complex, but complex membership alone does not identify an unexpected band.
Compare it with the reported approximately 25 and 29 kDa bands and the 1–45 propeptide boundary. Check whether the antibody recognizes a region retained after cleavage. PSMB4 belongs to a proteasome complex, but complex membership alone does not identify an unexpected band.
Boster reagents
PSMB4 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-PSMB4 Antibody Picoband®
Cat # A05105-1
A05105-1 is a rabbit polyclonal anti-PSMB4 antibody with a Western blot image showing bands near 25 and 29 kDa; the expected size is 29 kDa. The caption reports human, rat, and mouse samples. No publication evidence is supplied.
Which to pick: A05105-1 is the only listed option. Its reported reactivity is human, mouse, and rat, and its Western blot image includes human cell and tissue lysates, rat skeletal muscle, and mouse liver and skeletal muscle. Match your sample to those tested contexts.
Source: BosterBio PSMB4 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P28070.
- Human Protein Atlas. PSMB4 tissue expression.
- PMC13257190 — target-verified WB comparison
- PMC12062861 — target-verified WB comparison
- PMC11977064 — target-verified WB comparison