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- Table of Contents
PSMB8 / Proteasome subunit beta type-8 · Western blot design guide
Design a Western Blot for PSMB8
Real validated PSMB8 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PSMB8 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
PSMB8 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 | ~30.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Lung (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Cleaved | |
| Caveat | Propeptide processing | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated PSMB8 Western Blot Protocols
The M02188-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | U937 cell lysate (catalog M02188-2) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02188-2; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected PSMB8 Western Blot Band Size?
PSMB8 has a predicted 30.4 kDa precursor; propeptide cleavage and isoforms may affect migration, but no empirical band size or distinct isoform pattern is supplied.
| Band near 30.4 kDa | consistent with the predicted precursor, pending identity controls |
| Band below 30.4 kDa | could reflect removal of the residues 1–72 propeptide |
| Two bands at different positions | could reflect precursor and mature PSMB8 |
| Several discrete bands | could include isoforms 1 and 2, though their migration is unknown |
💡Expected PSMB8 appearanceUniProt predicts a 30.4 kDa precursor; removal of its propeptide could yield a smaller mature band, but no empirical band size is supplied, so confirm band identity with controls.
| Predicted precursor mass | 30.4 kDa is the UniProt prediction, not a measured band position |
| Propeptide at residues 1–72 | cleavage can produce a smaller mature protein; its apparent size is unknown |
| Isoform 1 | may differ in apparent size from isoform 2; direction and separation are unknown |
| Isoform 2 | may differ in apparent size from isoform 1; direction and separation are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PSMB8 may be below detection in the tested lysate | compare with U937 lysate as a positive control and check loading |
| Band higher than expected | the 30.4 kDa precursor may be present when a mature band was expected | compare precursor and mature forms using an appropriate band-identity control |
| Band lower than expected | the residues 1–72 propeptide may have been removed | check antibody recognition of mature PSMB8 and confirm identity |
| Multiple bands | precursor processing or isoforms 1 and 2 may contribute | confirm PSMB8 specificity with depletion and compare band patterns |
| Weak or no signal | PSMB8 may be below the assay detection limit | check loading, antibody performance and a U937 lysate control |
Experiment evidence
Western blot images showing PSMB8 band
M02188-2 · PSMB8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PSMB8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PSMB8 in Western blot, you can use lung tissue lysate.
Positive control: Lung (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Tissue lysates are suitable, with adipose tissue providing a potential negative control.
HPA tissue expression evidence for PSMB8
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 | alveolar cells type I | High | Protein (IHC) | HPA → |
| Skin | lymphocytes | High | 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 → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced PSMB8 Western Blot Tips
Deeper troubleshooting and optimisation questions for PSMB8, answered from its protein features.
How should PSMB8 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 PSMB8 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 has an alternative sequence at the N terminus, annotated as VAR_SEQ 1..49. Check whether the antibody target includes this region. Isoform annotation alone does not establish that both isoforms are expressed or resolve as separate bands.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of PSMB8?
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 PSMB8 Western blot?
Transfer · Plan to retain proteins around the 30.4 kDa precursor size and potentially smaller processed forms, then verify transfer with a size marker. The supplied UniProt features do not specify a membrane, transfer method, or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M02188-2 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 PSMB8 bands be quantified?
Quantitation · Define the band being measured before comparing samples. The annotated propeptide at UniProt residues 1..72 means precursor and processed PSMB8 could differ in size; the two annotated isoforms also have different N terminal sequences. Keep any separately resolved bands distinct unless their identities are verified.
Why might PSMB8 migrate below its predicted 30.4 kDa?
Interpretation · The 30.4 kDa prediction includes a propeptide at UniProt residues 1..72. Processing could produce a smaller mature protein, but these features do not establish its apparent mass or explain a particular band without experimental evidence.
PSMB8 is annotated as a zymogen with a propeptide at UniProt residues 1..72. A precursor and a processed form could differ in size. Check which form the antibody recognizes before assigning a band; the supplied features do not establish that both forms will be visible.
Compare their sizes with the 30.4 kDa precursor prediction and consider propeptide processing at UniProt residues 1..72 and the isoform 2 N terminal alternative sequence. Confirm band identity experimentally: these annotations alone do not assign an unexpected band, and no empirical apparent band is supplied.
PSMB8 is annotated as a zymogen with a propeptide at UniProt residues 1..72. A precursor and a processed form could differ in size. Check which form the antibody recognizes before assigning a band; the supplied features do not establish that both forms will be visible.
Compare their sizes with the 30.4 kDa precursor prediction and consider propeptide processing at UniProt residues 1..72 and the isoform 2 N terminal alternative sequence. Confirm band identity experimentally: these annotations alone do not assign an unexpected band, and no empirical apparent band is supplied.
Boster reagents
PSMB8 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-PSMB8 / LMP7 Monoclonal Antibody
Cat # M02188-2
The listed anti-PSMB8/LMP7 monoclonal antibody, M02188-2, has a Western blot image showing proteasome 20S LMP7 expression in U937 cell lysate. The product lists Human, Mouse, and Rat reactivity; the supplied image documents only the stated sample context.
Which to pick: M02188-2 is the only listed option. Its Western blot image uses U937 cell lysate; check that the listed reactivity and this demonstrated sample context fit your experiment.
Source: BosterBio PSMB8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P28062.
- Human Protein Atlas. PSMB8 tissue expression.
- PMC12390170 — target-verified WB comparison
- PMC12058196 — target-verified WB comparison
- PMC11896265 — target-verified WB comparison