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- Table of Contents
MZB1 / Marginal zone B- and B1-cell-specific protein · Western blot design guide
Design a Western Blot for MZB1
Source-linked MZB1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MZB1 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
MZB1 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 | ~20.7 kDa | |
| Observed band | ~19 kDa | |
| Gel | 5–20% (catalog A08281-1) | |
| Positive control | Rectum (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 | Cleaved | |
| Caveat | Signal peptide cleavage | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked MZB1 Western Blot Protocol Options
The A08281-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Raji, human Jurkat (catalog A08281-1) |
| Gel % | 5–20% (catalog A08281-1) |
| Load | 30 ug; reducing conditions (catalog A08281-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A08281-1) |
| Membrane | nitrocellulose membrane (catalog A08281-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A08281-1) |
| Primary antibody | A08281-1 · 0.25 μg/mL (catalog A08281-1) |
| Primary incubation | overnight at 4°C (catalog A08281-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A08281-1) |
| Secondary incubation | 1.5 hour at RT (catalog A08281-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A08281-1) |
| Detection | ECL (catalog A08281-1) |
Section 2
What Is the Expected MZB1 Western Blot Band Size?
MZB1 is predicted at 20.7 kDa and observed near 19 kDa; the cause of the difference is not established.
| Band near 19 kDa | Empirical MZB1 band; confirm identity with antibody and sample controls |
| Band near 20.7 kDa | Compatible with the predicted full-length precursor mass |
| Band below the precursor position | Could reflect signal-peptide cleavage; its migration is not established |
| Several discrete bands | Could reflect isoforms 1–5; their individual migration is not supplied |
| Little or no band in whole-cell lysate | Isoform 1 can be secreted, though it also resides in the ER lumen |
💡Expected MZB1 appearanceMZB1 has a predicted full-length mass of 20.7 kDa and an empirical band near 19 kDa; the difference is unexplained by the supplied evidence, so confirm identity with appropriate controls.
| Predicted full-length mass | 20.7 kDa is the sequence-based precursor reference; the empirical band is near 19 kDa |
| Signal peptide at residues 1–22 | Cleavage could yield a smaller mature protein, but its apparent mass is not supplied |
| Isoforms 1, 2 and 3 | Alternative splicing could alter migration; individual sizes and ordering are unknown |
| Isoforms 4 and 5 | Alternative splicing could alter migration; individual sizes and ordering are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Isoform 1 can be secreted, reducing its abundance in whole-cell lysate | Check conditioned medium alongside lysate and include a suitable positive control |
| Band higher than expected | An uncleaved precursor is possible; the band identity is unestablished | Compare with the 20.7 kDa precursor reference and verify antibody specificity |
| Band lower than expected | Signal-peptide cleavage is possible, but it does not establish the cause of the 19 kDa band | Compare with the empirical 19 kDa band and confirm identity using an independent antibody or depletion control |
| Multiple bands | Five annotated isoforms could contribute, but distinct migration is unproven | Compare isoform-specific samples or reagents and verify bands with depletion controls |
| Weak or no signal | Secretion or low abundance in the tested lysate may limit detection | Check conditioned medium, sample expression and a positive-control lysate |
Experiment evidence
Western blot images showing MZB1 band
A08281-1 · MZB1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MZB1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MZB1 in Western blot, you can use rectum tissue, which has high HPA expression.
Positive control: Rectum (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside samples.
⚠️Feasibility: Because MZB1 is secreted, whole-cell lysates may give weak signal; consider conditioned medium.
HPA tissue expression evidence for MZB1
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 |
|---|---|---|---|---|
| Rectum | mucosal lymphoid cells | High | Protein (IHC) | HPA → |
| Appendix | germinal center cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Colon | mucosal lymphoid cells | Medium | Protein (IHC) | HPA → |
| Lymph node | germinal center 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MZB1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MZB1, answered from its protein features.
How should MZB1 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.
How could MZB1 isoforms affect band interpretation?
Isoforms · UniProt lists five isoforms with substantial sequence differences. Isoform 3 lacks positions 1–92; isoforms 4 and 5 lack much of the C-terminal sequence. Check whether the antibody epitope is retained in each isoform before assigning bands by size.
Are glycosylation or other modified residues listed for MZB1?
PTM · The supplied UniProt features list no glycosylation sites or modified residues. Do not attribute a band shift to a specific modification based on these features.
Does this guide establish induction of MZB1?
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 MZB1?
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 A08281-1 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 MZB1 bands be quantified across samples?
Quantitation · Quantify the same identified band across matched samples. UniProt places isoform 1 in the ER lumen and also lists it as secreted, so keep cellular and secreted fractions distinct when comparing abundance.
Why might MZB1 run near 19 kDa instead of 20.7 kDa?
Interpretation · The 20.7 kDa prediction is for the full-length protein, which has a signal peptide at UniProt positions 1–22. Signal peptide removal could contribute to a lower apparent mass, but these features alone do not establish the cause of the observed ~19 kDa band.
UniProt lists three disulfide bonds. Compare reducing and nonreducing lanes if migration changes are in question. Disulfide bond presence alone does not predict a visible shift or explain the ~19 kDa band.
Check the antibody epitope against the five listed isoforms, especially the large missing regions in isoforms 3, 4, and 5. Compare bands across matched reducing conditions and sample fractions. Size alone cannot establish which isoform or processing event produced a band.
UniProt lists three disulfide bonds. Compare reducing and nonreducing lanes if migration changes are in question. Disulfide bond presence alone does not predict a visible shift or explain the ~19 kDa band.
Check the antibody epitope against the five listed isoforms, especially the large missing regions in isoforms 3, 4, and 5. Compare bands across matched reducing conditions and sample fractions. Size alone cannot establish which isoform or processing event produced a band.
Boster reagents
MZB1 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-MZB1 Antibody Picoband®
Cat # A08281-1
The catalog reports one human-reactive anti-MZB1 antibody, A08281-1, with a Western blot image of human Raji and Jurkat whole-cell lysates. The caption reports a band near 19 kDa versus an expected 21 kDa. Evidence is limited to the supplied product validation.
Which to pick: A08281-1 is the only listed option. Its WB image documents testing in human Raji and Jurkat lysates under reducing conditions, using 0.25 μg/mL primary antibody. Confirm performance in your own sample type and conditions.
Source: BosterBio MZB1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8WU39.
- Human Protein Atlas. MZB1 tissue expression.
- PMC12966909 — target-verified WB comparison
- PMC7736851 — target-verified WB comparison
- PMC4052763 — target-verified WB comparison