This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
LAMB1 / Laminin subunit beta-1 · Western blot design guide
Design a Western Blot for LAMB1
Real validated LAMB1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LAMB1 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
LAMB1 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 | ~198 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Appendix (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 | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated LAMB1 Western Blot Protocols
The M03894 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | A431 cell lysate (catalog M03894) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03894; 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 LAMB1 Western Blot Band Size?
LAMB1 has a predicted 198 kDa precursor; cleavage, N-linked glycosylation, and disulfide-linked chain association may affect migration, but no empirical band size is supplied.
| Band near 198 kDa | compatible with the predicted precursor mass; identity needs confirmation |
| Band below 198 kDa | could reflect signal peptide removal, although mature migration is unknown |
| Band above 198 kDa | could reflect N-linked glycosylation; the size effect is unmeasured |
| Very high molecular mass band under nonreducing conditions | could reflect disulfide-linked association with laminin alpha and gamma chains |
| Little or no band in whole-cell lysate | consistent with secretion into the extracellular matrix |
💡Expected LAMB1 appearanceUniProt predicts a 198 kDa precursor, but no empirical band size is supplied; signal peptide cleavage, N-linked glycosylation, and chain association may affect migration, so confirm band identity with appropriate controls.
| Predicted precursor mass | provides a 198 kDa sequence-based reference, not a measured band |
| N-linked glycosylation at Asn120 and Asn356 | may alter apparent mass, with no measured shift supplied |
| N-linked glycosylation at Asn519 and Asn677 | may alter apparent mass, with no measured shift supplied |
| Signal peptide at residues 1–21 | cleavage makes the mature polypeptide smaller than the precursor |
| Disulfide-linked laminin alpha, beta, and gamma chains | may retain LAMB1 in a higher molecular mass complex under nonreducing conditions |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | LAMB1 is secreted into the extracellular matrix | Check conditioned medium or extracellular matrix extracts alongside lysate |
| Band higher than expected | Possible N-linked glycosylation or retained disulfide-linked chains | Compare deglycosylated and untreated samples under reducing and nonreducing conditions |
| Band lower than expected | Signal peptide removal may reduce polypeptide mass | Check antibody epitope coverage and confirm identity with an independent antibody |
| Broad smear instead of sharp band | Possible variation in N-linked glycosylation | Compare untreated material with a PNGase F-treated aliquot |
| Multiple bands | Possible differences in glycosylation or disulfide-linked association | Compare reducing conditions and PNGase F treatment, then verify bands with an independent antibody |
| Weak or no signal | Extracellular localization may limit recovery from cell lysate | Enrich the extracellular fraction and include a positive control |
Experiment evidence
Western blot images showing LAMB1 band
M03894 · LAMB1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LAMB1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LAMB1 in Western blot, you can use appendix tissue, which shows high expression in HPA.
Positive control: Appendix (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: LAMB1 is secreted into the extracellular matrix, so whole-cell lysates may give weak signal; use matrix-enriched material.
HPA tissue expression evidence for LAMB1
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 |
|---|---|---|---|---|
| Appendix | enterocytes | High | Protein (IHC) | HPA → |
| Colon | enterocytes | High | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Rectum | endothelial cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LAMB1 Western Blot Tips
Deeper troubleshooting and optimisation questions for LAMB1, answered from its protein features.
How should LAMB1 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 multiple LAMB1 bands represent annotated isoforms?
Isoforms · The supplied record lists one isoform and no alternative sequence. Do not assign additional bands to specific LAMB1 isoforms from this evidence alone.
Which LAMB1 glycosylation sites matter when interpreting bands?
PTM · The annotated N-linked sites are Asn120, 356, 519, 677, 1041, 1195, 1279, 1336, 1343, 1487, and 1542. Compare samples prepared under the same conditions when assessing mobility; the site annotations alone do not predict a band size.
UniProt lists phosphoserine at residue 250 and FAM20C-associated phosphoserine at residues 1478, 1496, 1666, and 1682. These are UniProt coordinates; paper or antibody numbering may differ. The annotations do not establish a visible shift on a Western blot.
UniProt lists phosphoserine at residue 250 and FAM20C-associated phosphoserine at residues 1478, 1496, 1666, and 1682. These are UniProt coordinates; paper or antibody numbering may differ. The annotations do not establish a visible shift on a Western blot.
Does this guide establish induction of LAMB1?
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 LAMB1?
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 M03894 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 LAMB1 bands be compared for quantitation?
Quantitation · Compare the same sample fraction and preparation conditions across samples. LAMB1 is a secreted extracellular matrix protein, so a cellular fraction and an extracellular fraction may contain different amounts. The supplied features provide no basis for assigning a particular band as the quantitative species.
Why might LAMB1 migrate differently from its predicted 198 kDa mass?
Interpretation · LAMB1 has a signal peptide at residues 1–21 and 11 annotated N-linked glycosylation sites. These features can affect the mature protein and its apparent mobility, but they do not establish a particular band shift. No observed band size is supplied.
LAMB1 is secreted and annotated in the extracellular space, extracellular matrix, and basement membrane. Include the relevant extracellular or matrix fraction when designing the blot, and compare like fractions across samples.
LAMB1 has 54 annotated disulfide bonds and belongs to a laminin complex containing alpha, beta, and gamma chains linked by disulfide bonds. Its signal peptide and glycosylation sites also warrant consideration. These features suggest possible influences on migration but cannot identify an unexpected band without further evidence.
LAMB1 is secreted and annotated in the extracellular space, extracellular matrix, and basement membrane. Include the relevant extracellular or matrix fraction when designing the blot, and compare like fractions across samples.
LAMB1 has 54 annotated disulfide bonds and belongs to a laminin complex containing alpha, beta, and gamma chains linked by disulfide bonds. Its signal peptide and glycosylation sites also warrant consideration. These features suggest possible influences on migration but cannot identify an unexpected band without further evidence.
Boster reagents
LAMB1 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-Laminin beta 1 LAMB1 Rabbit Monoclonal Antibody
Cat # M03894
The listed anti-LAMB1 antibody, M03894, is a rabbit monoclonal with reported Human, Mouse, and Rat reactivity. Its Western blot image shows Laminin beta 1 expression in A431 cell lysate; no additional WB sample validation is supplied.
Which to pick: M03894 is the only listed option. Its A431 cell lysate Western blot image provides a tested context; the reported Human, Mouse, and Rat reactivity does not establish WB performance in every sample from those species.
Source: BosterBio LAMB1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P07942.
- Human Protein Atlas. LAMB1 tissue expression.
- PMC11806178 — target-verified WB comparison
- PMC11808599 — target-verified WB comparison
- PMC5702717 — target-verified WB comparison