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- Table of Contents
LMNB2 / Lamin-B2 · Western blot design guide
Design a Western Blot for LMNB2
Source-linked LMNB2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LMNB2 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
LMNB2 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 | ~69.9 kDa | |
| Observed band | ~70 kDa | |
| Gel | 5–20% (catalog PA1636) | |
| Positive control | Caudate (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 | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked LMNB2 Western Blot Protocol Options
The PA1636 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human Hela (nucleus), human K562 (nucleus), human PC-3, rat C6, mouse NIH/3T3 (catalog PA1636) |
| Gel % | 5–20% (catalog PA1636) |
| Load | 30 ug; reducing conditions (catalog PA1636) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog PA1636) |
| Membrane | nitrocellulose membrane (catalog PA1636) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog PA1636) |
| Primary antibody | PA1636 · 0.5 μg/mL (catalog PA1636) |
| Primary incubation | overnight at 4°C (catalog PA1636) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog PA1636) |
| Secondary incubation | 1.5 hour at RT (catalog PA1636) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog PA1636) |
| Detection | ECL (catalog PA1636) |
Section 2
What Is the Expected LMNB2 Western Blot Band Size?
LMNB2 is predicted at 69.9 kDa and observed at ~70 kDa; the cause of their small difference is not established.
| Band at ~70 kDa | Matches the empirical LMNB2 band in the supplied blot. |
| Faint band in whole-cell lysate but stronger band in nuclear lysate | Consistent with LMNB2 localization at the nuclear lamina. |
| Band slightly below ~70 kDa | Could reflect removal of the 618–620 propeptide; a visible shift is unproven. |
| Higher band near twice the monomer size | Could represent a dimer that persists during electrophoresis; confirm its identity. |
💡Expected LMNB2 appearanceLMNB2 has a predicted mass of 69.9 kDa and an empirical band at ~70 kDa; confirm band identity with nuclear enrichment and an appropriate antibody control.
| Predicted LMNB2 mass | 69.9 kDa provides the monomer reference; the empirical band is ~70 kDa. |
| O-linked GlcNAc at Thr413 | Adds one monosaccharide; a resolvable shift is not established. |
| Propeptide at residues 618–620 | Its removal makes mature LMNB2 slightly smaller than the precursor; visible separation is unproven. |
| Dimer formation by similarity | Could produce a higher band if dimers persist during electrophoresis; no inter-chain disulfide is listed. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | LMNB2 is localized at the nuclear lamina and may be poorly recovered in the sampled lysate. | Check a nuclear fraction and verify extraction with a nuclear marker. |
| Band higher than expected | A lamin dimer or assembly could persist during sample preparation; its identity is unconfirmed. | Check denaturation and compare with the ~70 kDa band using a second LMNB2 antibody. |
| Band lower than expected | Propeptide removal changes the mass slightly; a substantially lower band has no established feature-based explanation. | Check sample integrity and confirm identity with a second antibody. |
| Multiple bands | The supplied features do not establish distinct resolvable LMNB2 species. | Compare nuclear enrichment and confirm candidate bands with a second antibody. |
| Weak or no signal | Nuclear-lamina LMNB2 may be under-recovered during extraction. | Assess nuclear extraction and loading with a nuclear marker. |
| Fragments below expected size | The listed three-residue propeptide does not explain substantially smaller fragments. | Check sample handling and confirm smaller bands with an independent LMNB2 antibody. |
Experiment evidence
Western blot images showing LMNB2 band
PA1636 · LMNB2
M05348 · LMNB2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LMNB2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LMNB2 in Western blot, you can use caudate tissue, which has high HPA expression.
Positive control: Caudate (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 tissue is listed as not detected, so use siRNA knockdown or a KO line for a clean negative control.
HPA tissue expression evidence for LMNB2
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 |
|---|---|---|---|---|
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
| Colon | endothelial cells | Low | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Low | Protein (IHC) | HPA → |
| Placenta | decidual cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced LMNB2 Western Blot Tips
Deeper troubleshooting and optimisation questions for LMNB2, answered from its protein features.
How should LMNB2 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.
Should LMNB2 isoforms produce separate bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore gives no basis for assigning separate bands to LMNB2 isoforms.
Which LMNB2 phosphorylation sites matter when interpreting bands?
PTM · UniProt lists phosphothreonine at 23 and 34, and phosphoserine at 37, 316, 407, 420, 422, 424, 426 and 497. These are UniProt coordinates; check the numbering convention of any antibody or paper before comparing sites. Site annotations alone do not establish a visible band shift.
Yes. UniProt lists O-linked GlcNAc on threonine 413. Use UniProt numbering when comparing this site with antibody or paper annotations. The site’s presence alone does not show that glycosylation shifts the observed band.
Yes. UniProt lists O-linked GlcNAc on threonine 413. Use UniProt numbering when comparing this site with antibody or paper annotations. The site’s presence alone does not show that glycosylation shifts the observed band.
Does this guide establish induction of LMNB2?
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 LMNB2?
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 PA1636 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.
Which band should be quantified for LMNB2?
Quantitation · Quantify the approximately 70 kDa band that matches both the supplied observation and the 69.9 kDa prediction. LMNB2 is annotated at the nuclear lamina, so compare like-for-like nuclear samples. Assess additional bands separately rather than adding them to the main signal without identifying them.
How does the observed LMNB2 band compare with its predicted mass?
Interpretation · LMNB2 has a predicted mass of 69.9 kDa, close to the supplied observed band at approximately 70 kDa. Its listed modifications do not, by themselves, establish a visible shift or explain any difference between calculated and apparent mass.
The record lists a propeptide at residues 618–620 and a cysteine methyl ester at residue 617, using UniProt coordinates. Keep those positions explicit when interpreting a C-terminal antibody’s target region; the annotations alone do not predict a resolvable band shift.
LMNB2 is described as forming dimers and larger assemblies by similarity. That makes assembly relevant to investigating a higher band, but the record does not show that an assembly survives Western blot preparation. Do not assign a higher band to LMNB2 from size alone.
The record lists a propeptide at residues 618–620 and a cysteine methyl ester at residue 617, using UniProt coordinates. Keep those positions explicit when interpreting a C-terminal antibody’s target region; the annotations alone do not predict a resolvable band shift.
LMNB2 is described as forming dimers and larger assemblies by similarity. That makes assembly relevant to investigating a higher band, but the record does not show that an assembly survives Western blot preparation. Do not assign a higher band to LMNB2 from size alone.
Boster reagents
LMNB2 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-Lamin B2/LMNB2 Antibody Picoband®
Cat # PA1636
Real WB data
Anti-Lamin B2 Monoclonal Antibody
Cat # M05348
The catalog reports two anti-LMNB2 antibodies for Western blotting: PA1636 (polyclonal) and M05348 (monoclonal). Both list human, mouse, and rat reactivity and show bands near 70 kDa in product blot images. No independent validation is supplied.
Which to pick: Both have WB images. PA1636 includes human whole-cell and nuclear lysates, rat C6, and mouse NIH/3T3; M05348 shows human, rat, and mouse whole-cell lysates. Choose based on your sample type and preference for polyclonal or monoclonal format.
Source: BosterBio LMNB2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q03252.
- Human Protein Atlas. LMNB2 tissue expression.
- PMC11810055 — target-verified WB comparison
- PMC7851573 — target-verified WB comparison
- PMC7846963 — target-verified WB comparison