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- Table of Contents
LDB1 / LIM domain-binding protein 1 · Western blot design guide
Design a Western Blot for LDB1
Source-linked LDB1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LDB1 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
LDB1 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 | ~46.5 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bone marrow (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 + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked LDB1 Western Blot Protocol Options
The A01977 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01977; 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 | The membrane was washed and reacted with a 1:20,000 dilution of DyLight™800 conjugated Gt-a-Rabbit IgG [H&L] MX for 45 min at room temperature (800 nm channel, green) (catalog A01977) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected LDB1 Western Blot Band Size?
LDB1 has a predicted mass of 46.5 kDa; isoforms and homodimers could affect band patterns, but their migration has not been demonstrated here.
| Band near 46.5 kDa | Consistent with the predicted LDB1 mass; identity needs confirmation |
| Several discrete bands | Isoforms 1, 2, and 3 could contribute, but their migration is unknown |
| Band near twice the predicted mass | Could reflect a retained LDB1 homodimer; its migration is unverified |
| Faint band in whole-cell lysate | Nuclear localization may limit the signal in this sample |
💡Expected LDB1 appearanceUniProt predicts 46.5 kDa for LDB1; no empirical band size is supplied, so confirm a candidate band with an independent antibody or LDB1 depletion.
| Predicted LDB1 mass | Places the reference band near 46.5 kDa |
| Isoform 1 | May migrate differently from isoforms 2 and 3; relative size is unknown |
| Isoform 2 | May migrate differently from isoforms 1 and 3; relative size is unknown |
| Isoform 3 | May migrate differently from isoforms 1 and 2; relative size is unknown |
| Homodimer formation | Could yield a higher band if the dimer survives sample preparation |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear LDB1 may be poorly represented in the sampled lysate | Check a nuclear fraction and confirm recovery with a nuclear marker |
| Band higher than expected | A homodimer may survive sample preparation | Compare more strongly denatured samples and confirm identity by LDB1 depletion |
| Band lower than expected | An LDB1 splice isoform may migrate differently; its size is unknown | Confirm the band with an independent antibody or isoform-specific depletion |
| Multiple bands | Isoforms 1, 2, and 3 may contribute; their migration is unverified | Use LDB1 depletion to identify specific bands |
| Weak or no signal | Nuclear localization may reduce signal in whole-cell lysate | Enrich the nuclear fraction and check sample loading |
Experiment evidence
Western blot images showing LDB1 band
A01977 · LDB1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LDB1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LDB1 in Western blot, you can use bone marrow, an HPA high-expression tissue.
Positive control: Bone marrow (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 the samples.
⚠️Feasibility: LDB1 is nuclear, so nuclear enrichment may help if whole-tissue lysate gives a weak signal.
HPA tissue expression evidence for LDB1
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular 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 → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Salivary gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LDB1 Western Blot Tips
Deeper troubleshooting and optimisation questions for LDB1, answered from its protein features.
How should LDB1 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 LDB1 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Isoforms 2 and 3 lack canonical residues 1–36; isoform 2 also replaces residues 336–352 and lacks residues 353–411. These sequence differences could affect band position, but the features do not establish where any isoform runs.
Check the antibody epitope against the listed sequence changes. An epitope within canonical residues 1–36 is absent from isoforms 2 and 3; one within 353–411 is absent from isoform 2. The supplied features do not identify an antibody epitope.
Check the antibody epitope against the listed sequence changes. An epitope within canonical residues 1–36 is absent from isoforms 2 and 3; one within 353–411 is absent from isoform 2. The supplied features do not identify an antibody epitope.
Which LDB1 modifications matter when assessing bands?
PTM · The supplied UniProt coordinates list N-acetylserine at residue 2, phosphothreonine at 61, and phosphoserine at 265 and 302. These are canonical-sequence coordinates; antibody or paper numbering may differ. Their presence does not establish a visible shift.
Phosphorylation is listed at canonical residues Thr61, Ser265, and Ser302. Treat it as a possible explanation to test, rather than assigning an unexpected band to phosphorylation from its position. No observed band position is supplied.
Phosphorylation is listed at canonical residues Thr61, Ser265, and Ser302. Treat it as a possible explanation to test, rather than assigning an unexpected band to phosphorylation from its position. No observed band position is supplied.
Does this guide establish induction of LDB1?
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 LDB1?
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 A01977 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.
What should be considered when quantifying LDB1 bands?
Quantitation · Define whether the measurement targets one isoform or all detected isoforms. The three listed isoforms differ in sequence, so an antibody epitope may determine which contribute to the signal. Report the bands measured and keep the same band selection across samples.
Should LDB1 run at exactly 46.5 kDa?
Interpretation · 46.5 kDa is the predicted mass, and no observed band position is supplied. Compare the band with the predicted mass, but do not assign a difference to modification or splicing from position alone.
Compare them with the three listed isoforms and the antibody epitope. Isoforms 2 and 3 lack canonical residues 1–36, while isoform 2 also has changes from residue 336 onward. The supplied features alone cannot identify an additional band or establish that a modification caused it.
Compare them with the three listed isoforms and the antibody epitope. Isoforms 2 and 3 lack canonical residues 1–36, while isoform 2 also has changes from residue 336 onward. The supplied features alone cannot identify an additional band or establish that a modification caused it.
Boster reagents
LDB1 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-LIM domain-binding protein 1 LDB1 Antibody
Cat # A01977
A01977 is an anti-LDB1 antibody listed for human reactivity. Its Western blot image shows LDB1 detection in Jurkat whole cell lysate, with approximately 30 µg loaded and primary antibody diluted 1:1,800. The supplied evidence covers this illustrated context.
Which to pick: A01977 is the only listed option for LDB1. It has a Western blot image using Jurkat whole cell lysate; use the reported sample and conditions to judge its fit for your experiment.
Source: BosterBio LDB1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q86U70.
- Human Protein Atlas. LDB1 tissue expression.
- PMC3375123 — target-verified WB comparison
- PMC8770319 — target-verified WB comparison