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- Table of Contents
LIMA1 / LIM domain and actin-binding protein 1 · Western blot design guide
Design a Western Blot for LIMA1
Source-linked LIMA1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-LIMA1 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
LIMA1 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 | ~85.2 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 more | |
| Negative control | Caudate (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked LIMA1 Western Blot Protocol Options
The A05231 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human colon tissue lysate (catalog A05231) |
| Gel % | 8–10% (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 | A05231 · (A) 0.5 and (B) 1 μg/ml (catalog A05231) |
| 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 LIMA1 Western Blot Band Size?
LIMA1 has a predicted mass of 85.2 kDa; isoforms could affect band patterns, but no empirical migration is supplied.
| Band near 85.2 kDa | Consistent with the predicted LIMA1 mass; confirm identity with controls |
| Several bands at different positions | Could reflect the five annotated isoforms; their migration is unverified |
| Close doublet | Could reflect isoforms or phosphorylation; neither pattern is established |
| Band enriched in a cytoskeletal fraction | Consistent with LIMA1's cytoskeletal localization |
💡Expected LIMA1 appearanceUniProt predicts 85.2 kDa for LIMA1, but no empirical band size is supplied; confirm any candidate band with antibody and depletion controls because isoform migration is unknown.
| UniProt predicted mass | Provides an 85.2 kDa reference, not a measured band position |
| Beta and Alpha isoforms | May differ in apparent size; individual masses are unavailable |
| Isoform 3 | May differ in apparent size; its mass is unavailable |
| Isoforms 4 and 5 | May differ in apparent size; individual masses are unavailable |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoskeleton-associated LIMA1 may be poorly recovered during extraction | Check insoluble fractions and compare extraction conditions |
| Band higher than expected | An isoform or altered migration is possible; the cause is unestablished | Compare antibody reagents and use LIMA1 depletion to test identity |
| Band lower than expected | An isoform is possible; no isoform mass is supplied | Confirm identity with LIMA1 depletion and an antibody to another epitope |
| Multiple bands | Five isoforms are annotated, but their band positions are unknown | Compare isoform expression and test which bands disappear after LIMA1 depletion |
| Weak or no signal | Extraction may underrecover cytoskeleton-associated LIMA1 | Check fraction recovery and include a validated positive lysate |
Experiment evidence
Western blot images showing LIMA1 band
A05231 · LIMA1
A05231-1 · LIMA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for LIMA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for LIMA1 in Western blot, you can use appendix tissue, which shows high expression in HPA.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Caudate (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Intracellular localization and an HPA not-detected tissue make tissue-based controls feasible.
HPA tissue expression evidence for LIMA1
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 | endocrine cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced LIMA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for LIMA1, answered from its protein features.
How should LIMA1 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 LIMA1 isoforms produce different bands?
Isoforms · Yes. Five isoforms are listed. Isoform 3 lacks residues 1–302; Alpha and 5 lack residues 1–160. Compare each band with the isoforms your antibody can recognize before assigning it. The supplied features do not establish their apparent band positions.
Yes. An antibody recognizing residues 1–160 would miss Alpha and isoform 5, while one recognizing residues 1–302 would miss isoform 3. Check the antibody's stated target region against these UniProt sequence changes before comparing isoform bands.
Yes. An antibody recognizing residues 1–160 would miss Alpha and isoform 5, while one recognizing residues 1–302 would miss isoform 3. Check the antibody's stated target region against these UniProt sequence changes before comparing isoform bands.
Could phosphorylation explain a shifted LIMA1 band?
PTM · UniProt lists phosphorylation, including Ser225, Tyr229, Ser230 and Ser601/604/609/617. These sites make phosphorylation worth considering when comparing bands across samples, but their presence alone does not establish a visible shift or explain a difference from 85.2 kDa. Keep UniProt numbering distinct from any antibody or paper numbering.
Does this guide establish induction of LIMA1?
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 LIMA1?
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 A05231 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 I quantify LIMA1 across samples?
Quantitation · Define which band you will measure and use the same band definition across samples. Five isoforms are listed, and LIMA1 is annotated in the cytoplasm, cytoskeleton, focal adhesions and at the cell membrane. Keep the sampled fraction consistent when comparing abundance.
What size band should I expect for LIMA1?
Interpretation · The supplied 759-residue sequence has a predicted mass of 85.2 kDa. No observed Western blot band is provided, so use 85.2 kDa as a starting reference, not an established apparent mass.
Check whether an extra band is compatible with the listed isoforms: isoform 3 lacks residues 1–302, Alpha and 5 lack 1–160, and isoforms 4 and 5 have an R-to-PG change at UniProt position 344. The features alone cannot identify an observed band or prove that a modification caused it.
Check whether an extra band is compatible with the listed isoforms: isoform 3 lacks residues 1–302, Alpha and 5 lack 1–160, and isoforms 4 and 5 have an R-to-PG change at UniProt position 344. The features alone cannot identify an observed band or prove that a modification caused it.
Boster reagents
LIMA1 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-LIMA1 Antibody
Cat # A05231
Real WB data
Anti-EPLIN/LIMA1 Antibody Picoband®
Cat # A05231-1
Both listed anti-LIMA1 antibodies are reported as human-reactive and have Western blot images. A05231 was tested on human colon tissue lysate; A05231-1 was tested on four human cell lines, with reported bands near 85 and 100 kDa. Evidence is limited to these reported samples.
Which to pick: Choose A05231 for a human colon tissue lysate precedent, or A05231-1 for a human cell lysate precedent (HeLa, HepG2, 293T, or T-47D). Both have WB images; the reported sample and conditions are the useful distinction.
Source: BosterBio LIMA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9UHB6.
- Human Protein Atlas. LIMA1 tissue expression.
- PMC12016094 — target-verified WB comparison
- PMC11662050 — target-verified WB comparison
- PMC12340046 — target-verified WB comparison