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- Table of Contents
OLAH / S-acyl fatty acid synthase thioesterase, medium chain · Western blot design guide
Design a Western Blot for OLAH
Source-linked OLAH Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OLAH 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
OLAH 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 | ~29.9 kDa | |
| Observed band | ~36 kDa | |
| Gel | 5–20% (catalog A05069-3) | |
| Positive control | Testis (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Acetylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked OLAH Western Blot Protocol Options
The A05069-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U20S (catalog A05069-3) |
| Gel % | 5–20% (catalog A05069-3) |
| Load | 30 ug; reducing conditions (catalog A05069-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A05069-3) |
| Membrane | nitrocellulose membrane (catalog A05069-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A05069-3) |
| Primary antibody | A05069-3 · 0.5 μg/mL (catalog A05069-3) |
| Primary incubation | overnight at 4°C (catalog A05069-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A05069-3) |
| Secondary incubation | 1.5 hour at RT (catalog A05069-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A05069-3) |
| Detection | ECL (catalog A05069-3) |
Section 2
What Is the Expected OLAH Western Blot Band Size?
OLAH is predicted at 29.9 kDa and observed at ~36 kDa on an antibody QC blot; the cause of the difference is not established.
| Band at ~36 kDa | Empirical OLAH band in a reducing whole-cell lysate blot; the reason it runs above 29.9 kDa is not established |
| Band near 29.9 kDa | Near the predicted OLAH mass; confirm identity with controls |
| Additional bands at different positions | Isoforms 1 and 2 are annotated, but distinct migration is unverified |
| Band in whole-cell lysate | Consistent with OLAH's cytosolic location |
💡Expected OLAH appearanceOLAH has a predicted mass of 29.9 kDa, while an antibody QC blot reports ~36 kDa; the cause of the difference is unestablished, so confirm band identity with suitable controls.
| Predicted OLAH mass | 29.9 kDa is the sequence-based reference; the empirical band is ~36 kDa |
| Alternative splicing | Could affect size, but distinct migration has not been shown |
| Isoform 1 | Its size relative to isoform 2 is not supplied |
| Isoform 2 | Its size relative to isoform 1 is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytosolic OLAH may be below the assay's detection limit | Check loading, antibody performance, and a known positive lysate |
| Band higher than expected | The reported ~36 kDa band exceeds the 29.9 kDa prediction for an unestablished reason | Compare with the reported blot and verify identity with an OLAH-specific control |
| Band lower than expected | A smaller isoform or protein breakdown is possible but unverified | Check sample handling and confirm the band with an OLAH-specific control |
| Multiple bands | Annotated isoforms or nonspecific antibody binding are possible | Compare antibody controls and verify each band before assigning an isoform |
| Weak or no signal | OLAH abundance or assay sensitivity may be insufficient | Check loading, transfer, and a known positive lysate |
Experiment evidence
Western blot images showing OLAH band
A05069-3 · OLAH
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OLAH Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for OLAH in Western blot, you can use testis lysate, where HPA reports high expression.
Positive control: Testis (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: Cytosolic location supports testing tissue lysates, and HPA identifies a not-detected tissue for a negative control.
HPA tissue expression evidence for OLAH
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 |
|---|---|---|---|---|
| Testis | round or early spermatids | 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced OLAH Western Blot Tips
Deeper troubleshooting and optimisation questions for OLAH, answered from its protein features.
How should OLAH 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 the two OLAH isoforms affect band interpretation?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the valine at UniProt position 55 is replaced by a longer sequence. This could change its calculated mass and antibody recognition. Interpret bands with the antibody’s target sequence and isoform coverage in mind.
That depends on its binding sequence. Isoform 2 has an alternative sequence at UniProt position 55; an antibody targeting that region may recognize the isoforms differently. Check the antibody’s epitope before labeling separate bands as specific isoforms.
That depends on its binding sequence. Isoform 2 has an alternative sequence at UniProt position 55; an antibody targeting that region may recognize the isoforms differently. Check the antibody’s epitope before labeling separate bands as specific isoforms.
Does OLAH acetylation explain a visible band shift?
PTM · UniProt lists N-acetylmethionine at position 1, using UniProt sequence numbering. That modification alone does not demonstrate a visible Western-blot shift or explain the approximately 36 kDa band. Check the numbering convention before comparing this site with antibody or paper descriptions.
Does this guide establish induction of OLAH?
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 OLAH?
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 A05069-3 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 OLAH bands be quantified across samples?
Quantitation · Use the same defined band for comparisons and establish which isoform the antibody recognizes. UniProt lists two isoforms, so combining bands could mix distinct protein forms. The approximately 36 kDa observation alone does not identify an isoform.
Why might OLAH appear near 36 kDa instead of 29.9 kDa?
Interpretation · The supplied observed band is approximately 36 kDa, while the predicted mass is 29.9 kDa. UniProt lists two isoforms and an alternative sequence in isoform 2, but these features alone do not establish the cause of the difference. Check which isoform the antibody detects before assigning the band.
Compare each band with the 29.9 kDa predicted mass, the approximately 36 kDa observed band, and the antibody’s isoform coverage. UniProt lists an alternative sequence in isoform 2 and N-acetylmethionine at position 1, but neither feature alone assigns an unexpected band. Verify band identity before interpreting it as OLAH.
Compare each band with the 29.9 kDa predicted mass, the approximately 36 kDa observed band, and the antibody’s isoform coverage. UniProt lists an alternative sequence in isoform 2 and N-acetylmethionine at position 1, but neither feature alone assigns an unexpected band. Verify band identity before interpreting it as OLAH.
Boster reagents
OLAH 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-OLAH Antibody Picoband®
Cat # A05069-3
The catalog reports one anti-OLAH antibody, A05069-3, for human reactivity. Its Western blot image uses human U20S whole-cell lysate and shows a band near 36 kDa, versus an expected 30 kDa. No additional validation evidence is supplied.
Which to pick: A05069-3 is the only listed option. Its WB image provides a starting point for human U20S lysate under the reported conditions; consider the observed 36 kDa band when interpreting results.
Source: BosterBio OLAH gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NV23.
- Human Protein Atlas. OLAH tissue expression.
- PMC9495588 — target-verified WB comparison