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- Table of Contents
OSBP / Oxysterol-binding protein 1 · Western blot design guide
Design a Western Blot for OSBP
Real validated OSBP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OSBP 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
OSBP 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 | ~89.4 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (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 | Phosphorylated + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated OSBP Western Blot Protocols
The PA1949 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Rat Kidney, Rat Spleen, Rat Lung (catalog PA1949) |
| 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 | PA1949; 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 OSBP Western Blot Band Size?
OSBP has a predicted monomer mass of 89.4 kDa; oligomerization and phosphorylation are documented, but their effects on blot migration are not demonstrated here.
| Band near 89.4 kDa | Consistent with monomeric OSBP; confirm identity with controls |
| Band near 179 kDa | Could reflect a homodimer if it survives sample preparation |
| Band near 268 kDa | Could reflect a homotrimer if it survives sample preparation |
| Weak band in a soluble fraction | Some OSBP may remain associated with Golgi or endoplasmic reticulum membranes |
💡Expected OSBP appearanceUniProt predicts an 89.4 kDa OSBP monomer, but no empirical band size is supplied; confirm a band near that position with ordinary identity controls.
| Predicted monomer mass | Places the reference band near 89.4 kDa |
| Homodimer formation | Could yield a band near twice the monomer mass if the complex persists |
| Homotrimer formation | Could yield a band near three times the monomer mass if the complex persists |
| Phosphoserines at residues 190, 193, 198, 238, 240, 338, and 345 | Modification is documented, but a visible migration change is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | OSBP may be poorly recovered from membrane-associated pools | Check soluble and membrane-enriched fractions with a positive control |
| Band higher than expected | OSBP homodimers or homotrimers may persist | Compare reducing and denaturing sample conditions |
| Band lower than expected | Possible degradation or nonspecific detection; no cleavage feature is supplied | Use fresh lysate and confirm identity with an independent antibody |
| Multiple bands | Persistent oligomers or nonspecific detection may contribute | Compare sample conditions and confirm bands with an independent antibody |
| Weak or no signal | Recovery may vary between cytosolic and membrane-associated OSBP pools | Check extraction and loading with a positive control |
| Fragments below expected size | Possible sample degradation | Process lysate promptly with protease inhibitors and compare a fresh sample |
Experiment evidence
Western blot images showing OSBP band
PA1949 · OSBP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OSBP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for OSBP in Western blot, you can use adrenal gland tissue, which has high HPA expression.
Positive control: Adrenal gland (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 HPA not-detected tissue is listed, so use OSBP knockdown or knockout for a clear negative control.
HPA tissue expression evidence for OSBP
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
| Spleen | cells in white pulp | Low | Protein (IHC) | HPA → |
| Adipose tissue | adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced OSBP Western Blot Tips
Deeper troubleshooting and optimisation questions for OSBP, answered from its protein features.
Where should the main OSBP band appear?
Band shift · OSBP has a predicted mass of 89.4 kDa. Use that as a starting point for identifying a band, not an exact migration target; no observed band position is supplied.
Do annotated isoforms explain multiple OSBP bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. Do not assign extra bands to specific isoforms from these features alone; compare their apparent sizes and antibody recognition before interpreting them.
Could phosphorylation cause a shifted OSBP band?
PTM · UniProt lists 14 phosphorylation sites, including clusters at residues 190–198 and 377–389. A phosphatase-treated aliquot can help test whether phosphorylation contributes to an apparent shift. These features alone do not establish that a shift will be visible.
Check whether an antibody targets a region containing UniProt phosphosites 190, 193, 198, 238, 240, 338, 345, 351, 377, 379, 382, 385, 386, or 389. Residue 2 is N-acetylalanine. These are UniProt coordinates; antibody or paper numbering may differ.
Check whether an antibody targets a region containing UniProt phosphosites 190, 193, 198, 238, 240, 338, 345, 351, 377, 379, 382, 385, 386, or 389. Residue 2 is N-acetylalanine. These are UniProt coordinates; antibody or paper numbering may differ.
Does this guide establish induction of OSBP?
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 OSBP?
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 PA1949 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 OSBP bands be quantified across samples?
Quantitation · Keep the sampled fraction consistent because OSBP is reported in both cytosol and membrane-associated locations. Quantify a consistently identified OSBP band; if several bands appear, establish their identity before combining their signals.
Which fractions should be checked for OSBP?
Interpretation · OSBP is reported in cytosol and the perinuclear region and as a peripheral protein at Golgi and endoplasmic reticulum membranes. If a fraction gives a weak band, check the other relevant fractions before concluding OSBP is absent.
OSBP is described as a homodimer or homotrimer and interacts with VAPA and RELCH. Those annotations make complexes worth considering, but they do not identify a higher Western blot band. Compare the band with the 89.4 kDa monomer prediction and verify its OSBP specificity.
OSBP is described as a homodimer or homotrimer and interacts with VAPA and RELCH. Those annotations make complexes worth considering, but they do not identify a higher Western blot band. Compare the band with the 89.4 kDa monomer prediction and verify its OSBP specificity.
Boster reagents
OSBP 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-OSBP1 Antibody Picoband®
Cat # PA1949
PA1949 is an anti-OSBP antibody listed for rat, mouse, and human. Its Western blot image shows rat kidney, spleen, and lung lysates and HeLa and A549 cell lysates. The supplied evidence does not show a mouse sample or independent validation.
Which to pick: PA1949 is the only listed antibody. It has a Western blot image with rat tissue and human cell lysates; mouse reactivity is listed, but no mouse sample appears in the supplied image caption.
Source: BosterBio OSBP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P22059.
- Human Protein Atlas. OSBP tissue expression.
- PMC9791146 — target-verified WB comparison
- PMC7355079 — target-verified WB comparison
- PMC9338045 — target-verified WB comparison