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- Table of Contents
OSCAR / Osteoclast-associated immunoglobulin-like receptor · Western blot design guide
Design a Western Blot for OSCAR
Real validated OSCAR Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-OSCAR 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
OSCAR 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 | ~30.5 kDa | |
| Observed band | ~30 kDa | |
| Gel | 10% (catalog A02961-3) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 7 isoform(s) |
Section 1
Real Curated OSCAR Western Blot Protocols
The A02961-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEL, human SIHA, rat testis (catalog A02961-3) |
| Gel % | 10% (catalog A02961-3) |
| Load | 30 ug; reducing conditions (catalog A02961-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02961-3) |
| Membrane | nitrocellulose membrane (catalog A02961-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02961-3) |
| Primary antibody | A02961-3 · 0.5 μg/mL (catalog A02961-3) |
| Primary incubation | overnight at 4°C (catalog A02961-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02961-3) |
| Secondary incubation | 1.5 hour at RT (catalog A02961-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02961-3) |
| Detection | ECL (catalog A02961-3) |
Section 2
What Is the Expected OSCAR Western Blot Band Size?
OSCAR is predicted at 30.5 kDa and observed near 30 kDa; the cause of the small difference is not established.
| Band near 30 kDa | matches the empirical OSCAR band in reducing whole-cell lysates |
| Band above 30 kDa | could reflect N-linked glycosylation at Asn48 or Asn145; a visible shift is unproven |
| Band below the precursor | could reflect cleavage of the signal peptide at residues 1–18 |
| Multiple bands | could reflect OSCAR isoforms, though distinct migration is unproven |
| Little or no band in whole-cell lysate | could reflect secretion of isoform 1 |
💡Expected OSCAR appearanceUniProt predicts 30.5 kDa, while antibody QC detects OSCAR at approximately 30 kDa in reducing whole-cell lysates; confirm band identity with a positive control and an independent antibody.
| UniProt predicted mass | 30.5 kDa for the supplied sequence; the empirical band is approximately 30 kDa |
| N-linked glycosylation at Asn48 | may increase apparent size, but no visible shift is established |
| N-linked glycosylation at Asn145 | may increase apparent size, but no visible shift is established |
| Signal peptide at residues 1–18 | cleavage may make the mature protein smaller than its precursor |
| Isoforms 1, 2, 3, 4, 5, 6, and 7 | may differ in size; their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | isoform 1 is secreted | check conditioned medium alongside a positive-control lysate |
| Band higher than expected | glycosylation at Asn48 or Asn145 is possible | compare untreated and deglycosylated samples and verify band identity |
| Band lower than expected | signal-peptide cleavage is possible | check with an independent OSCAR antibody and a positive control |
| Broad smear instead of sharp band | heterogeneous occupancy of the two N-linked glycosylation sites is possible | compare untreated and deglycosylated samples |
| Multiple bands | isoforms 1, 2, 3, 4, 5, 6, and 7 are annotated, but distinct bands are unproven | confirm candidate bands with an independent antibody or isoform-specific control |
| Weak or no signal | secreted isoform 1 may be depleted from the sampled lysate | check conditioned medium and run a positive control |
Experiment evidence
Western blot images showing OSCAR band
A02961-3 · OSCAR
A02961 · OSCAR
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for OSCAR Western blot
🧪For positive controls for OSCAR in Western blot, you can use no HPA-validated sample; the supplied HPA record lists no positive candidates.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: Isoform 1 is secreted, so conditioned medium may give a clearer signal than whole-cell lysate; HPA provides no validated tissue controls.
HPA tissue expression evidence for OSCAR
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced OSCAR Western Blot Tips
Deeper troubleshooting and optimisation questions for OSCAR, answered from its protein features.
How should OSCAR 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.
Which OSCAR isoforms might produce different bands?
Isoforms · Seven isoforms are listed. Their sequence changes include residues 13..25 in isoforms 4 and 6, residue 24 in 3 and 7, deletion of 129..134 in 5, and replacement of 219..282 in 2, 3, and 6. These are UniProt canonical coordinates. Check which isoforms an antibody can recognize before assigning bands.
Could glycosylation affect OSCAR migration?
PTM · UniProt lists N-linked glycosylation at asparagines 48 and 145, using canonical sequence coordinates. Glycosylation could affect migration, but these annotations do not predict a particular band position or prove that an additional band is glycosylated OSCAR.
Does this guide establish induction of OSCAR?
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 OSCAR?
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 A02961-3 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 I quantify OSCAR across samples?
Quantitation · Compare the same resolved band and sample fraction across samples. OSCAR has seven annotated isoforms, and isoform 1 is annotated as secreted; changes in isoform recognition or sample fraction could complicate interpretation of a single band’s intensity.
Does a ~30 kDa OSCAR band match its predicted mass?
Interpretation · The observed ~30 kDa band is close to the predicted 30.5 kDa mass, but that agreement alone does not establish identity. Signal peptide processing and glycosylation are annotated; the supplied features do not establish a visible shift or explain any specific mass difference.
UniProt annotates isoform 1 as secreted and also lists membrane-related keywords for OSCAR. Record which sample fraction was tested when comparing results. The supplied features do not specify the location of every isoform.
Consider the annotated isoform sequence changes, N-linked glycosylation at canonical positions 48 and 145, and the signal peptide at 1..18. None of these features alone identifies an unexpected band; verify antibody recognition of the relevant sequence before assigning it to OSCAR.
UniProt annotates a signal peptide at canonical residues 1..18. Its processing could change the protein’s sequence mass relative to the full-length 30.5 kDa prediction, but the supplied evidence does not establish a specific processed band size.
UniProt lists one disulfide bond without supplying its coordinates. Compare reduced and non-reduced samples if band behavior is in question, but the annotation alone does not predict the direction or size of a mobility change.
UniProt annotates isoform 1 as secreted and also lists membrane-related keywords for OSCAR. Record which sample fraction was tested when comparing results. The supplied features do not specify the location of every isoform.
Consider the annotated isoform sequence changes, N-linked glycosylation at canonical positions 48 and 145, and the signal peptide at 1..18. None of these features alone identifies an unexpected band; verify antibody recognition of the relevant sequence before assigning it to OSCAR.
UniProt annotates a signal peptide at canonical residues 1..18. Its processing could change the protein’s sequence mass relative to the full-length 30.5 kDa prediction, but the supplied evidence does not establish a specific processed band size.
UniProt lists one disulfide bond without supplying its coordinates. Compare reduced and non-reduced samples if band behavior is in question, but the annotation alone does not predict the direction or size of a mobility change.
Boster reagents
OSCAR 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-OSCAR Antibody Picoband®
Cat # A02961-3
Real WB data
Anti-OSCAR Antibody
Cat # A02961
Two the supplier anti-OSCAR antibodies have reported Western blot images. A02961-3 shows an approximately 30 kDa band in human cell and rat and mouse testis lysates; A02961 reports a blot of K562 lysate, with limited detail on the observed band.
Which to pick: For rat samples, choose A02961-3, the only listed antibody with rat reactivity and a rat testis blot. For human or mouse samples, A02961-3 has more detailed blot evidence; A02961 is listed for human and mouse but shows only a K562 blot.
Source: BosterBio OSCAR gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8IYS5.
- Human Protein Atlas. OSCAR tissue expression.
- PMC13176349 — target-verified WB comparison
- PMC11801645 — target-verified WB comparison