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- Table of Contents
SCARB2 / Lysosome membrane protein 2 · Western blot design guide
Design a Western Blot for SCARB2
Source-linked SCARB2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SCARB2 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
SCARB2 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 | ~54.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Breast (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Blocking peptide control | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked SCARB2 Western Blot Protocol Options
The A05090 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse liver tissue lysate (catalog A05090) |
| 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 | Western blot analysis of LIMP2 in mouse liver tissue lysate with LIMP2 antibody at 1 μg/mL in (A) the absence and (B) presence of blocking peptide (catalog A05090) |
| Primary antibody | A05090 · 1 μg/mL (catalog A05090) |
| 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 SCARB2 Western Blot Band Size?
SCARB2 is predicted at 54.3 kDa; N-linked glycans and isoforms could affect migration, but no empirical band size or distinct isoform pattern is supplied.
| Band near 54.3 kDa | consistent with the predicted mass, pending identity controls |
| Band above 54.3 kDa | N-linked glycosylation could affect migration; the apparent size is unverified |
| Broad band or smear | heterogeneous N-linked glycosylation is possible but unconfirmed |
| Several bands at different positions | isoforms 1 and 2 or glycoforms are possible; distinct isoform migration is unverified |
| Weak or absent band in soluble lysate | the lysosome membrane protein may be poorly recovered |
💡Expected SCARB2 appearanceSCARB2 has a predicted mass of 54.3 kDa, but no empirical band size is supplied; glycosylation may alter migration, so confirm candidate bands with the supplied blocking peptide control.
| Predicted mass | 54.3 kDa is the sequence-based reference, not a measured band |
| N-linked glycosylation at Asn45 | may increase apparent size if occupied |
| N-linked glycosylation at Asn68 | may increase apparent size if occupied |
| N-linked glycosylation at Asn105 | may increase apparent size if occupied |
| Isoforms 1 and 2 | may differ in size; their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SCARB2 is a lysosome membrane protein and may be poorly recovered in soluble lysate | Check membrane recovery and use a membrane-enriched preparation |
| Band higher than expected | N-linked glycosylation may affect migration | Compare with a deglycosylated sample and check peptide blocking |
| Band lower than expected | Reduced glycosylation or an isoform is possible, but neither is established for the band | Compare treated and untreated samples and check peptide blocking |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible | Compare with a deglycosylated sample |
| Multiple bands | Isoforms 1 and 2 or glycoforms are possible | Use peptide blocking to assess band identity and compare deglycosylated samples |
| Weak or no signal | Limited recovery of this membrane protein may reduce signal | Check membrane extraction and sample loading |
Experiment evidence
Western blot images showing SCARB2 band
A05090 · SCARB2
M05090 · SCARB2
A05090-1 · SCARB2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SCARB2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SCARB2 in Western blot, you can use adrenal gland tissue, which has high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Breast (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Breast provides a not-detected tissue control, but SCARB2’s lysosomal membrane location makes effective membrane protein extraction important.
HPA tissue expression evidence for SCARB2
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 → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Caudate | glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Breast | glandular cells | Not detected | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SCARB2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SCARB2, answered from its protein features.
How should SCARB2 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 SCARB2 isoforms affect band interpretation?
Isoforms · Isoform 2 lacks canonical residues 93–235. Compare any distinct bands with this deletion, but do not assign bands by size alone. Coordinates here refer to the supplied canonical sequence; positions downstream of the deletion differ in isoform 2.
The canonical sequence lists N-linked sites at positions 45, 68, 105, 206, 224, 249, 304, 325, 412, and 430. Isoform 2 lacks canonical residues 93–235, including sites 105, 206, and 224. These are canonical UniProt coordinates, not renumbered isoform 2 positions.
The canonical sequence lists N-linked sites at positions 45, 68, 105, 206, 224, 249, 304, 325, 412, and 430. Isoform 2 lacks canonical residues 93–235, including sites 105, 206, and 224. These are canonical UniProt coordinates, not renumbered isoform 2 positions.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of SCARB2?
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.
What transfer method to use for SCARB2 Western blot?
Transfer · SCARB2 is annotated as a multipass lysosome membrane protein. Check transfer efficiency for SCARB2 and retain membrane-associated material during sample preparation. The supplied features do not establish a specific transfer method or settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05090 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 SCARB2 bands be quantified?
Quantitation · Use the same SCARB2 band definition across samples and account for its two isoforms if they resolve separately. Because SCARB2 is a multipass lysosome membrane protein, keep membrane recovery consistent. The supplied features do not identify a reference band or validate combining distinct bands.
Why might SCARB2 migrate differently from its predicted 54.3 kDa mass?
Interpretation · SCARB2 has ten annotated N-linked glycosylation sites, so its apparent mass may differ from the predicted 54.3 kDa. No observed band mass is supplied, and site annotations alone cannot establish the size or cause of a shift.
SCARB2 has two annotated disulfide bonds. If bands differ between reducing and nonreducing preparations, compare them under matched conditions. The supplied features do not establish that the bonds produce a distinct Western blot band.
Compare their migration with the 54.3 kDa predicted mass while considering the ten annotated N-linked sites and isoform 2’s deletion of canonical residues 93–235. Record whether samples were reduced. These features suggest checks, but none alone identifies an observed band.
SCARB2 has two annotated disulfide bonds. If bands differ between reducing and nonreducing preparations, compare them under matched conditions. The supplied features do not establish that the bonds produce a distinct Western blot band.
Compare their migration with the 54.3 kDa predicted mass while considering the ten annotated N-linked sites and isoform 2’s deletion of canonical residues 93–235. Record whether samples were reduced. These features suggest checks, but none alone identifies an observed band.
Boster reagents
SCARB2 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-LIMP2 SCARB2 Antibody
Cat # A05090
Real WB data
Anti-LIMPII Rabbit Monoclonal Antibody
Cat # M05090
Real WB data
Anti-Scavenging Receptor SRB2/SCARB2 Antibody Picoband®
Cat # A05090-1
Three the supplier anti-SCARB2 antibodies list human, mouse, and rat reactivity and have Western blot images. A05090 shows a mouse liver blocking-peptide comparison; M05090 shows HeLa and mouse liver; A05090-1 shows human, rat, and mouse lysates. No publication evidence is supplied.
Which to pick: Choose A05090-1 for the widest range of illustrated samples, noting that its reported ~80 kDa band differs from the expected 54 kDa. Consider M05090 for HeLa or mouse liver, or A05090 for its mouse liver blocking-peptide comparison.
Source: BosterBio SCARB2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q14108.
- Human Protein Atlas. SCARB2 tissue expression.
- PMC3260287 — target-verified WB comparison
- PMC10901359 — target-verified WB comparison
- PMC3765843 — target-verified WB comparison