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SCARB2 / Lysosome membrane protein 2 · IHC design guide
Design Immunohistochemistry for SCARB2
Plan chromogenic SCARB2 IHC on paraffin sections using the IHC-validated antibody at 2 μg/mL (datasheet: A05090-1 IHC-P). Use high staining in colon glandular cells as a positive reference (HPA tissue IHC), then assess cytoplasmic staining intensity and distribution (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SCARB2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining (HPA tissue IHC); lysosome membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Breast+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A05090); verify before use. | |
| Caveat | Staining–RNA consistency is medium (HPA tissue IHC) | |
| Regulation | Intensity regulation is unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope conservation and position are unknown (UniProt) |
Section 1
Recommended SCARB2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol accompanies this published SCARB2 protocol for tree shrew tissue sections (PMC12241082).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A05090); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SCARB2, 2.5 μg/mL (datasheet A05090) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SCARB2-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0, 95–98 °C for 20 min (page antigen retrieval); the published protocol used citric acid pH 9.0 (PMC12241082).
Section 2
What Is the Expected SCARB2 Staining Pattern?
SCARB2 is a lysosomal membrane protein with two transmembrane segments and a large lumenal region (UniProt Q14108 topology). In paraffin-section IHC, expect cytoplasmic staining in cells shown as positive by HPA, including glandular, respiratory epithelial, and glial cells (HPA tissue IHC). HPA describes the tissue profile as ubiquitous cytoplasmic expression, with Supported reliability and medium consistency between antibody staining and RNA data (HPA tissue IHC).
| Cytoplasmic staining in colon or duodenum glandular cells, bronchus respiratory epithelial cells, or cerebral cortex glial cells. | These cell and tissue combinations are reported as High by HPA (HPA tissue IHC). Record intensity in the named cell population rather than assigning one score to the whole section. A cytoplasmic result fits HPA's tissue profile; the stain alone does not identify individual lysosomes (HPA tissue IHC; UniProt Q14108 subcellular location). |
| Predominantly nuclear staining, or a sharp cell-surface rim without a cytoplasmic component. | That distribution does not fit HPA's reported cytoplasmic tissue pattern or UniProt's lysosomal membrane assignment (HPA tissue IHC; UniProt Q14108 subcellular location). Treat it as an interpretation concern; check the staining run and antibody validation before calling it SCARB2. |
| Strong signal in breast glandular cells or vaginal squamous epithelial cells. | HPA reports SCARB2 as Not detected in those particular cell populations (HPA tissue IHC). Unexpected staining warrants a check for cross-reactivity or endogenous detection activity (general IHC practice). It does not, by itself, disprove HPA's broader description of ubiquitous cytoplasmic expression (HPA tissue IHC). |
| Diffuse color across cells, extracellular areas, and the section background, obscuring cell boundaries. | A widespread deposit cannot be scored confidently as the cell-associated cytoplasmic pattern reported by HPA (HPA tissue IHC). Review background controls, blocking, wash steps, and chromogen development as general IHC checks; do not assign the diffuse deposit to SCARB2 without a distinct cellular pattern. |
| No staining in the expected positive cells of colon, duodenum, or bronchus. | HPA reports High staining in the specified glandular or respiratory epithelial cells (HPA tissue IHC). A blank result calls for run-level checks of the antibody, detection reagents, and section handling (general IHC practice). It is not enough evidence to conclude that the specimen lacks SCARB2. |
💡Expected SCARB2 appearanceCall a result positive when the expected cells show clear cytoplasmic staining, potentially High in HPA-listed positive populations; nuclear-only signal or diffuse background is suspect (HPA tissue IHC; UniProt Q14108 subcellular location).
| Tissue and cell population | HPA reports High staining in selected glandular, respiratory epithelial, glial, and cerebellar molecular-layer cells, but Not detected in the listed breast glandular and vaginal squamous epithelial cells (HPA tissue IHC). Select and score controls by the specified cell population, since tissue-wide labels can hide differences between cell types. |
| Location and membrane topology | UniProt places SCARB2 in the lysosomal membrane and annotates transmembrane segments at residues 5–27 and 434–459, with a lumenal region at 28–433 (UniProt Q14108 topology). HPA's paraffin-section readout is cytoplasmic; organelle identity cannot be established from chromogenic localization alone (HPA tissue IHC). |
| Antibody evidence | HPA lists IHC as Supported for HPA018014 and CAB015415, while its tissue profile has Supported reliability with medium staining–RNA consistency (HPA antibodies; HPA tissue IHC). These assessments support using the reported pattern as a reference, but an unexpected result still needs controls and specimen-specific interpretation. |
| Glycosylation and isoforms | UniProt annotates 10 glycosylation sites and 2 isoforms (UniProt Q14108). The supplied record does not identify the catalog antibody's epitope or show an isoform-specific staining difference. Do not infer that glycosylation, retrieval, or isoform choice explains a particular IHC intensity from these annotations alone. |
| IF/ICC Q&A: should IF show the IHC pattern? | HPA calls cytosol the approved ICC-IF location and lists A-431 images; its tissue IHC profile is cytoplasmic (HPA subcellular; HPA tissue IHC). UniProt assigns SCARB2 to the lysosomal membrane (UniProt Q14108 subcellular location). Interpret each assay in its own context; this IHC section does not establish an IF protocol or lysosome-level colocalization. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known positive cells are blank. | A failed staining or detection run is possible; the specified colon, duodenum, and bronchus cell populations are High in HPA (HPA tissue IHC). | Check a positive control section, antibody preparation, detection reagents, and development conditions (general IHC practice). Record which expected cell population was actually present before calling the tissue negative. |
| The whole section has weak, diffuse color. | Background deposition or incomplete washing can obscure cell-associated staining (general IHC practice). | Review the no-primary control, blocking and wash steps, and chromogen development (general IHC practice). Score SCARB2 only where a distinct cytoplasmic pattern is visible in an interpretable cell population (HPA tissue IHC). |
| Signal is mainly nuclear or outlines the plasma membrane. | The location conflicts with the reported cytoplasmic tissue pattern and lysosomal membrane assignment (HPA tissue IHC; UniProt Q14108 subcellular location). | Review section morphology and detection controls, then compare with a positive reference section (general IHC practice). Do not count the discordant compartment alone as confirmed SCARB2. |
| Unexpected strong staining appears in an HPA-listed negative cell population. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); HPA reports breast glandular and vaginal squamous epithelial cells as Not detected (HPA tissue IHC). | Check a no-primary control and compare staining with the surrounding cell populations (general IHC practice). Report the discrepancy by tissue and cell type rather than treating the whole organ as uniformly positive. |
| Positive and negative areas coexist in one section. | Cell populations can differ: HPA assigns levels to named cells within tissues, and reports low tissue RNA specificity overall (HPA tissue IHC). | Score the relevant cell types separately and document whether the signal is cytoplasmic (HPA tissue IHC). Avoid using a single section-wide intensity to explain a cell-specific discrepancy. |
| An IF/ICC image seems more diffuse than the IHC reference. | HPA reports an approved cytosol location for ICC-IF and a cytoplasmic tissue IHC profile; UniProt identifies the protein as lysosomal membrane-associated (HPA subcellular; HPA tissue IHC; UniProt Q14108 subcellular location). | Keep the assay readouts separate. Use the IHC cell and compartment pattern to interpret this paraffin-section result; evaluate IF/ICC localization in its own guide and controls. |
Experiment evidence
IHC & IF images showing SCARB2 staining
A05090 · IHCImmunohistochemistry of LIMP2 in human skeletal muscle tissue with LIMP2 antibody at 2.5 μg/mL.
A05090 · IFImmunofluorescence of LIMP2 in Human Skeletal Muscle tissue with LIMP2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000138760-SCARB2/tissue/cerebral+cortex#imid_3908764
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000138760-SCARB2/tissue/colon#imid_3908674
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000138760-SCARB2/tissue/liver#imid_3908675
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000138760-SCARB2/tissue/kidney#imid_3908692
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018014; green = SCARB2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018014; green = SCARB2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SCARB2 IHC & IF
🧪Run adrenal gland first; its glandular cells should stain (HPA: High in adrenal gland glandular cells). Run breast glandular cells as the negative comparator (HPA: Not detected in breast glandular cells), and assess adjacent nonglandular cells on the adrenal slide for background without assuming they are SCARB2-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Breast (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SCARB2 in A-431, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, an isotype control matched to the primary antibody’s host species, immunoglobulin class and concentration, and SCARB2-knockout tissue if available. In adrenal gland, check endogenous peroxidase background with the no-primary slide and block endogenous biotin if using avidin–biotin detection.
⚠️Feasibility: A target-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval empirically for paraffin sections. The selected A05090 skeletal-muscle IHC caption does not report a fixative (catalog caption: fixative not stated). The evidence does not establish whether frozen sections or IF are easier; distinguish adrenal tissue pigment from chromogenic signal using the no-primary control.
HPA tissue IHC evidence for SCARB2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| 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 → |
Section 3
Advanced SCARB2 IHC Tips
Troubleshoot SCARB2 staining in paraffin sections by checking retrieval, compartment, cellular context and controls before comparing signal intensity.
How should I retrieve SCARB2 when paraffin sections stain weakly?
Use citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 minutes (page retrieval setting). Cool sections in buffer, then compare a retrieval-treated section with an untreated section under identical antibody and detection conditions (standard IHC practice). If staining remains weak, test a longer citrate retrieval on a separate section while monitoring tissue damage and background (standard IHC practice). SCARB2 has a large lumenal region between two transmembrane segments, so epitope accessibility depends on the antibody’s binding site (UniProt Q14108 topology). Do not interpret stronger, widespread staining alone as improved specificity; assess compartment and controls together (standard IHC practice).
Could fixation explain inconsistent SCARB2 staining across paraffin sections?
Target-specific fixation sensitivity is unknown because the supplied SCARB2 tissue-IHC caption does not state a fixative (A05090 tissue-IHC caption). Record each specimen’s fixative and fixation duration, then compare sections processed and stained together to limit handling differences (standard IHC practice). If staining varies, assess tissue preservation and run matched sections through the same citrate pH 6.0 retrieval at 95–98 °C for 20 minutes (page retrieval setting; standard IHC practice). The published caption reports staining in human skeletal muscle at 2.5 µg/mL, but supplies no fixation comparison (A05090 tissue-IHC caption). Avoid attributing a weak result specifically to fixation without matched processing evidence (standard IHC practice).
Should SCARB2 appear punctate or diffuse in chromogenic IHC?
SCARB2 is annotated as a lysosome membrane protein with a lumenal region spanning residues 28–433 (UniProt Q14108 localisation and topology). Accordingly, assess whether cytoplasmic chromogen includes a granular or punctate component while retaining recognizable cell borders and morphology (UniProt Q14108 localisation; standard IHC practice). The tissue-IHC profile describes ubiquitous cytoplasmic expression, whereas the approved subcellular image annotation says cytosol (HPA tissue IHC; HPA subcellular). These annotations support a broad cytoplasmic description but do not establish that every brown granule is a lysosome (HPA tissue IHC; standard IHC practice). Compare the pattern with a matched negative control before assigning compartment-specific meaning (standard IHC practice).
Can the antibody distinguish SCARB2 isoforms or lumenal epitopes in sections?
SCARB2 has two annotated isoforms, but the supplied evidence does not map this antibody’s epitope or establish isoform selectivity (UniProt Q14108 isoforms; supplied antibody evidence). Its lumenal region spans residues 28–433, with short cytoplasmic regions at 1–4 and 460–478 (UniProt Q14108 topology). Ten glycosylation sites are annotated within the lumenal region, but their effect on this antibody’s staining is untested here (UniProt Q14108 glycosylation; supplied antibody evidence). Treat changes after citrate pH 6.0 retrieval as accessibility changes until an epitope-mapped reagent or orthogonal assay supports a narrower explanation (page retrieval setting; standard IHC practice). Report staining as SCARB2 immunoreactivity rather than a specific isoform (standard IHC practice).
How can IF help investigate a questionable SCARB2 IHC pattern?
Use IF as a separate assay to examine whether SCARB2 signal overlaps a marker of the stained cell type, such as cytokeratin in glandular epithelium (HPA tissue IHC; standard IF practice). Choose a fluorophore in a spectral channel with low measured tissue autofluorescence, and include single-stain controls before judging overlap (standard IF practice). SCARB2’s central region is lumenal, while both termini are cytoplasmic, so choose permeabilisation according to a known epitope’s membrane-facing side (UniProt Q14108 topology; standard IF practice). If the epitope is unmapped, compare suitable permeabilisation conditions rather than assuming access from IHC staining (supplied antibody evidence; standard IF practice). Keep IF conclusions distinct from chromogenic paraffin-section scores (standard assay interpretation practice).
What should I check when SCARB2 chromogen covers most of the section?
First compare a no-primary control and inspect whether color follows tissue edges, folds or damaged areas rather than intact cells (standard IHC practice). Apply a peroxidase block before chromogenic detection and use appropriate protein blocking and washes to limit enzyme-driven and nonspecific signal (standard IHC practice). Titrate the primary antibody against the documented 2.5 µg/mL skeletal-muscle image condition, without treating that caption as a universal working concentration (A05090 tissue-IHC caption; standard IHC practice). SCARB2’s reported tissue staining is broadly cytoplasmic, so widespread color alone cannot establish specificity (HPA tissue IHC). Retain morphology and negative controls when deciding whether the background obscures interpretable cellular staining (standard IHC practice).
How should I score SCARB2 staining across tissue regions? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because the supplied tissue-IHC profile reports cytoplasmic staining across many tissues (HPA tissue IHC; standard IHC practice). For chromogenic sections, record the percentage of positive cells and intensity grades, then calculate an H-score on the conventional 0–300 scale if intensity is reproducibly separable (standard IHC practice). Alternatively, count positive cells per mm² of viable tissue when cell density varies across regions (standard IHC practice). Normalize comparisons to the same cell type or viable tissue area, with matched staining and image thresholds (standard IHC practice). Report background exclusion and scoring rules alongside each summary (standard IHC practice).
Which SCARB2 staining patterns warrant skepticism despite a positive control?
Interpret staining in the context of SCARB2’s lysosome membrane annotation and the reported broad cytoplasmic tissue profile (UniProt Q14108 localisation; HPA tissue IHC). Strong nuclear-only color, tissue-edge accentuation, necrotic debris or color persisting without primary antibody warrants investigation before a positive call (standard IHC practice). Check cell identity against the reference pattern: glandular cells in colon are reported high, while skeletal-muscle myocytes are reported low (HPA tissue IHC). Endogenous enzyme activity can mimic chromogenic signal, so compare peroxidase-blocked sections and no-primary controls (standard IHC practice). The tissue-IHC profile has “Supported” reliability with medium staining–RNA consistency; treat unexpected differences as findings to verify, not definitive biology (HPA tissue IHC).
Boster reagents
Best SCARB2 / Lysosome membrane protein 2 IHC Antibodies
SCARB2 IHC and IF images show human skeletal muscle (A05090 image captions); the antibody lists human, mouse and rat reactivity (catalog: A05090 reactivity).
Real IHC data
Anti-LIMP2 SCARB2 Antibody
Cat # A05090
A05090 is the sole SKU that will render, with IHC-P and IF listed as applications (catalog: A05090 applications). Its images show human skeletal muscle IHC at 2.5 μg/mL and IF at 20 μg/mL (A05090 image captions).
Which to pick: For tissue IHC, choose A05090 based on its human skeletal muscle IHC image (A05090 IHC caption); the caption does not report section processing or fixative (A05090 IHC caption). For IF, choose A05090 based on its human skeletal muscle IF image (A05090 IF caption); ICC validation is unreported (catalog: A05090 applications). For cross-species studies, A05090 lists human, mouse and rat reactivity (catalog: A05090 reactivity), while the supplied IHC and IF images document human tissue only (A05090 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14108 (SCRB2_HUMAN, Lysosome membrane protein 2).
- Human Protein Atlas. SCARB2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SCARB2 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. SCARB2 antibody validation summary (2 antibodies).
- EV-A71 invades the central nervous system and affects the blood-brain barrier in a tree shrew model. Frontiers in immunology 2025 — PMC12241082.
- Delivery of human EV71 receptors by adeno-associated virus increases EV71 infection-induced local inflammation in adult mice. BioMed research international 2014 — PMC4163470.
- SCARB2 drives hepatocellular carcinoma tumor initiating cells via enhanced MYC transcriptional activity. Nature communications 2023 — PMC10517016.
- Human SCARB2 Acts as a Cellular Associator for Helping Coxsackieviruses A10 Infection. Viruses 2023 — PMC10144829.
- PubMed PMID:1374238 — UniProt-cited evidence.
- PubMed PMID:7539776 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SCARB2 (Lysosome membrane protein 2). Expected localisation Cytoplasmic staining (HPA tissue IHC); lysosome membrane (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A05090 at 2.5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Breast. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A05090); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SCARB2, 2.5 μg/mL (datasheet A05090); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific). Selected-image fixative and duration unreported (datasheet A05090); verify before use.; Caveat: Staining–RNA consistency is medium (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope conservation and position are unknown (UniProt); Regulation: Intensity regulation is unreported (UniProt). Evidence: UniProt Q14108, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A05090, PMC12241082. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SCARB2/immunohistochemistry-ihc-guide.