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SORBS1 / Sorbin and SH3 domain-containing protein 1 · IHC design guide
Design Immunohistochemistry for SORBS1
Plan SORBS1 IHC-P around general cytoplasmic staining (HPA tissue IHC). Use heart cardiomyocytes or smooth muscle cells as positive controls (HPA tissue IHC), and start the catalog antibody at 0.5–1 μg/mL (datasheet A04426-3).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SORBS1 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 | General cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in cardiomyocytes and smooth muscle cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04426-3) | |
| Positive control | Duodenum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Intensity regulation is unreported (UniProt) | |
| Isoform / epitope | 12 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended SORBS1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A04426-3) with published SORBS1 IHC methods from two breast cancer studies (PMC7922363; PMC9206459).
| Sample | Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A04426-3) |
| Fixation | Image fixative and duration unreported (datasheet A04426-3); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A04426-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04426-3) |
| Primary antibody | Rabbit anti-SORBS1, 0.5-1μg/ml (datasheet A04426-3) |
| Primary incubation | Overnight at 4 °C (datasheet A04426-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04426-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SORBS1-positive staining in glandular cells of duodenum (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 (datasheet: A04426-3); test pH 9.0 during optimization (PMC7922363; PMC9206459).
Section 2
What Is the Expected SORBS1 Staining Pattern?
SORBS1 is mainly cytoplasmic in tissue IHC, with high staining reported in cardiomyocytes, smooth muscle cells, and duodenal and small intestinal glandular cells (HPA tissue IHC). Junctional and focal adhesion localization is also plausible (UniProt Q9BX66 subcellular location). SORBS1 has no transmembrane segment (UniProt Q9BX66 topology). HPA rates tissue IHC reliability Enhanced but reports low consistency with RNA expression (HPA tissue IHC).
| Clear cytoplasmic stain in cardiomyocytes or smooth muscle cells (HPA: High in both). | This matches the reported tissue IHC pattern. Score the named cells and their intensity, while retaining HPA's low RNA–staining consistency caveat (HPA tissue IHC). |
| Strong stain confined to vessel lumens or extracellular spaces, with little cellular signal. | That distribution does not match HPA's general cytoplasmic pattern (HPA tissue IHC). Treat it as possible detection artefact and compare with a no-primary control (general IHC practice). |
| Strong stain in adipocytes or lymph node non-germinal center cells (HPA: Not detected in these cells). | Unexpected staining raises cross-reactivity or endogenous detection activity as possibilities; check control sections and the detection system before assigning SORBS1 positivity (general IHC practice). |
| A diffuse haze covers cells and surrounding tissue. | Diffuse background prevents reliable cell-specific scoring. Review blocking, antibody concentration, washing, and detection controls (general IHC practice); it does not establish SORBS1 distribution. |
| No signal in heart cardiomyocytes or smooth muscle cells (HPA: High in both). | Check a known-positive section, antibody and detection controls, and retrieval conditions before interpreting the specimen as negative (general IHC practice). HPA's High level is an observed pattern, not a guarantee for every section (HPA tissue IHC). |
💡Expected SORBS1 appearanceA convincing positive is predominantly cytoplasmic staining with High signal in cardiomyocytes, smooth muscle cells, or duodenal or small intestinal glandular cells (HPA tissue IHC); isolated luminal or diffuse stain warrants an artefact check (general IHC practice).
| Tissue and cell selection (HPA tissue IHC) | Heart cardiomyocytes and smooth muscle cells provide reported High examples; adipocytes and lymph node non-germinal center cells are reported Not detected (HPA tissue IHC). |
| Antibody evidence (HPA antibody validation) | HPA027559 and HPA043084 have Enhanced IHC ratings; CAB032850 is Approved (HPA antibody validation). These ratings do not remove the tissue profile's RNA-consistency caveat (HPA tissue IHC). |
| IF/ICC: where should signal localize? (HPA subcellular ICC-IF) | Mainly at focal adhesion sites, with plasma membrane signal also supported; centrosome localization is uncertain (HPA subcellular ICC-IF). These cell-image findings need not appear as resolvable puncta in chromogenic tissue IHC. |
| Compartment and topology (UniProt Q9BX66) | SORBS1 is associated with cytoskeleton, junctions, focal adhesions, and sometimes nucleus; it has no transmembrane segment (UniProt Q9BX66 subcellular location and topology). Do not infer a cell-surface-only IHC pattern. |
| Isoforms and epitope scope (UniProt Q9BX66) | UniProt lists 12 isoforms (UniProt Q9BX66). The supplied antibody records give no epitope positions, so isoform-specific staining cannot be predicted from them. |
| Fixation sensitivity | Target-specific fixation sensitivity is unreported in the supplied UniProt and HPA records. Optimize retrieval against controls as general IHC practice, without assuming a SORBS1-specific fixation effect. |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive section is blank. | A detection, antibody, or retrieval failure is possible (general IHC practice); cardiomyocytes and smooth muscle cells are reported High (HPA tissue IHC). | Run a known-positive section alongside a detection control; confirm antibody use and compare retrieval conditions (general IHC practice). |
| Stain is uniformly dark across the section. | Excess antibody or detection background may obscure the reported cytoplasmic pattern (general IHC practice; HPA tissue IHC). | Compare a no-primary control, review blocking and washes, and titrate the primary antibody (general IHC practice). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible, especially where HPA reports Not detected cells (HPA tissue IHC; general IHC practice). | Compare cell types on the same section and examine no-primary and detection controls before scoring them positive (general IHC practice). |
| Signal appears only in lumens or extracellular material. | The location conflicts with HPA's general cytoplasmic tissue pattern (HPA tissue IHC) and may reflect artefact. | Check tissue morphology and no-primary controls; score cellular staining separately from deposits (general IHC practice). |
| A different antibody gives a different tissue pattern. | HPA validation status varies by antibody, and HPA reports low consistency between staining and RNA expression (HPA antibody validation; HPA tissue IHC). | Record antibody ID and compare staining in the same named cell types with matched controls (general IHC practice). |
| The section shows nuclear staining without the expected cytoplasmic pattern. | UniProt includes nuclear localization, but HPA describes general cytoplasmic tissue IHC (UniProt Q9BX66 subcellular location; HPA tissue IHC). | Check morphology and controls before calling nuclear stain artefactual or SORBS1-positive; document the compartment separately (general IHC practice). |
Experiment evidence
IHC & IF images showing SORBS1 staining
A04426-3 · IHCIHC analysis of SORBS1 using anti-SORBS1 antibody (A04426-3). SORBS1 was detected in paraffin-embedded section of human rectal cancer tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000095637-SORBS1/tissue/cerebral+cortex#imid_20880503
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000095637-SORBS1/tissue/colon#imid_20880421
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000095637-SORBS1/tissue/liver#imid_20880422
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000095637-SORBS1/tissue/kidney#imid_20880415
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA027559; green = SORBS1, blue = nucleus, red = microtubules.
HPA · ICC-IF Hep-G2Human Protein Atlas immunofluorescence, Hep-G2, antibody HPA027559; green = SORBS1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SORBS1 IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain strongly (HPA: High in cardiomyocytes). Use lymph node non-germinal center cells as the negative comparison (HPA: Not detected); extracellular spaces on the heart slide should show only background chromogen, while neighboring cell types should not be assumed negative.
Positive control tissue: Duodenum (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SORBS1 in Hep-G2, SK-MEL-30, U2OS, with annotated localisation: Focal adhesion sites (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a species- and class-matched rabbit IgG isotype control (caption: rabbit primary antibody); and SORBS1 knockout material or a validated peptide-block control. In heart muscle, block endogenous peroxidase and check endogenous biotin background when using the caption’s streptavidin-biotin detection (caption: SABC with DAB).
⚠️Feasibility: The selected paraffin-section caption reports EDTA pH 8.0 heat retrieval and 1 µg/ml primary antibody overnight at 4°C; its fixative is unreported, and no target-specific fixation window or fixation effect is reported (caption: paraffin-embedded section; fixative not stated). Retrieval was used in that example, but whether SORBS1 staining depends on it is unreported; the supplied evidence does not establish that frozen sections or IF are easier. For IF/ICC interpretation, focal adhesion staining has support, while centrosome localization is uncertain (HPA: focal adhesion sites supported; centrosome uncertain).
HPA tissue IHC evidence for SORBS1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Low 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 |
|---|---|---|---|---|
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
| Small intestine | Glandular cells | High | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SORBS1 IHC Tips
Troubleshoot SORBS1 staining in paraffin sections by checking retrieval, cell type and subcellular pattern against the available tissue evidence.
What retrieval should I use when SORBS1 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A04426-3). The tissue-IHC caption reports this buffer but gives no heating time, so record your instrument's temperature and time before comparing runs (datasheet A04426-3; standard IHC practice). If signal remains weak, vary heating duration on adjacent sections while holding antibody concentration and detection constant, and inspect tissue morphology after each condition (standard IHC practice). Include a known positive compartment, such as cardiomyocytes or smooth muscle cells, to distinguish inadequate retrieval from a genuinely low-expressing specimen (HPA: High in cardiomyocytes and smooth muscle cells; standard IHC practice).
Could fixation explain weak or uneven SORBS1 staining?
The selected paraffin-tissue caption does not state a fixative, so SORBS1-specific sensitivity to fixation is unknown (datasheet A04426-3: fixative not stated). Record the fixative, fixation interval and processing history for each section; uneven exposure can change antigen accessibility and tissue preservation in routine IHC (standard IHC practice). Compare sections with matched processing before changing retrieval or antibody concentration, and check whether weak staining follows the tissue edge or damaged areas (standard IHC practice). Do not infer fixation tolerance from the reported tissue-expression pattern or from SORBS1's cellular locations and phosphorylation sites (HPA: tissue IHC profile; UniProt Q9BX66: localisation and modified residues).
Which staining compartments are plausible for SORBS1 in tissue IHC?
Expect a predominantly cytoplasmic tissue-IHC pattern, while examining cell borders and adhesion-associated regions where morphology permits (HPA: general cytoplasmic expression; HPA subcellular: focal adhesion sites and plasma membrane). SORBS1 also associates with the cytoskeleton and adherens junctions, and nuclear localisation is documented in specific contexts (UniProt Q9BX66: subcellular location). Treat diffuse nuclear DAB across unrelated cell types cautiously rather than assuming that every nuclear signal reflects those contexts (UniProt Q9BX66: nuclear localisation; standard IHC practice). Compare the stained cells with adjacent negative-control sections and expected positive cell populations before assigning a compartment-specific result (HPA: tissue IHC; standard IHC practice).
How can isoforms or epitope placement affect the SORBS1 IHC result?
SORBS1 has 12 annotated isoforms, but the selected tissue-IHC caption does not identify the antibody epitope (UniProt Q9BX66: isoforms; datasheet A04426-3: tissue-IHC caption). Therefore, a negative stain cannot establish absence of every isoform, especially if the recognised sequence is missing or inaccessible in some variants (UniProt Q9BX66: alternative splicing; standard IHC interpretation). The protein contains a SoHo domain at residues 366–469 and three SH3 domains, which can guide review of any subsequently supplied immunogen map (UniProt Q9BX66: domains). Resolve discordant samples with an independently mapped epitope or an orthogonal expression assay, using matched cell populations for comparison (standard IHC practice).
How should I check SORBS1 localisation by multiplex immunofluorescence?
Pair SORBS1 with a marker identifying the expected cell population, such as cardiomyocytes or smooth muscle cells, before interpreting colocalisation (HPA: High in cardiomyocytes and smooth muscle cells; standard IF practice). Choose spectrally separated fluorophores and inspect an unstained section so tissue autofluorescence is not mistaken for signal; a longer-wavelength channel can help when background is strong (standard IF practice). Because SORBS1 has no transmembrane segment and has cytoskeletal and focal-adhesion localisation, use permeabilisation appropriate for an intracellular epitope, with the antibody's mapped epitope checked if available (UniProt Q9BX66: topology and localisation; standard IF practice). Compare cell borders and adhesion sites with cytoplasmic signal, and include single-label controls when assessing apparent overlap (HPA subcellular: focal adhesion sites and plasma membrane; standard IF practice).
How do I reduce diffuse brown signal in SORBS1 sections?
Compare the specimen with a section processed without primary antibody to identify signal arising from secondary reagent or detection chemistry (standard IHC practice). The selected caption uses biotinylated secondary antibody, a streptavidin-biotin detection complex and DAB, so check endogenous biotin and peroxidase contributions when background follows tissue structures (datasheet A04426-3; standard IHC practice). Apply a peroxidase block and appropriate serum or protein block, then adjust washing or antibody concentration while retaining a positive-control section (standard IHC practice). The caption reports 10% goat serum blocking and 1 µg/mL primary antibody overnight at 4°C; treat these as tested caption conditions, not universal optima (datasheet A04426-3).
What should I score when comparing SORBS1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue IHC is generally cytoplasmic while focal-adhesion and membrane localisation is also supported (HPA: tissue profile; HPA subcellular: focal adhesion sites and plasma membrane). For comparable sections, record percent positive cells and an H-score using intensity grades 0–3, yielding a range of 0–300 (standard IHC practice). Normalise counts to the number of evaluable cells in the same annotated region, or report positive-cell density per mm² when cell density differs (standard IHC practice). Exclude necrosis, folds and edge artefact consistently, and report the chosen threshold alongside the score (standard IHC practice).
How can I distinguish true SORBS1 staining from artefact?
Look for reproducible staining in plausible cells, including cardiomyocytes, smooth muscle cells and intestinal glandular cells, alongside the reported general cytoplasmic pattern (HPA: tissue IHC). Interpret isolated intense staining in an unexpected cell population cautiously, particularly when a matched no-primary control shows similar colour (HPA: tissue IHC; standard IHC practice). Edge-limited signal, necrotic deposits and endogenous peroxidase colour can mimic DAB positivity, so examine tissue integrity and the detection control before scoring (standard IHC practice). Nuclear signal needs independent support in its specific context, and tissue staining should be interpreted cautiously because antibody staining and RNA expression showed low consistency (UniProt Q9BX66: nuclear localisation; HPA: reliability description).
Boster reagents
Best SORBS1 / Sorbin and SH3 domain-containing protein 1 IHC Antibodies
The IHC-validated anti-SORBS1 antibody has real staining data from a human paraffin section (catalog image caption); IF/ICC data are absent (catalog applications and images).
Real IHC data
Anti-SORBS1 ® Antibody
Cat # A04426-3
A04426-3 lists human IHC reactivity for paraffin sections (catalog applications and reactivity). Its IHC image shows SORBS1 staining in human rectal cancer tissue (catalog image caption).
Which to pick: Choose A04426-3 for human paraffin-section IHC; its caption documents EDTA retrieval at pH 8.0 and primary antibody at 1 μg/ml (catalog image caption). The fixative is unreported (catalog image caption). No IF/ICC or cross-species choice is supported by this payload: A04426-3 lists human reactivity and no IF/ICC application or image (catalog applications, reactivity, and images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9BX66 (SRBS1_HUMAN, Sorbin and SH3 domain-containing protein 1).
- Human Protein Atlas. SORBS1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SORBS1 subcellular location (ICC-IF): Mainly localized to the focal adhesion sites. In addition localized to the plasma membrane and centrosome..
- Human Protein Atlas. SORBS1 antibody validation summary (3 antibodies).
- Study on CCDC69 interfering with the prognosis of patients with breast cancer through PPAR signal pathway. European journal of histochemistry : EJH 2021 — PMC7922363.
- Development and Validation of a Prognostic Classifier Based on Lipid Metabolism-Related Genes for Breast Cancer. Journal of inflammation research 2022 — PMC9206459.
- Down-regulation and clinical significance of Sorbin and SH3 domain-containing protein 1 in bladder cancer tissues. IET systems biology 2023 — PMC10116029.
- Development and Validation of a Novel PPAR Signaling Pathway-Related Predictive Model to Predict Prognosis in Breast Cancer. Journal of immunology research 2022 — PMC9184151.
- PubMed PMID:11374898 — UniProt-cited evidence.
- PubMed PMID:11371513 — UniProt-cited evidence.
- PubMed PMID:11532984 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SORBS1 (Sorbin and SH3 domain-containing protein 1). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04426-3). Validated IHC antibody: A04426-3 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Duodenum (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human rectal cancer tissue; fixative not specified (datasheet A04426-3); 2. Fixation: Image fixative and duration unreported (datasheet A04426-3); 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 retrieval: EDTA pH 8.0 (datasheet A04426-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04426-3); 8. Primary antibody: Rabbit anti-SORBS1, 0.5-1μg/ml (datasheet A04426-3); 9. Primary incubation: Overnight at 4 °C (datasheet A04426-3). Watch for: Fixation: Keep fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 12 isoforms; epitope coverage is unspecified (UniProt); Regulation: Intensity regulation is unreported (UniProt). Evidence: UniProt Q9BX66, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the focal adhesion sites. In addition localized…, Datasheet A04426-3, PMC7922363, PMC9206459. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SORBS1/immunohistochemistry-ihc-guide.