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SDCBP2 / Syntenin-2 · IHC design guide
Design Immunohistochemistry for SDCBP2
Plan SDCBP2 chromogenic IHC in paraffin sections using gastrointestinal and gallbladder tissue staining as a reference (HPA tissue IHC). This guide covers fixation, the catalog antibody’s staining conditions, and interpretation where presumed off-target binding may complicate scoring (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SDCBP2 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 in GI tract and gallbladder (HPA tissue IHC) | |
| Staining pattern | GI enterocytes/endocrine cells and gallbladder glands: cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A13491-2) | |
| Positive control | Colon+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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding can complicate scoring (HPA tissue IHC) | |
| Regulation | Expressed in differentiated keratinocytes (UniProt) | |
| Isoform / epitope | 2 isoforms (1, 3); epitope impact undetermined (UniProt) |
Section 1
Recommended SDCBP2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A13491-2) with the published SDCBP2 staining protocol for LUAD tissue microarrays (PMC12705533).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A13491-2) |
| Fixation | Image fixative and duration unreported (datasheet A13491-2); 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 A13491-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A13491-2) |
| Primary antibody | Rabbit anti-SDCBP2, 2-5 μg/ml (datasheet A13491-2) |
| Primary incubation | Overnight at 4 °C (datasheet A13491-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A13491-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SDCBP2-positive staining in enterocytes - Microvilli of colon (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in gastrointestinal tract and gallbladder. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A13491-2); the published LUAD excerpt does not specify retrieval (PMC12705533).
Section 2
What Is the Expected SDCBP2 Staining Pattern?
In paraffin-section IHC, expect SDCBP2 staining chiefly in the cytoplasm of gastrointestinal and gallbladder cells, with strong staining in the specific cell populations listed below (HPA: tissue IHC, Enhanced). Apical microvillar staining is also plausible in colon enterocytes (HPA: High in colon enterocyte microvilli; UniProt Q9H190: apical enrichment). SDCBP2 has no transmembrane segment; nuclear localization is documented in a separate cellular context (UniProt Q9H190: topology and localization).
| Strong cytoplasmic staining in small-intestinal enterocytes or stomach and gallbladder glandular cells. | This matches the reported high-staining populations (HPA: High in each population). Assess the identified cells and their compartments; a uniformly brown section does not reproduce this selective pattern (general IHC practice). |
| Prominent staining at colon enterocyte microvilli, with nearby cells less conspicuous. | An apical pattern is credible here (HPA: High in colon enterocyte microvilli; UniProt Q9H190: apical enrichment). Record it separately from broader cytoplasmic staining; a sharp surface signal alone does not establish that every colon cell is positive (HPA: cell-specific IHC finding). |
| Staining is confined to extracellular material or unrelated structures, with no expected cellular pattern. | Treat this as a localization mismatch requiring review (HPA: chiefly cytoplasmic tissue profile; UniProt Q9H190: cytoplasm and intracellular locations). Nuclear staining alone needs context: nucleoplasm is supported by ICC-IF and is not automatically an artefact (HPA: subcellular ICC-IF). |
| Strong staining appears in adipocytes or lung alveolar cells instead of the expected positive populations. | Investigate cross-reactivity or endogenous detection activity before assigning SDCBP2 positivity (HPA: not detected in adipocytes and alveolar cells; HPA: presumed off-target binding disregarded). These HPA findings are cell-specific comparisons, not a claim that every cell in those tissues is negative. |
| The section is diffusely brown, or a known-positive gastrointestinal population has no detectable staining. | Diffuse color limits cell-specific interpretation; absent signal in a reported high-staining population leaves assay performance unresolved (HPA: high gastrointestinal populations; general IHC practice). Check controls and staining conditions before calling the tissue negative. |
💡Expected SDCBP2 appearanceCall a positive result when the expected gastrointestinal or gallbladder cells show discernible cytoplasmic staining, often strong and sometimes apical in colon enterocyte microvilli (HPA: tissue IHC, High; UniProt Q9H190: apical enrichment); broad color in HPA-not-detected cell populations is a reason to investigate a false positive, not confirm one (HPA: tissue IHC; general IHC practice).
| Cell population and tissue context | HPA reports High staining in small-intestinal enterocytes, stomach and gallbladder glandular cells, and duodenal and rectal endocrine cells; it reports other populations as Medium, Low, or Not detected (HPA: tissue IHC). Score the named cells rather than averaging the whole section (general IHC practice). |
| Antibody evidence | The tissue profile has Enhanced reliability, with medium staining-to-RNA consistency and presumed off-target binding observed and disregarded (HPA: tissue IHC). HPA064490 has Enhanced IHC validation; HPA005804 has no stated IHC status in this payload (HPA: antibody validation). Interpret unexpected staining cautiously. |
| Intracellular distribution and topology | SDCBP2 has cytoplasmic, nuclear, and membrane-associated localizations but no transmembrane segment (UniProt Q9H190: localization and topology). Tissue IHC is chiefly cytoplasmic, while ICC-IF supports nucleoplasm and cytosol and lists Golgi localization as uncertain (HPA: tissue IHC and subcellular ICC-IF). Match the compartment claim to the assay. |
| Processing and isoforms | The record lists a single 1–292 chain, no signal peptide or propeptide, and two isoforms (UniProt Q9H190: processing and isoforms). These facts do not identify the antibody epitope or establish isoform coverage; avoid explaining an absent IHC signal by cleavage or shedding without further evidence. |
| IF/ICC Q: Is nuclear SDCBP2 staining expected? | A: Mainly nucleoplasmic localization is supported in ICC-IF; cytosol is also supported, while Golgi localization is uncertain (HPA: subcellular ICC-IF). Use the separate IF/ICC guide for its workflow; these cellular findings do not prescribe a paraffin-section IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive stomach glandular cells or small-intestinal enterocytes show no signal. | A reported High population is absent, so the result cannot yet distinguish a weak assay from a sample-specific finding (HPA: tissue IHC). | Confirm that the expected cells are present and review section quality, retrieval, antibody dilution, detection, and a positive control (general IHC practice). Do not infer SDCBP2-specific retrieval or fixation sensitivity from HPA staining levels. |
| Color develops across the section and cell boundaries are hard to score. | Diffuse background can arise from detection or blocking conditions (general IHC practice); HPA also flags presumed off-target binding in its tissue assessment (HPA: reliability note). | Examine a no-primary control, review blocking and detection steps, and adjust assay conditions using an expected positive population as a reference (general IHC practice; HPA: High gastrointestinal populations). |
| Adipocytes or lung alveolar cells stain strongly. | That conflicts with the reported Not detected status for those named cells; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Compare the named cells with a positive-control section and inspect a no-primary control (general IHC practice). Report the discrepancy without extending the HPA negative call to every cell in the tissue. |
| Only extracellular deposits or a uniform surface rim stain. | The pattern does not match the chiefly cytoplasmic tissue profile or the reported intracellular and apical localizations (HPA: tissue IHC; UniProt Q9H190: localization). | Check morphology and counterstain, then compare with the expected cell-specific pattern (general IHC practice; HPA: tissue IHC). Keep colon microvillar staining distinct from a nonspecific rim (HPA: High in colon enterocyte microvilli). |
| A nuclear signal appears and is being marked as automatically false. | SDCBP2 can occupy nuclear compartments, and nucleoplasmic ICC-IF localization is supported (UniProt Q9H190: localization; HPA: subcellular ICC-IF). The tissue IHC summary, however, is chiefly cytoplasmic (HPA: tissue IHC). | Record the assay and cell type, compare tissue staining with its cytoplasmic reference pattern, and assess controls before interpreting nuclear IHC staining (HPA: tissue IHC; general IHC practice). |
| Different antibodies give different tissue patterns. | HPA reports presumed off-target binding in the tissue assessment, and the supplied IHC validation status differs between HPA064490 and HPA005804 (HPA: reliability note and antibody validation). | Prioritize the antibody with stated Enhanced IHC validation, compare staining in the same named positive and not-detected populations, and document any discordance (HPA: antibody validation and tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing SDCBP2 staining
A13491-2 · IHCIHC analysis of Syntenin 2/SDCBP2 using anti-Syntenin 2/SDCBP2 antibody (A13491-2). Syntenin 2/SDCBP2 was detected in a paraffin-embedded se
A13491-2 · IHCIHC analysis of Syntenin 2/SDCBP2 using anti-Syntenin 2/SDCBP2 antibody (A13491-2). Syntenin 2/SDCBP2 was detected in a paraffin-embedded se
A13491-2 · IFIF analysis of Syntenin 2/SDCBP2 and Tubulin beta using anti-Syntenin 2/SDCBP2 antibody (A13491-2) and anti-Tubulin beta antibody (M05613-4)
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000125775-SDCBP2/tissue/cerebral+cortex#imid_19808445
HPA · DuodenumHuman Protein Atlas IHC, Duodenum. Source: https://www.proteinatlas.org/ENSG00000125775-SDCBP2/tissue/duodenum#imid_19807570
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000125775-SDCBP2/tissue/colon#imid_19807579
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000125775-SDCBP2/tissue/liver#imid_19807572
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005804; green = SDCBP2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005804; green = SDCBP2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SDCBP2 IHC & IF
🧪Run colon first and expect staining in enterocyte microvilli (HPA: High in colon enterocyte microvilli). Use heart muscle as the negative tissue, with cardiomyocytes at background (HPA: Not detected in heart muscle cardiomyocytes); compare other cells on the colon slide with the stained microvilli as internal background, without assuming every other cell type is SDCBP2-negative.
Positive control tissue: Colon (Enterocytes - Microvilli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SDCBP2 in A-431, RT-4, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a host- and isotype-matched rabbit IgG control (selected-SKU caption: rabbit primary), and SDCBP2 knockout material or a validated peptide-block control. Quench endogenous peroxidase and check luminal material for nonspecific DAB signal in colon sections (selected-SKU caption: HRP/DAB detection; HPA: colon enterocyte microvilli).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not report a fixative. Heat retrieval in EDTA at pH 8.0 produced staining in the selected-SKU paraffin-section example, but retrieval dependence is unreported (selected-SKU caption: heat-mediated EDTA retrieval). Paraffin IHC is demonstrated by that caption; HPA ICC-IF images support cellular IF assessment, but the supplied evidence does not establish that IF or frozen sections are easier (HPA: ICC-IF images in A-431, RT-4 and U2OS).
HPA tissue IHC evidence for SDCBP2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. Presumed off target binding observed and disregarded.). 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 |
|---|---|---|---|---|
| Colon | Enterocytes - Microvilli | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | High | Protein (IHC) | HPA → |
| Small intestine | Enterocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SDCBP2 IHC Tips
Troubleshoot SDCBP2 staining in paraffin sections by checking retrieval, cell type, compartment, and controls before comparing signal intensity.
How should I retrieve SDCBP2 when paraffin sections stain weakly?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A13491-2). The catalog antibody stained a paraffin-embedded human breast cancer section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet A13491-2). Keep section thickness, heating, cooling, and antibody incubation consistent across a retrieval comparison so that a changed signal can be attributed to retrieval (standard IHC practice). Include a positive gastrointestinal reference and a no-primary control; high enterocyte or glandular staining provides a cell-specific benchmark, while the control reveals detection background (HPA tissue IHC; standard IHC practice).
Does fixation change SDCBP2 staining in paraffin sections?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state its fixative (datasheet A13491-2). Record each specimen’s fixative and fixation interval, then compare sections processed together before attributing a weak result to biological absence (standard IHC practice). Apply the documented EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody consistently when assessing specimens with different fixation histories (datasheet A13491-2; standard IHC practice). Include a positive tissue reference on each run, and evaluate morphology alongside staining so that poor preservation is recognized during interpretation (HPA tissue IHC; standard IHC practice).
Which cellular compartments should show credible SDCBP2 staining?
Assess cytoplasmic staining in the appropriate gastrointestinal or gallbladder cells first: tissue IHC reports predominantly cytoplasmic expression there (HPA tissue IHC). High reported signals include colon enterocyte microvilli, small-intestinal enterocytes, and stomach or gallbladder glandular cells (HPA tissue IHC). Nuclear signal can still be biologically plausible because SDCBP2 occupies nucleoplasm, nucleolus, and nuclear speckles, while ICC/IF chiefly supports nucleoplasm with additional cytosol (UniProt Q9H190 subcellular; HPA subcellular). Score nuclear and cytoplasmic compartments separately, and check whether an unexpected membrane or diffuse signal follows cell boundaries and controls; SDCBP2 has no transmembrane segment (UniProt Q9H190 topology; standard IHC practice).
Could isoforms or epitope access explain discordant SDCBP2 staining?
SDCBP2 has 2 listed isoforms, 1 and 3, so confirm which sequence the antibody recognizes before interpreting an absent stain (UniProt Q9H190 isoforms; standard IHC practice). The record places PDZ domains at residues 108–187 and 192–267, but the supplied product caption does not identify its epitope (UniProt Q9H190 domains; datasheet A13491-2). Without an epitope map, a compartment difference cannot be assigned to an isoform or masked domain from IHC alone (standard IHC practice). Compare staining with a separately validated antibody to a documented, distinct epitope, using the same tissue compartments and controls when assessing discordance (standard IHC practice).
How can I check SDCBP2 localisation with multiplex immunofluorescence?
Pair SDCBP2 with a marker for the expected cell population, such as an enterocyte or glandular-cell marker, and verify each marker’s specificity in the specimen (HPA tissue IHC; standard IF practice). Select fluorophores after checking tissue autofluorescence in an unstained section, and separate marker channels to limit spectral overlap (standard IF practice). Permeabilise sufficiently to expose cytosolic and nuclear epitopes, while confirming the antibody’s epitope location if available; SDCBP2 has no transmembrane segment (UniProt Q9H190 topology; standard IF practice). Compare nucleoplasmic and cytosolic signal with single-stain and no-primary controls, since those compartments have ICC/IF support (HPA subcellular; standard IF practice).
How do I reduce diffuse DAB or nonspecific SDCBP2 staining?
Begin with a no-primary control and inspect whether DAB persists in the same cells or tissue structures (standard IHC practice). The selected caption used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-linked secondary for 30 minutes at 37°C (datasheet A13491-2). Check the general peroxidase-blocking and wash steps, then adjust antibody exposure only against matched positive tissue and background controls (standard IHC practice). Treat broad staining cautiously because the HPA tissue profile explicitly notes presumed off-target binding, although its listed reliability is Enhanced (HPA tissue IHC).
How should I quantify SDCBP2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring; gastrointestinal and gallbladder tissue IHC is mainly cytoplasmic, whereas nuclear localisation is also documented (HPA tissue IHC; UniProt Q9H190 subcellular). For each defined compartment, record the percentage of positive target cells and staining intensity, then calculate an H-score if intensity categories are reproducible (standard IHC practice). Alternatively, report positive-cell density per mm² of viable tissue when cell counts or tissue area differ across sections (standard IHC practice). Normalise comparisons to the same eligible cell population, viable area, retrieval run, and detection settings, and exclude necrotic or damaged regions consistently (standard IHC practice).
What distinguishes true SDCBP2 signal from staining artefact?
Favour staining in the expected cell population and compartment, particularly cytoplasm in gastrointestinal or gallbladder cells, when the positive control performs as expected (HPA tissue IHC; standard IHC practice). Interpret nuclear signal separately because nucleoplasm has ICC/IF support and UniProt also lists nucleolar and speckle localisation (HPA subcellular; UniProt Q9H190 subcellular). Flag staining confined to section edges, necrotic regions, or the no-primary control, and investigate endogenous peroxidase before assigning those patterns to SDCBP2 (standard IHC practice). A membrane pattern alone requires independent confirmation: SDCBP2 can associate with intracellular membranes but has no transmembrane segment, and HPA notes presumed off-target tissue staining (UniProt Q9H190 subcellular and topology; HPA tissue IHC).
Boster reagents
Best SDCBP2 / Syntenin-2 IHC Antibodies
A13491-2 is listed for human IHC and IF (catalog: applications and reactivity), with images of paraffin-embedded human cancer tissue and U20S cells (catalog: IHC and IF image captions).
Real IHC data
Anti-Syntenin 2/SDCBP2 Antibody ®
Cat # A13491-2
A13491-2 will render with an IHC figure from a paraffin-embedded human breast cancer section (catalog: IHC image caption). The same SKU is listed for IF/ICC and has an IF image from U20S cells (catalog: applications and IF image caption).
Which to pick: Choose A13491-2 for human paraffin-section IHC: its caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (catalog: IHC image caption). For IF/ICC, A13491-2 is also listed and has a U20S-cell IF image at 5 μg/ml (catalog: applications and IF image caption). No cross-species choice is supported because A13491-2 lists human reactivity only (catalog: reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9H190 (SDCB2_HUMAN, Syntenin-2).
- Human Protein Atlas. SDCBP2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SDCBP2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the Golgi apparatus and cytosol..
- Human Protein Atlas. SDCBP2 antibody validation summary (2 antibodies).
- SDCBP2 promotes tumor progression and is a novel ferroptosis-related prognostic biomarker in lung adenocarcinoma. Frontiers in immunology 2025 — PMC12705533.
- SDCBP-AS1 destabilizes β-catenin by regulating ubiquitination and SUMOylation of hnRNP K to suppress gastric tumorigenicity and metastasis. Cancer communications (London, England) 2022 — PMC9648392.
- Dipeptidase 1 (DPEP1) is a marker for the transition from low-grade to high-grade intraepithelial neoplasia and an adverse prognostic factor in colorectal cancer. British journal of cancer 2013 — PMC3738143.
- PubMed PMID:11152476 — UniProt-cited evidence.
- PubMed PMID:11780052 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SDCBP2 (Syntenin-2). Expected localisation Cytoplasmic staining in GI tract and gallbladder (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A13491-2). Validated IHC antibody: A13491-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Colon (Enterocytes - Microvilli); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A13491-2); 2. Fixation: Image fixative and duration unreported (datasheet A13491-2); 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 A13491-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A13491-2); 8. Primary antibody: Rabbit anti-SDCBP2, 2-5 μg/ml (datasheet A13491-2); 9. Primary incubation: Overnight at 4 °C (datasheet A13491-2). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Presumed off-target binding can complicate scoring (HPA tissue IHC); Isoform / epitope: 2 isoforms (1, 3); epitope impact undetermined (UniProt); Regulation: Expressed in differentiated keratinocytes (UniProt). Evidence: UniProt Q9H190, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A13491-2, PMC12705533. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SDCBP2/immunohistochemistry-ihc-guide.