This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
SH3RF2 / E3 ubiquitin-protein ligase SH3RF2 · IHC design guide
Design Immunohistochemistry for SH3RF2
Plan chromogenic SH3RF2 IHC-P with a catalog antibody starting at 10 μg/mL (datasheet: A11489). Compare staining with high Purkinje cell signal and undetected adipocyte signal, while accounting for uncertain staining–RNA consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SH3RF2 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 tissue staining (HPA tissue IHC); nuclear location (UniProt) | |
| Staining pattern | Cytoplasmic in several tissues; high in Purkinje cells (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11489) | |
| Caveat | Low staining–RNA consistency limits interpretation (HPA tissue IHC) | |
| Regulation | Higher in colon cancer than normal colon (UniProt) | |
| Isoform / epitope | Three isoforms; check epitope coverage (UniProt) |
Section 1
Recommended SH3RF2 IHC & IF Protocols
The catalog antibody's IHC-P protocol is accompanied by 2 published SH3RF2 chromogenic IHC protocols (PMC12269186; PMC10776562).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A11489); 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 Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear 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-SH3RF2, 10 μg/mL (datasheet A11489) |
| 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 | SH3RF2-positive staining in purkinje cells of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min (page retrieval: nuclear antigen).
Section 2
What Is the Expected SH3RF2 Staining Pattern?
SH3RF2 is annotated in the nucleus (UniProt Q8TEC5) and has no transmembrane segment (UniProt Q8TEC5 topology). In tissue IHC, HPA instead reports cytoplasmic staining in several tissues, strongest in cerebellar Purkinje cells and moderate in selected glandular and kidney tubular cells (HPA tissue IHC). Treat that slide pattern as a reference with limited confidence: HPA rates its tissue IHC reliability Uncertain because staining and RNA expression have low consistency (HPA tissue IHC).
| Purkinje cells show clear cytoplasmic chromogen above nearby background. | This matches HPA's High Purkinje cell staining and its reported cytoplasmic tissue pattern (HPA tissue IHC). Assess signal within identified cells against a negative control (general IHC practice). A match supports interpretation of the staining pattern, but HPA's Uncertain reliability prevents treating it alone as definitive target validation (HPA tissue IHC). |
| Signal is confined to cell borders, extracellular material, or an unrelated tissue structure. | Those locations do not match the reported cytoplasmic tissue IHC pattern (HPA tissue IHC) or the nuclear annotation (UniProt Q8TEC5). Treat the distribution as a possible artefact and check whether it persists in controls (general IHC practice). Nuclear staining requires separate assessment because HPA reports nucleoplasm in ICC-IF (HPA subcellular). |
| Strong staining appears in alveolar cells or adipocytes while the reference-positive cells are unstained. | HPA reports alveolar cells and adipocytes as Not detected, versus High staining in Purkinje cells (HPA tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; inspect the no-primary control and cell morphology (general IHC practice). A negative HPA observation is a comparison point, not proof that every specimen must be negative (HPA tissue IHC: Uncertain). |
| Pale chromogen covers many cell types and the surrounding section without a defined cellular pattern. | Diffuse background makes the reported cell-specific pattern hard to score (HPA tissue IHC; general IHC practice). Compare with a no-primary control, then review blocking, washing, antibody concentration, and chromogen development (general IHC practice). Avoid calling uniformly weak color SH3RF2-positive solely because it appears in a tissue listed as positive (HPA tissue IHC: Uncertain). |
| No stain is visible in Purkinje cells on the test section. | Purkinje cells are HPA's strongest listed tissue IHC example (High; HPA tissue IHC). Check that the cells are present and that the detection run and tissue controls worked (general IHC practice). Because HPA rates this IHC profile Uncertain, one blank section does not establish absence of SH3RF2 protein (HPA tissue IHC). |
💡Expected SH3RF2 appearanceA plausible chromogenic IHC positive is distinct cytoplasmic signal in Purkinje cells (High) or selected glandular and kidney tubular cells (Medium); broad extracellular or cell-border staining is suspect (HPA tissue IHC; UniProt Q8TEC5 topology).
| Tissue pattern confidence | HPA labels the tissue IHC profile Uncertain because antibody staining and RNA expression have low consistency (HPA tissue IHC). Use its cell-level examples to plan comparisons, then require convincing morphology and controls before interpreting a specimen (general IHC practice). |
| Compartment evidence | UniProt annotates nucleus, and HPA ICC-IF reports enhanced nucleoplasmic localisation; HPA tissue IHC describes cytoplasmic expression (UniProt Q8TEC5; HPA subcellular; HPA tissue IHC). Record nuclear and cytoplasmic staining separately. The supplied sources do not resolve that assay-dependent difference. |
| Isoforms and antibody recognition | UniProt lists 3 SH3RF2 isoforms (UniProt Q8TEC5). No epitope location or isoform recognition data are supplied, so an isoform-specific staining prediction is unsupported. Interpret an unexpected negative result with that limitation rather than assigning it to a particular isoform. |
| Protein topology and processing | UniProt lists no transmembrane segment, signal peptide, or propeptide and describes the chain as residues 1–729 (UniProt Q8TEC5). A crisp membrane-only or extracellular pattern therefore needs scrutiny. These annotations do not establish how paraffin processing affects antibody binding. |
| IF/ICC Q&A: should its signal occupy the IHC compartment? | HPA reports enhanced nucleoplasmic localisation in ICC-IF, while tissue IHC is described as cytoplasmic and Uncertain (HPA subcellular; HPA tissue IHC). Score each application against its own reported pattern; a nuclear IF image does not by itself validate a cytoplasmic IHC signal. |
| Situation | Likely cause | Next action |
|---|---|---|
| Purkinje cells remain blank although the section contains identifiable Purkinje cells. | The IHC run may have insufficient sensitivity, or the HPA reference pattern may not reproduce in this specimen (general IHC practice; HPA tissue IHC: Uncertain). | Check a working positive control and detection reagents, then review the antibody's IHC-P instructions for retrieval and dilution; no target-specific retrieval condition or dilution is supplied here (general IHC practice). |
| The whole section shows brown haze that obscures cellular boundaries. | Excess antibody or detection background can reduce contrast (general IHC practice). HPA's cell-specific observations cannot be evaluated through diffuse color (HPA tissue IHC). | Compare a no-primary control; review blocking, washes, antibody concentration, and chromogen development, then score only resolved cellular signal (general IHC practice). |
| Alveolar cells or adipocytes stain strongly. | Those cell types are reported Not detected by HPA, so cross-reactivity or endogenous detection activity is possible (HPA tissue IHC; general IHC practice). | Check no-primary and detection controls, inspect cell identity, and review endogenous enzyme blocking for chromogenic detection (general IHC practice). Keep HPA's Uncertain reliability in view (HPA tissue IHC). |
| Only membranes or extracellular regions show crisp signal. | This conflicts with the cytoplasmic tissue IHC description and the lack of a transmembrane segment (HPA tissue IHC; UniProt Q8TEC5 topology). | Recheck morphology and control staining, then compare the distribution with the reported positive cell types before scoring it as SH3RF2 (general IHC practice; HPA tissue IHC). |
| Nuclear signal is visible, but the expected cytoplasmic tissue pattern is weak. | The sources differ by application: UniProt places SH3RF2 in the nucleus, HPA ICC-IF reports nucleoplasm, and HPA tissue IHC reports cytoplasm (UniProt Q8TEC5; HPA subcellular; HPA tissue IHC). | Document both compartments and examine controls. Do not automatically discard nuclear staining or count it as confirmation of the uncertain tissue IHC pattern (HPA tissue IHC: Uncertain; general IHC practice). |
| Moderate signal in kidney tubules varies between areas of the section. | HPA reports Medium staining in tubular cells but provides no rule that every tubule must stain equally (HPA tissue IHC). Section quality or detection variation can complicate scoring (general IHC practice). | Score identified tubular cells and local background separately, inspect tissue preservation, and compare with controls before calling an area positive or negative (general IHC practice). |
Experiment evidence
IHC & IF images showing SH3RF2 staining
A11489 · IHCImmunohistochemistry of SH3RF2 in human brain tissue with SH3RF2 antibody at 10 μg/mL.
A11489 · IFImmunofluorescence of SH3RF2 in human brain tissue with SH3RF2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000156463-SH3RF2/tissue/cerebral+cortex#imid_18057655
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000156463-SH3RF2/tissue/colon#imid_18057581
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000156463-SH3RF2/tissue/liver#imid_18057590
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000156463-SH3RF2/tissue/kidney#imid_18057583
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035903; green = SH3RF2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035903; green = SH3RF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SH3RF2 IHC & IF
🧪Run cerebellum first and score Purkinje cells, where SH3RF2 staining is High (HPA: High in Purkinje cells). Use adipose tissue as a negative comparator, where adipocytes are Not detected (HPA: Not detected in adipocytes); on the cerebellum slide, neighboring cells without specific staining should remain at background, without assuming they are SH3RF2-negative.
Positive control tissue: Cerebellum (Purkinje cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SH3RF2 in A-431, U-251MG, CACO-2, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an irrelevant isotype control matched to the primary antibody’s host species and immunoglobulin class (standard IHC practice). A SH3RF2 knockout sample can provide a biological negative control; quench endogenous peroxidase for chromogenic detection, block endogenous biotin if using biotin-based detection, and assess tissue autofluorescence if performing IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A11489 brain IHC caption does not state a fixative (selected A11489 caption). Retrieval dependency is also unreported, so optimize antigen retrieval empirically for paraffin sections (standard IHC practice). The supplied evidence does not establish that frozen sections or IF are easier; in cerebellum, distinguish specific Purkinje-cell staining from background and, for IF, tissue autofluorescence (HPA: High in Purkinje cells; standard IHC/IF practice).
HPA tissue IHC evidence for SH3RF2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 |
|---|---|---|---|---|
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SH3RF2 IHC Tips
Troubleshooting SH3RF2 staining in paraffin sections, with one Q&A on translating the readout to IF/ICC.
How should I retrieve SH3RF2 before chromogenic IHC?
Start with Tris-EDTA pH 9.0 heat-induced retrieval at 95–98 °C for 20 min (page retrieval specification). Let sections cool in the retrieval buffer, then keep the antibody concentration and detection conditions constant while comparing retrieved and untreated sections (standard IHC practice). If staining is weak, optimise heating or test an alternative retrieval buffer as a documented fallback, using the same tissue and a no-primary control (standard IHC practice). The selected A11489 image reports SH3RF2 staining in human brain tissue at 10 µg/mL, but its caption gives neither retrieval conditions nor fixative, so it cannot validate this retrieval step (A11489 tissue-IHC caption).
Could fixation explain weak or uneven SH3RF2 staining?
Target-specific SH3RF2 sensitivity to fixation is unknown from the supplied evidence; the selected A11489 tissue-IHC caption does not state a fixative (A11489 tissue-IHC caption). For paraffin IHC, compare sections with documented fixation histories under the same retrieval, antibody and detection conditions before attributing a difference to fixation (standard IHC practice). Review preservation, section thickness and tissue-edge staining, and include a no-primary control to expose background unrelated to the primary antibody (standard IHC practice). Do not infer a fixation preference from reported SH3RF2 localisation, phosphorylation or tissue-staining patterns; those records do not test fixation effects (UniProt Q8TEC5; HPA tissue IHC).
Should SH3RF2 staining appear in nuclei or cytoplasm?
Assess nuclear and cytoplasmic signal separately: UniProt lists SH3RF2 in the nucleus, and HPA reports enhanced nucleoplasmic localisation in ICC/IF images (UniProt Q8TEC5; HPA subcellular). HPA tissue IHC instead describes cytoplasmic expression across several tissues, with uncertain reliability because staining and RNA data have low consistency (HPA tissue IHC). In paraffin sections, use a nuclear counterstain to define compartments and record which cell types carry each pattern (standard IHC practice). Treat a cytoplasmic-only or nuclear-only pattern as a finding requiring orthogonal confirmation, rather than forcing agreement between assays that used different specimens and readouts (HPA tissue IHC; HPA subcellular).
How can isoforms and epitope position affect SH3RF2 IHC?
Check the antibody’s documented immunogen or epitope against all 3 SH3RF2 isoforms before treating a negative section as absence of the protein (UniProt Q8TEC5). The record lists 729 residues for the canonical chain, SH3 domains at residues 125–184, 187–252 and 380–441, and a phosphoserine at 649, but supplies no epitope for A11489 (UniProt Q8TEC5; A11489 tissue-IHC caption). Compare staining with a second antibody whose epitope is documented and distinct, ideally on adjacent sections processed together (standard IHC validation practice). Interpret differences cautiously until isoform recognition and epitope accessibility have been established for the antibodies and tissue preparation used (standard IHC validation practice).
How should I adapt an SH3RF2 IHC finding for multiplex IF?
Use IF as a separate validation experiment: HPA reports enhanced nucleoplasmic SH3RF2 localisation in ICC/IF images, whereas its tissue IHC profile describes cytoplasmic staining (HPA subcellular; HPA tissue IHC). Pair SH3RF2 with a validated marker for the cell type being assessed, such as Purkinje cells in cerebellum, and inspect channels separately before interpreting overlap (HPA tissue IHC: High in Purkinje cells; standard IF practice). Choose fluorophores after checking tissue autofluorescence in an unstained section and use single-colour controls for bleed-through (standard IF practice). Because SH3RF2 has no transmembrane segment and nucleoplasmic signal is reported, permeabilise cells to allow access to intracellular epitopes, then optimise that step for the antibody and preparation (UniProt Q8TEC5 topology; HPA subcellular; standard IF practice).
What should I check when SH3RF2 chromogenic staining is diffuse?
Run a no-primary section alongside the stained section to identify detection-system colour, endogenous peroxidase activity and nonspecific tissue background (standard chromogenic IHC practice). Block endogenous peroxidase before the detection step, optimise protein blocking and washing, and shorten DAB development if colour accumulates outside cell boundaries (standard chromogenic IHC practice). Titrate the primary antibody around a documented starting point; the selected A11489 human-brain image reports 10 µg/mL, without stating that this concentration is optimal for other specimens (A11489 tissue-IHC caption). Score diffuse cytoplasmic colour cautiously because HPA reports cytoplasmic tissue staining but assigns its SH3RF2 IHC profile uncertain reliability (HPA tissue IHC).
How should I quantify SH3RF2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell type and compartment before scoring, since UniProt lists nuclear SH3RF2 while HPA tissue IHC describes cytoplasmic expression with uncertain reliability (UniProt Q8TEC5; HPA tissue IHC). Record the percentage of positive target cells and staining intensity, then calculate an H-score if intensity categories are reproducible across batches (standard IHC scoring practice). For spatial comparisons, report positive-cell density per mm² of viable, annotated tissue and normalise cell-based measurements to the number of eligible cells examined (standard IHC quantification practice). Keep exposure, DAB development, section thickness and thresholds consistent, and analyse nuclear and cytoplasmic scores separately rather than pooling them (standard IHC quantification practice).
How can I distinguish credible SH3RF2 signal from artefact?
A credible result should be cellular, reproducible across comparable sections and consistent with the scored compartment and cell population, while no-primary sections reveal detection-related colour (standard IHC validation practice). Purkinje cells in cerebellum have reported high SH3RF2 staining, but HPA rates the overall tissue-IHC profile uncertain because staining and RNA expression have low consistency (HPA tissue IHC). Investigate staining confined to section edges, necrotic areas or regions with endogenous enzyme activity before calling those cells positive (standard chromogenic IHC practice). Nuclear signal has support from UniProt and HPA subcellular data, whereas HPA tissue IHC reports cytoplasmic expression; confirm any decisive compartment-specific claim with independent evidence (UniProt Q8TEC5; HPA subcellular; HPA tissue IHC).
Boster reagents
Best SH3RF2 / E3 ubiquitin-protein ligase SH3RF2 IHC Antibodies
The IHC-validated antibody has human brain IHC and IF images (catalog image captions); its listed reactivity covers human, mouse and rat (catalog: A11489).
Real IHC data
Anti-SH3RF2 Antibody
Cat # A11489
A11489 will render with its own human brain IHC image at 10 μg/mL (catalog IHC image caption). Its human brain IF image uses 20 μg/mL (catalog IF image caption).
Which to pick: Choose A11489 for tissue IHC: its own image shows staining in human brain tissue at 10 μg/mL (catalog IHC image caption); the fixative is unreported (catalog IHC image caption). Choose A11489 for IF; its own image shows human brain tissue staining at 20 μg/mL (catalog IF image caption), while ICC is unreported (catalog applications). For work across species, A11489 lists human, mouse and rat reactivity; A11489-1 lists human and rat and has no supplied IHC image (catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TEC5 (SH3R2_HUMAN, E3 ubiquitin-protein ligase SH3RF2).
- Human Protein Atlas. SH3RF2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. SH3RF2 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. SH3RF2 antibody validation summary (2 antibodies).
- The role of SH3RF2 in lung squamous cell carcinoma and M2 polarization: insights into LZTS2 ubiquitination. Biology direct 2025 — PMC12269186.
- SH3RF2 contributes to cisplatin resistance in ovarian cancer cells by promoting RBPMS degradation. Communications biology 2024 — PMC10776562.
- PubMed PMID:19945436 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15372022 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SH3RF2 (E3 ubiquitin-protein ligase SH3RF2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC); nuclear location (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A11489 at 10 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Purkinje cells); negative Adipose tissue. 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 A11489); 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 Tris-EDTA buffer, pH 9.0 , 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-SH3RF2, 10 μg/mL (datasheet A11489); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A11489); Caveat: Low staining–RNA consistency limits interpretation (HPA tissue IHC); Isoform / epitope: Three isoforms; check epitope coverage (UniProt); Regulation: Higher in colon cancer than normal colon (UniProt). Evidence: UniProt Q8TEC5, HPA tissue IHC · Uncertain, HPA ICC-IF · nucleoplasm., Datasheet A11489, PMC12269186, PMC10776562. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SH3RF2/immunohistochemistry-ihc-guide.