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UBASH3B / Ubiquitin-associated and SH3 domain-containing protein B · IHC design guide
Design Immunohistochemistry for UBASH3B
Plan paraffin-section UBASH3B IHC around the reported cytoplasmic staining, highest in lymphoid tissue (HPA tissue IHC). Use matched fixation and tissue controls to assess staining, bearing in mind the reported low consistency between staining and RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
UBASH3B 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; nucleus also annotated (HPA tissue IHC; UniProt) | |
| Staining pattern | Cytoplasmic staining, highest in lymphoid tissue (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A09570-1) | |
| Positive control | Adrenal gland+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 sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No annotated isoforms or processing; chain spans 1–649 (UniProt) |
Section 1
Recommended UBASH3B IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet A09570-1) with a published prostate cancer tissue protocol (PMC6974685).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A09570-1) |
| Fixation | Image fixative and duration unreported (datasheet A09570-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A09570-1); 20 min, 95–100 °C (standard) |
| 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-UBASH3B, 1:50-1:200 (datasheet A09570-1) |
| 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 | UBASH3B-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues with highest expression levels in lymphoid tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6.0 for the catalog antibody (datasheet A09570-1). The published protocol uses boiling sodium citrate (PMC6974685).
Section 2
What Is the Expected UBASH3B Staining Pattern?
UBASH3B is a cytoplasmic protein with a reported nuclear location and no transmembrane segment (UniProt Q8TF42). In tissue IHC, expect cytoplasmic staining in selected cells, including lymph node germinal center cells and lung macrophages (HPA: High in both). Treat the pattern as a guide rather than a definitive identity test: HPA rates its tissue IHC Approved but reports low consistency with RNA expression and pending external verification (HPA: tissue IHC reliability).
| Cytoplasmic chromogen in lymph node germinal center cells or appendix lymphoid tissue. | This matches reported high staining in those cell populations (HPA: High in lymph node germinal center cells and appendix lymphoid tissue). Judge the signal within the named cells; the HPA reliability caveat still applies (HPA: tissue IHC reliability). |
| Predominantly surface rim or extracellular staining, with little cytoplasmic signal. | Investigate the compartment assignment: UBASH3B has no transmembrane segment and is annotated in cytoplasm and nucleus (UniProt Q8TF42). This pattern alone cannot establish an artefact; compare cell morphology and controls (general IHC practice). |
| Strong staining in adipocytes or soft tissue fibroblasts while expected positive cells are also stained. | Those cell types are reported as not detected (HPA: adipocytes and soft tissue fibroblasts). Consider cross-reactivity or endogenous detection activity, then assess a reagent control and tissue morphology (general IHC practice). |
| Diffuse chromogen across cells, stroma and blank spaces, with unclear cellular boundaries. | This is difficult to score as cellular UBASH3B staining (general IHC practice). Check a no-primary control for detection background and review blocking, washes and chromogen development as general IHC steps (general IHC practice). |
| No signal in lymph node germinal center cells or lung macrophages. | These are reported high-staining populations (HPA: lymph node germinal center cells and lung macrophages). Confirm the expected cells are present, then review the antibody's IHC validation and routine assay controls before interpreting the sample as negative (general IHC practice). |
💡Expected UBASH3B appearanceCall a positive result when distinct cytoplasmic staining appears in expected cells, such as germinal center cells or macrophages, at the reported high level (HPA: tissue IHC); widespread extracellular or surface-only color warrants investigation (UniProt Q8TF42 topology; general IHC practice).
| How strong is the tissue-pattern evidence? | HPA labels the tissue IHC profile Approved, yet notes low consistency between staining and RNA expression and says external verification is pending (HPA: tissue IHC reliability). Use its high and not-detected calls as comparators, with that limitation stated. |
| Which cells provide useful comparisons? | Lymph node germinal center cells, lung macrophages and pancreatic exocrine glandular cells are reported High (HPA: tissue IHC). Adipocytes and smooth muscle cells are reported Not detected (HPA: tissue IHC). Compare named cells rather than an entire tissue. |
| What compartment should guide scoring? | HPA describes cytoplasmic expression across several tissues (HPA: tissue IHC profile). UniProt also lists the nucleus (UniProt Q8TF42). Score the cytoplasmic pattern first; interpret isolated nuclear IHC cautiously in light of the sample and controls (general IHC practice). |
| Does IF/ICC add a localisation check? | Yes: HPA reports approved cytosolic localisation with additional nucleoplasm and nuclear body localisation in ICC-IF (HPA: subcellular). This supports possible nuclear signal, but an ICC-IF image does not establish the intensity or distribution expected in a paraffin tissue section (general IHC practice). |
| What does antibody validation establish? | Two listed antibodies have Approved IHC status, HPA038605 and HPA038607; HPA038606 has Approved ICC status (HPA: antibody validation). These are application-specific labels. The supplied record does not report an Enhanced IHC designation (HPA: antibody validation). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive lymphoid cells are unstained. | The expected population may be absent from the section, or the assay may have failed; HPA reports high staining in lymph node germinal center cells (HPA: tissue IHC; general IHC practice). | Check the section's cell morphology and an appropriate positive control, then review the IHC-validated antibody and detection run (general IHC practice). |
| All compartments show a faint, even brown haze. | Nonspecific reagent or detection background can obscure cell boundaries (general IHC practice). HPA's cell-specific levels do not explain uniform haze (HPA: tissue IHC). | Inspect a no-primary control; review blocking, washing and chromogen development using general IHC workflow checks (general IHC practice). |
| Macrophages stain, but nearby adipocytes also stain strongly. | HPA reports High in lung macrophages and Not detected in adipocytes, so the adipocyte signal conflicts with its reference pattern (HPA: tissue IHC). | Verify cell identification and examine a no-primary control; if adipocyte staining persists, investigate antibody specificity (general IHC practice). |
| Only nuclei stain in an otherwise suitable positive tissue. | A nuclear location is possible (UniProt Q8TF42; HPA: ICC-IF), but tissue IHC is described chiefly as cytoplasmic (HPA: tissue IHC profile). | Check nuclear boundaries and controls, then report the nuclear-only pattern separately rather than scoring it as the expected tissue IHC pattern (general IHC practice). |
| A surface rim is the dominant staining pattern. | The rim differs from the reported cytoplasmic tissue pattern and from the absence of a transmembrane segment (HPA: tissue IHC profile; UniProt Q8TF42 topology). | Review morphology and background controls; interpret the rim cautiously unless an independent specificity check supports it (general IHC practice). |
| Different tissues give inconsistent positives and negatives. | Expression varies by named cell population, and HPA reports low staining-to-RNA consistency with external verification pending (HPA: tissue IHC). | Record cell-level localisation and intensity alongside positive and negative comparators; avoid treating disagreement with one tissue as definitive assay failure (general IHC practice). |
Experiment evidence
IHC & IF images showing UBASH3B staining
A09570-1 · IHCImmunohistochemistry of paraffin-embedded human lung cancer using UBASH3B Rabbit pAb at dilution of 1:200 .Perform high pressure antigen ret
A09570-1 · IHCImmunohistochemistry of paraffin-embedded mouse kidney using UBASH3B Rabbit pAb at dilution of 1:200 .Perform high pressure antigen retrieva
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000154127-UBASH3B/tissue/cerebral+cortex#imid_9997556
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000154127-UBASH3B/tissue/colon#imid_9997482
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000154127-UBASH3B/tissue/liver#imid_9997475
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000154127-UBASH3B/tissue/kidney#imid_9997484
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA038606; green = UBASH3B, blue = nucleus, red = microtubules.
HPA · ICC-IF HELHuman Protein Atlas immunofluorescence, HEL, antibody HPA038606; green = UBASH3B, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for UBASH3B IHC & IF
🧪Run appendix first: its lymphoid tissue is rated High for UBASH3B (HPA: appendix, lymphoid tissue). Use adipose tissue as the biological negative (HPA: adipocytes, Not detected); on the appendix slide, compare staining in lymphoid tissue with adjacent non-lymphoid cells, but the supplied HPA rows do not establish a specific internal negative cell type.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show UBASH3B in HEL, REH, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) section, a matched rabbit polyclonal isotype control (A09570-1 caption: rabbit pAb), and UBASH3B knockout material as a specificity control. Quench endogenous peroxidase in appendix sections before chromogenic detection, since leukocytes can produce misleading signal (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A09570-1 paraffin-section caption does not state a fixative. Start paraffin IHC with high-pressure retrieval in 10 mM citrate buffer at pH 6.0 and antibody dilution 1:200 (A09570-1 caption); retrieval dependence and any advantage of frozen sections are unestablished. For the separate IF/ICC workflow, HPA images in HEL, REH and U2OS show mainly cytosolic localisation, with additional nucleoplasmic and nuclear-body signal (HPA: subcellular localisation); appendix leukocytes may add endogenous-peroxidase background in chromogenic IHC (standard IHC practice).
HPA tissue IHC evidence for UBASH3B
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Endometrium | Cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Fibroblasts | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced UBASH3B IHC Tips
Troubleshooting chromogenic IHC for UBASH3B in paraffin sections, with one IF/ICC question for comparing localization.
Which retrieval conditions should I start with for UBASH3B in paraffin sections?
Start with heat-mediated retrieval in 10 mM citrate buffer, pH 6.0, using the high-pressure approach documented for the catalog antibody in paraffin-embedded human lung cancer (datasheet A09570-1). Keep section thickness, heating, cooling, and antibody incubation consistent across a pilot run so that changes in staining can be attributed to retrieval (standard IHC practice). The pictured staining used a 1:200 primary dilution, which provides a documented starting point alongside that retrieval method (datasheet A09570-1). If staining remains weak, compare a longer heating interval on adjacent sections while checking tissue integrity and background, then retain the mildest condition that gives interpretable cytoplasmic signal (standard IHC practice; HPA tissue profile).
Could fixation explain weak or uneven UBASH3B staining?
Target-specific fixation sensitivity is unknown: the catalog image identifies paraffin-embedded human lung cancer but does not state its fixative (datasheet A09570-1). Record the fixative, time in fixative, processing schedule, and section age for each specimen before attributing weak staining to UBASH3B biology (standard IHC practice). Compare adjacent sections processed together, beginning with the documented 10 mM citrate, pH 6.0 retrieval and 1:200 antibody dilution (datasheet A09570-1). If a specimen differs from a concurrently stained control, review processing records and morphology before changing retrieval or antibody concentration (standard IHC practice). Neither the reported tissue staining nor the protein's domains establish a UBASH3B-specific fixation effect (HPA tissue profile; UniProt Q8TF42).
What staining pattern is plausible for UBASH3B in tissue sections?
Prioritize cytoplasmic staining: tissue IHC reports cytoplasmic expression, while subcellular IF places UBASH3B mainly in the cytosol (HPA tissue profile; HPA subcellular). Nuclear staining is plausible because nucleoplasm and nuclear bodies are additional approved IF locations, and UniProt also lists the nucleus (HPA subcellular; UniProt Q8TF42). UBASH3B has no transmembrane segment, so a sharp membrane-only rim warrants scrutiny even though the protein interacts with receptor-associated complexes (UniProt Q8TF42). Inspect matched hematoxylin morphology and score cytoplasmic and nuclear signals separately; compare each compartment with a concurrently stained positive tissue region and a no-primary control (standard IHC practice; HPA tissue profile).
How should I troubleshoot possible epitope masking or isoform ambiguity?
The supplied record lists one chain spanning residues 1–649 and zero annotated isoforms, so these data do not support assigning different tissue patterns to named UBASH3B isoforms (UniProt Q8TF42). Its UBA domain spans residues 27–76, its SH3 domain spans 254–319, and phosphorylation is reported at residues 20 and 23 (UniProt Q8TF42). The catalog caption does not identify the antibody epitope, so none of those features establishes whether retrieval exposes or masks its binding site (datasheet A09570-1). If staining varies between similarly processed sections, first hold retrieval at pH 6.0 and compare serial-section staining before considering epitope-sensitive interpretations (datasheet A09570-1; standard IHC practice).
How can IF help assess an ambiguous chromogenic UBASH3B pattern?
Use IF/ICC as a separate localization check: reported staining is mainly cytosolic, with additional nucleoplasmic and nuclear-body signal in imaged cell lines (HPA subcellular). For tissue multiplexing, pair UBASH3B with a validated marker for the cell population under study; lung macrophages or lymph-node germinal-center cells are documented high-staining examples (HPA tissue profile; standard IF practice). Choose a fluorophore in a channel with low measured tissue autofluorescence, and include single-stain and no-primary controls before interpreting colocalization (standard IF practice). Because UBASH3B lacks a transmembrane segment, use controlled permeabilisation to access intracellular epitopes and check whether the chosen antibody tolerates those conditions (UniProt Q8TF42; standard IF practice).
How do I separate UBASH3B staining from chromogenic background?
Inspect the no-primary control for nonspecific detection-system color, and apply a peroxidase block before DAB development when using an HRP-based workflow (standard IHC practice). Run the documented 1:200 dilution beside a more dilute trial if diffuse color obscures cell boundaries; the catalog image supports 1:200 only for its stated conditions (datasheet A09570-1; standard IHC practice). Assess pigment, precipitate, folds, and section edges against the hematoxylin image rather than counting every brown deposit as positive (standard IHC practice). The reported cytoplasmic pattern and high staining in lung macrophages can guide cell-level review, but the HPA tissue dataset has low staining-to-RNA consistency (HPA tissue profile and reliability description).
What should I score when comparing UBASH3B IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because reported tissue expression is cytoplasmic and additional nuclear localization is supported by subcellular IF (HPA tissue profile; HPA subcellular). For cytoplasmic staining, record percent positive cells and an H-score from 0–300 using intensity categories 0–3; report nuclear staining separately (standard IHC scoring practice). If the question concerns infiltrating cells, count positive cells per mm² within a stated tissue compartment and normalize to the measured area or eligible-cell count (standard IHC scoring practice). Apply identical thresholds, retrieval, imaging, and exclusion rules to all sections, and note that HPA reports low consistency between staining and RNA expression (standard IHC practice; HPA reliability description).
When is a positive UBASH3B IHC result credible?
A credible result shows reproducible cell-associated staining in an expected compartment, chiefly cytoplasm, with any nuclear component evaluated separately (HPA tissue profile; HPA subcellular; standard IHC practice). Check cell identity against morphology: high staining is reported in lung macrophages and lymph-node germinal-center cells, whereas adipocytes and smooth muscle cells were not detected in the cited tissue survey (HPA tissue profile). Treat isolated edge color, necrotic deposits, and signal reproduced in a no-primary control as potential artefacts; review endogenous peroxidase when using HRP and DAB (standard IHC practice). A membrane-only pattern needs additional validation because UBASH3B has no transmembrane segment, and tissue-level conclusions should reflect the reported low consistency with RNA expression (UniProt Q8TF42; HPA reliability description).
Boster reagents
Best UBASH3B / Ubiquitin-associated and SH3 domain-containing protein B IHC Antibodies
Anti-UBASH3B antibodies cover human, mouse and rat samples (catalog: reactivity). Paraffin-section IHC images are available for all three species; IF/ICC is listed without an image (A09570-1: image captions and applications).
Real IHC data
Anti-UBASH3B Antibody
Cat # A09570-1
A09570-1 has paraffin-section IHC images from human lung cancer, mouse kidney and rat lung, and is listed for IF/ICC (A09570-1: image captions and applications). A09570-3 is listed for IHC in human, mouse and rat, but has no IHC image in the payload (A09570-3: applications, reactivity and image captions).
Which to pick: For tissue IHC, choose A09570-1 when an illustrated paraffin-section example matters: its captions show staining in all three listed species (A09570-1: IHC image captions). For IF/ICC, choose A09570-1 because IF/ICC is listed for it, whereas A09570-3 lists IHC but no IF/ICC; both are rabbit polyclonals with human, mouse and rat reactivity (catalog: applications, dilution_raw and reactivity). The A09570-1 captions specify paraffin sections and citrate retrieval, but do not report the fixative (A09570-1: IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8TF42 (UBS3B_HUMAN, Ubiquitin-associated and SH3 domain-containing protein B).
- Human Protein Atlas. UBASH3B tissue IHC expression (reliability: Approved).
- Human Protein Atlas. UBASH3B subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the nucleoplasm and nuclear bodies..
- Human Protein Atlas. UBASH3B antibody validation summary (3 antibodies).
- UBASH3B Is a Novel Prognostic Biomarker and Correlated With Immune Infiltrates in Prostate Cancer. Frontiers in oncology 2019 — PMC6974685.
- The role of disulfidptosis-driven tumor microenvironment remodeling in pancreatic cancer progression. Frontiers in immunology 2026 — PMC13008902.
- UBASH3B-mediated MRPL12 Y60 dephosphorylation inhibits LUAD development by driving mitochondrial metabolism reprogramming. Journal of experimental & clinical cancer research : CR 2024 — PMC11441236.
- PubMed PMID:11853319 — UniProt-cited evidence.
- PubMed PMID:16303743 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for UBASH3B (Ubiquitin-associated and SH3 domain-containing protein B). Expected localisation Cytoplasmic tissue staining; nucleus also annotated (HPA tissue IHC; UniProt). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A09570-1). Validated IHC antibody: A09570-1 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A09570-1); 2. Fixation: Image fixative and duration unreported (datasheet A09570-1); 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: Citrate pH 6.0 (datasheet A09570-1); 20 min, 95–100 °C (standard); 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-UBASH3B, 1:50-1:200 (datasheet A09570-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms or processing; chain spans 1–649 (UniProt); Regulation: Expression regulation not annotated (UniProt). Evidence: UniProt Q8TF42, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A09570-1, PMC6974685. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/UBASH3B/immunohistochemistry-ihc-guide.