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BUD31 / Protein BUD31 homolog · IHC design guide
Design Immunohistochemistry for BUD31
Plan chromogenic IHC-P for BUD31 with the catalog antibody at 1:50–1:200 (datasheet: 1:50–1:200). Assess nuclear and cytoplasmic staining using colon glandular cells, reported at medium intensity, as a tissue reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BUD31 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 | Nuclear and cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Glandular and other cells show nuclear and cytoplasmic signal (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Adrenal gland+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A11662) | |
| Caveat | Cytoplasmic staining may accompany the expected nuclear signal (HPA tissue IHC; UniProt) | |
| Regulation | Staining intensity regulation is not established (UniProt) | |
| Isoform / epitope | 2 isoforms; antibody epitope coverage is unreported (UniProt; datasheet) |
Section 1
Recommended BUD31 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with 3 published BUD31 tissue IHC methods (PMC10094631; PMC9587234; PMC10214398).
| Sample | Paraffin-embedded human tonsil carcinoma tissue; fixative not specified (datasheet A11662) |
| Fixation | Image fixative and duration unreported (datasheet A11662); 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-BUD31, 1:50-1:200 (datasheet A11662) |
| 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 | BUD31-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic and nuclear expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min (page antigen retrieval); citrate pH 6.0 is a published alternative (PMC10094631).
Section 2
What Is the Expected BUD31 Staining Pattern?
BUD31 is a nuclear protein with no transmembrane segment (UniProt P41223). In tissue IHC, HPA reports cytoplasmic and nuclear staining in several tissues, including medium staining in colon glandular cells and bronchial respiratory epithelial cells (HPA: tissue IHC). The HPA tissue profile is Approved, with medium consistency between staining and RNA data (HPA: tissue IHC). Interpret a new slide against those observed cell and tissue patterns, allowing for variable signal.
| Nuclear staining, with or without cytoplasmic staining, in the expected cells. | This fits UniProt's nuclear localization and HPA's observed cytoplasmic and nuclear tissue pattern (UniProt P41223; HPA: tissue IHC). Judge the result by the stained cells and their compartments together; cytoplasmic staining alone is less persuasive for a protein whose supported IF location is nucleoplasm (HPA: subcellular ICC-IF). |
| Strong staining confined to membranes or extracellular material, with no convincing cellular pattern. | Treat this as a suspect compartment pattern. BUD31 has no transmembrane segment or signal peptide, and its annotated location is the nucleus (UniProt P41223). Check section morphology and controls before calling it BUD31; the tissue IHC profile also permits cytoplasmic staining (HPA: tissue IHC). |
| Strong staining in a cell population recorded as not detected by HPA. | For example, HPA reports no detection in adrenal gland glandular cells or spleen red-pulp cells (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous detection activity, then compare a positive tissue and detection-only control. A discrepancy is a prompt to investigate, not proof of either cause. |
| A diffuse haze covers cells and tissue spaces, obscuring nuclei. | Read this as background until a cell-resolved pattern emerges. Check the no-primary or detection-only control for nonspecific detection activity, then optimize blocking, washing and detection conditions (general IHC practice). HPA's cytoplasmic and nuclear description refers to identifiable cells, not uniform staining across the section (HPA: tissue IHC). |
| No signal in colon glandular cells or bronchial respiratory epithelial cells. | Both populations have medium staining in the supplied HPA profile (HPA: tissue IHC). Confirm tissue preservation and a working detection control; review retrieval and antibody conditions using the antibody's IHC instructions (general IHC practice). One negative slide does not overturn an Approved profile with only medium staining–RNA consistency (HPA: tissue IHC). |
💡Expected BUD31 appearanceCall a result positive when identifiable cells show nuclear staining, possibly with cytoplasmic staining, at a clearly discernible intensity in an HPA medium-staining population; isolated membrane staining or section-wide haze is suspect (UniProt P41223; HPA: tissue IHC; general IHC practice).
| Compartment evidence | UniProt places BUD31 in the nucleus; HPA tissue IHC describes cytoplasmic and nuclear expression (UniProt P41223; HPA: tissue IHC). Keep both observations in view when scoring chromogenic sections rather than requiring every positive cell to look exclusively nuclear. |
| Choice of comparison tissue | HPA records medium staining in colon glandular cells and bronchial respiratory epithelial cells, low staining in prostate glandular cells, and no detection in adrenal gland glandular cells (HPA: tissue IHC). These categories guide comparisons; they are not guarantees for an individual section. |
| Antibody evidence | The listed HPA028943 antibody is Approved for IHC; HPA rates the overall tissue profile Approved with medium staining–RNA consistency (HPA: antibodies; HPA: tissue IHC). Neither rating establishes that every apparent positive cell is specific, so controls and morphology remain necessary. |
| Isoforms and target processing | UniProt lists two BUD31 isoforms, no signal peptide or propeptide, and one annotated protein chain spanning residues 1–144 (UniProt P41223). The supplied record gives no antibody epitope or isoform-specific staining evidence; do not assign a staining difference to either isoform. |
| Antigen retrieval | Retrieval is a routine variable in paraffin IHC, but no BUD31-specific retrieval condition or fixation-sensitivity result is supplied (general IHC practice). Follow the IHC-validated antibody's instructions and compare controls when optimizing; do not infer an epitope-masking mechanism from HPA staining levels. |
| Situation | Likely cause | Next action |
|---|---|---|
| The intended positive tissue has little or no chromogenic signal. | Detection failure or unsuitable assay conditions are possible; HPA reports medium staining in colon glandular cells (HPA: tissue IHC). | Check a detection control, tissue morphology and the antibody's IHC instructions; review retrieval and dilution as general IHC variables before interpreting the slide as BUD31-negative. |
| Signal appears only at cell borders or outside cells. | That distribution conflicts with UniProt's nuclear location and lacks support from HPA's cellular tissue pattern (UniProt P41223; HPA: tissue IHC). | Inspect a detection-only control and section morphology. Reassess staining conditions, and require convincing cellular signal in an HPA medium-staining tissue before assigning specificity. |
| A nominally negative comparison tissue stains strongly. | Cross-reactivity or endogenous detection activity is possible when staining conflicts with an HPA not-detected entry (HPA: tissue IHC). | Compare its cell type with the exact HPA entry, run a detection-only control, and inspect a positive comparison tissue; do not treat one HPA not-detected category as an absolute biological absence. |
| Brown haze makes nuclear scoring difficult. | Background from detection chemistry, insufficient washing or nonspecific binding may obscure the cell-resolved pattern (general IHC practice). | Check the detection-only control; optimize blocking, washes and detection conditions. Score only identifiable cells after background is reduced, using HPA's nuclear and cytoplasmic tissue pattern as context (HPA: tissue IHC). |
| The slide shows cytoplasmic staining but little nuclear detail. | HPA permits cytoplasmic tissue signal, while UniProt annotates the nucleus and HPA IF supports nucleoplasm as the main location (HPA: tissue IHC; UniProt P41223; HPA: subcellular ICC-IF). | Check nuclear counterstain and morphology, then compare an HPA medium-staining cell population. Record the compartment seen; do not call cytoplasmic staining alone conclusive evidence of either specificity or artifact. |
| IF/ICC shows puncta near the centrosome or signal along microtubules. | HPA lists centrosome and microtubule locations as uncertain; its supported main IF location is nucleoplasm (HPA: subcellular ICC-IF). | For the separate IF/ICC assessment, prioritize nucleoplasmic signal and treat additional patterns cautiously. HPA lists ICC support for HPA028943 (HPA: antibodies); that does not establish an IHC compartment pattern. |
Experiment evidence
IHC & IF images showing BUD31 staining
A11662 · IHCImmunohistochemistry (IHC) analyzes of BUD31 (E24) pAb in paraffin-embedded human tonsil carcinoma tissue at 1:50.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106245-BUD31/tissue/cerebral+cortex#imid_7593536
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106245-BUD31/tissue/colon#imid_7593462
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106245-BUD31/tissue/liver#imid_7593455
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106245-BUD31/tissue/kidney#imid_7593464
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028943; green = BUD31, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA028943; green = BUD31, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BUD31 IHC & IF
🧪Run appendix first: glandular cells should show BUD31 staining (HPA: Medium in appendix glandular cells). Use adrenal gland as the negative tissue, where glandular cells are listed as not detected (HPA: Not detected in adrenal gland glandular cells); on the appendix slide, cells without signal should show counterstain without specific nuclear chromogen, but their biological negative status is unverified.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BUD31 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality; use BUD31 knockout material or peptide blocking to assess target dependence (standard IHC practice). For chromogenic appendix IHC, quench endogenous peroxidase and check residual background in the no-primary control (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the selected A11662 paraffin-section caption does not state a fixative (caption: paraffin-embedded human tonsil carcinoma at 1:50; fixative unreported). Retrieval dependence is unreported, so compare retrieval conditions during IHC optimization (standard IHC practice). There is no supplied basis to say frozen sections or IF are easier; assess glandular background in appendix before scoring nuclear signal (HPA: Medium in appendix glandular cells; UniProt P41223: nucleus).
HPA tissue IHC evidence for BUD31
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in molecular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Soft tissue | Peripheral nerve | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BUD31 IHC Tips
Troubleshoot BUD31 staining in paraffin section IHC by checking retrieval, compartment, controls, and scoring before interpreting chromogenic signal.
How should I retrieve BUD31 in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page antigen retrieval setting). If nuclear staining is weak, compare a longer or shorter heating interval on adjacent sections while keeping the buffer constant, because retrieval can alter both epitope access and tissue morphology (standard IHC practice). Evaluate the change against nuclear signal in intact cells: BUD31 is annotated in the nucleus and at chromatin near androgen receptor target promoters (UniProt P41223). If optimization remains necessary, test another retrieval buffer only as a fallback, with matched controls and identical chromogenic development (standard IHC practice).
Could fixation explain weak or patchy BUD31 IHC staining?
BUD31 specific sensitivity to fixation is unknown from the supplied evidence; the selected 1:50 tissue image identifies paraffin sections but does not report a fixative (A11662 tissue IHC caption). Record the fixative, time before fixation, fixation duration, and section age for each specimen so staining differences can be assessed alongside specimen handling (standard IHC practice). Compare adjacent, similarly processed sections with the same retrieval and detection conditions before changing antibody concentration (standard IHC practice). Assess preserved nuclear morphology and staining in intact cells, since damaged or poorly processed areas can give misleading chromogenic results (standard IHC practice).
Should BUD31 staining be nuclear, cytoplasmic, or both?
Expect a substantial nuclear component because BUD31 is annotated in the nucleus, including chromatin at androgen receptor target promoters (UniProt P41223). Tissue IHC also reports cytoplasmic and nuclear expression across several tissues, with approved reliability and medium consistency with RNA data (HPA tissue IHC). Score nuclear and cytoplasmic signal separately in morphologically intact cells, using the same compartment definitions across specimens (standard IHC practice). If staining is exclusively cytoplasmic, inspect the matched negative control and detection background before interpreting it as BUD31; tissue IHC supports cytoplasmic signal, but the nuclear annotation warrants particular scrutiny (HPA tissue IHC; UniProt P41223; standard IHC practice).
Could isoforms or epitope masking explain discordant BUD31 staining?
BUD31 has 2 annotated isoforms, but the supplied tissue image does not identify the antibody epitope or establish isoform specific detection (UniProt P41223; A11662 tissue IHC caption). Its annotated protein chain spans residues 1–144, with no signal peptide, propeptide, transmembrane segment, or glycosylation sites (UniProt P41223). An annotated acetyllysine at residue 125 is a possible consideration only if the antibody epitope overlaps that site; overlap and any staining effect are unreported (UniProt P41223). Review the antibody epitope documentation, if available, and compare controlled retrieval conditions before attributing a staining difference to an isoform or modification (standard IHC practice).
How can companion IF help assess a BUD31 IHC pattern?
For a companion IF comparison, multiplex BUD31 with a marker for the cell population being scored, such as a glandular epithelial marker when examining glandular cells (HPA tissue IHC; standard IF practice). Include a nuclear counterstain to check whether signal overlaps nucleoplasm, the supported main IF location, while treating centrosome and microtubule assignments as uncertain (HPA subcellular). Choose a fluorophore channel with low specimen autofluorescence and inspect unstained tissue in that channel before interpreting weak signal (standard IF practice). Because BUD31 lacks a transmembrane segment and has nuclear localisation, permeabilise sufficiently to reach the nuclear side of cellular membranes, then check whether morphology remains intact (UniProt P41223; standard IF practice).
What should I check when BUD31 chromogenic staining looks diffuse?
Run a section without primary antibody and inspect endogenous pigment or enzyme signal before assigning diffuse chromogen to BUD31 (standard IHC practice). For peroxidase based detection, apply a peroxidase block and compare matched DAB development times; these are general workflow controls, not BUD31 specific validation (standard IHC practice). Check whether haze follows folds, section edges, drying marks, or necrotic areas, and exclude those regions from interpretation (standard IHC practice). The selected image documents staining at 1:50 in paraffin embedded human tonsil carcinoma tissue, but does not establish an optimal dilution for other specimens (A11662 tissue IHC caption).
How should I quantify BUD31 nuclear staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear scoring threshold before examining outcomes, then score only intact, evaluable cells in comparable regions (standard IHC practice). Report the percentage of positive nuclei and, when intensity varies, an H-score derived from the percentages at each intensity grade (standard IHC practice). Normalise positive nuclei to all evaluable nuclei in that same population; if reporting density per mm², use the measured evaluable tissue area as the denominator (standard IHC practice). Keep cytoplasmic staining separate because tissue IHC reports both compartments, and document threshold, counterstain, and batch conditions for comparisons (HPA tissue IHC; standard IHC practice).
When is apparent BUD31 positivity more likely an artefact?
Give greatest weight to reproducible staining in intact nuclei, consistent with BUD31 nuclear localisation and its reported role in pre-mRNA splicing (UniProt P41223). Cytoplasmic staining can occur in tissue IHC, so assess it separately; a signal confined to unexpected cells or damaged regions requires control review (HPA tissue IHC; standard IHC practice). Reject edge concentrated deposits, necrotic regions, and signal reproduced without primary antibody as evidence of specific expression (standard IHC practice). Check endogenous enzyme activity when using peroxidase and DAB, and interpret apparent negatives against tissue context because HPA reports medium staining in some cell populations and undetected staining in others (standard IHC practice; HPA tissue IHC).
Boster reagents
Best BUD31 / Protein BUD31 homolog IHC Antibodies
One anti-BUD31 antibody has real IHC data from paraffin-embedded human tonsil carcinoma tissue (A11662 image caption); its listed reactivity covers human, mouse, and rat (A11662 catalog).
Real IHC data
Anti-BUD31 (E24) Antibody
Cat # A11662
A11662 is the sole rendering SKU; it is listed for IHC and reacts with human, mouse, and rat samples (A11662 catalog). Its own IHC image shows paraffin-embedded human tonsil carcinoma tissue at 1:50 (A11662 image caption).
Which to pick: For tissue IHC, choose A11662, a rabbit polyclonal antibody listed at 1:50–1:200 (A11662 catalog); its image supports paraffin sections, while the fixative is unreported (A11662 image caption). No listed SKU has IF/ICC validation or an IF image (A11662 catalog). For cross-species work, A11662 lists human, mouse, and rat reactivity, although its IHC image shows human tissue only (A11662 catalog; A11662 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P41223 (BUD31_HUMAN, Protein BUD31 homolog).
- Human Protein Atlas. BUD31 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. BUD31 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the centrosome and microtubules..
- Human Protein Atlas. BUD31 antibody validation summary (1 antibodies).
- Downregulation of BUD31 Promotes Prostate Cancer Cell Proliferation and Migration via Activation of p-AKT and Vimentin In Vitro. International journal of molecular sciences 2023 — PMC10094631.
- The role of BUD31 in clear cell renal cell carcinoma: prognostic significance, alternative splicing, and tumor immune environment. Clinical and experimental medicine 2024 — PMC11322202.
- Splicing factor BUD31 promotes ovarian cancer progression through sustaining the expression of anti-apoptotic BCL2L12. Nature communications 2022 — PMC9587234.
- Single-cell proteo-genomic reveals a comprehensive map of centrosome-associated spliceosome components. iScience 2023 — PMC10214398.
- PubMed PMID:7841202 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12690205 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BUD31 (Protein BUD31 homolog). Expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A11662 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human tonsil carcinoma tissue; fixative not specified (datasheet A11662); 2. Fixation: Image fixative and duration unreported (datasheet A11662); 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-BUD31, 1:50-1:200 (datasheet A11662); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A11662); Caveat: Cytoplasmic staining may accompany the expected nuclear signal (HPA tissue IHC; UniProt); Isoform / epitope: 2 isoforms; antibody epitope coverage is unreported (UniProt; datasheet); Regulation: Staining intensity regulation is not established (UniProt). Evidence: UniProt P41223, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A11662, PMC10094631, PMC9587234. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BUD31/immunohistochemistry-ihc-guide.