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BCAS2 / Pre-mRNA-splicing factor SPF27 · IHC design guide
Design Immunohistochemistry for BCAS2
Plan chromogenic IHC-P for BCAS2 with the catalog antibody, starting at 5 μg/mL (datasheet: A07113). Assess nuclear and nucleolar staining, using high staining in testicular spermatogonia as a reference (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
BCAS2 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 nucleolar staining in tissue (HPA tissue IHC) | |
| Staining pattern | General nuclear and nucleolar staining (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Testis+4 more · see all | |
| Negative control | Adipose tissue |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin tissue fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07113) | |
| Caveat | Adipocytes may be unstained despite broad expression (HPA tissue IHC) | |
| Regulation | Ubiquitously expressed (UniProt) | |
| Isoform / epitope | No annotated isoforms; mature chain spans aa 2–225 (UniProt) |
Section 1
Recommended BCAS2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published chromogenic IHC protocols for pig testis, esophageal carcinoma, and human prostate tissue (PMC10607209; PMC5783606; PMC4453457).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A07113); 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-BCAS2, 5 μg/mL (datasheet A07113) |
| 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 | BCAS2-positive staining in spermatogonia cells of testis (HPA tissue IHC: High). HPA tissue profile: General nuclear and nucleolar expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 retrieval at 95–98 °C for 20 min for nuclear BCAS2; the published chromogenic protocols used citrate-based retrieval (page retrieval rule; PMC10607209; PMC5783606; PMC4453457).
Section 2
What Is the Expected BCAS2 Staining Pattern?
BCAS2 is expected chiefly in nuclei and nucleoli in paraffin-section IHC (HPA tissue IHC: general nuclear and nucleolar expression; UniProt O75934: nucleus and nucleolus). It is ubiquitously expressed, but staining varies by cell type (UniProt O75934: tissue specificity; HPA tissue IHC). HPA rates the tissue staining profile Supported, with medium consistency between antibody staining and RNA expression; use that qualification when interpreting weak or absent signal (HPA tissue IHC: reliability).
| Spermatogonia show strong nuclear or nucleolar staining; several other cell types show weaker staining. | This fits the reported range: High in testis spermatogonia and Medium in adrenal glandular cells, appendix lymphoid tissue, bronchial ciliated-cell bodies, and selected brain and cervical cells (HPA tissue IHC). Compare compartments as well as intensity; a positive cell should show the predominantly nuclear or nucleolar pattern reported for tissue IHC (HPA tissue IHC: profile). |
| The dominant stain outlines cell membranes or fills cytoplasm while nuclei remain clear. | Treat a predominantly membrane or cytoplasmic pattern as suspect for this IHC assay (UniProt O75934: nucleus and nucleolus; HPA tissue IHC: nuclear and nucleolar profile). BCAS2 has no annotated transmembrane segment or signal peptide (UniProt O75934: topology and processing). Check whether the same pattern persists in a negative reagent control and whether a known-positive section retains nuclear staining (standard IHC practice). |
| Adipocytes are strongly stained, especially when the expected nuclear pattern is absent. | Adipocytes were Not detected in the supplied HPA tissue IHC record, so strong staining there deserves scrutiny (HPA tissue IHC: adipocytes). Consider cross-reactivity or endogenous detection activity, especially if the colour does not track nuclei (standard IHC interpretation). This is a reference pattern, not proof that every adipocyte in every preparation must be blank (HPA tissue IHC: Supported, medium RNA-staining consistency). |
| Colour spreads across extracellular spaces or many cell compartments with little nuclear contrast. | Diffuse background does not reproduce the reported nuclear and nucleolar distribution (HPA tissue IHC: profile). Review the negative reagent control, blocking, washes, and detection development before assigning that colour to BCAS2 (standard IHC practice). Score specific nuclear signal separately from background; weak widespread colour alone does not establish protein-positive cells (standard IHC interpretation). |
| No nuclear staining appears in a known-positive testis section. | A blank spermatogonia population conflicts with the supplied High HPA observation (HPA tissue IHC: testis spermatogonia). First verify that the section contains identifiable spermatogonia, then inspect primary-antibody application and detection controls (standard IHC practice). The HPA result supports use of this tissue as a positive reference, but does not identify which preparation step failed (HPA tissue IHC; standard IHC practice). |
💡Expected BCAS2 appearanceCall the result positive when identifiable spermatogonia have High predominantly nuclear or nucleolar staining, while strong membrane, diffuse extracellular, or adipocyte staining prompts review as possible nonspecific signal (HPA tissue IHC: profile, spermatogonia and adipocytes; UniProt O75934: localisation; standard IHC interpretation).
| Tissue and cell selection | BCAS2 is ubiquitously expressed, yet HPA reports levels from High in spermatogonia to Not detected in adipocytes (UniProt O75934: tissue specificity; HPA tissue IHC). Identify the relevant cell population before comparing slides; a tissue label alone does not specify the expected intensity (HPA tissue IHC: cell-level observations). |
| Compartment and protein topology | Nuclear and nucleolar staining accords with the tissue profile and UniProt localisation (HPA tissue IHC; UniProt O75934). The record lists no transmembrane segment or signal peptide, offering no basis to expect a membrane or secreted-protein pattern (UniProt O75934: topology and processing). These annotations do not establish any BCAS2-specific fixation or antigen-retrieval effect. |
| Antibody evidence | The supplied HPA antibody HPA067881 has Supported IHC and Supported ICC status (HPA antibodies: HPA067881). HPA rates the tissue profile Supported with medium agreement between staining and RNA data (HPA tissue IHC: reliability). Treat this as support for the reported pattern while checking controls for the actual assay (standard IHC practice). |
| IF/ICC Q&A: where should BCAS2 appear? | In IF/ICC, look for nuclear speckles and a centrosome signal: HPA marks nuclear speckles Supported and centrosome Approved (HPA subcellular ICC-IF). This describes the separate IF/ICC application; the paraffin-section IHC reference remains general nuclear and nucleolar staining (HPA tissue IHC: profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive spermatogonia have no visible stain. | The result disagrees with HPA's High testis spermatogonia observation; the cause cannot be assigned from the staining record alone (HPA tissue IHC). | Confirm the expected cells are present, then check primary-antibody omission and detection-system controls; review the assay's validated retrieval and dilution settings (standard IHC practice). |
| Only cytoplasmic or membrane colour is seen. | The dominant compartment disagrees with BCAS2's nuclear and nucleolar annotation and the HPA tissue profile (UniProt O75934: localisation; HPA tissue IHC). | Check a known-positive section and a negative reagent control; interpret persistent nonnuclear colour cautiously (standard IHC practice). |
| Adipocytes stain strongly. | That finding conflicts with HPA's Not detected adipocyte entry and may reflect cross-reactivity or endogenous detection activity (HPA tissue IHC; standard IHC interpretation). | Compare staining with nuclear counterstain and negative reagent controls; investigate tissue-related detection background before scoring adipocytes positive (standard IHC practice). |
| The whole section has a faint, diffuse haze. | Poor contrast can obscure the expected nuclear pattern; the haze alone does not identify its source (HPA tissue IHC: profile; standard IHC interpretation). | Inspect negative controls, blocking, washes, and chromogen development, then judge whether discrete nuclear signal remains above background (standard IHC practice). |
| Adrenal gland or bronchus is weaker than testis. | Different intensities can match the reference: glandular cells of adrenal gland and ciliated-cell bodies of bronchus are Medium, versus High testis spermatogonia (HPA tissue IHC). | Compare the named cells and their nuclear pattern rather than requiring equal intensity across tissues (HPA tissue IHC; standard IHC interpretation). |
| A centrosome-like IF/ICC punctum seems inconsistent with IHC. | HPA's ICC-IF record includes centrosome as an Approved location alongside Supported nuclear speckles (HPA subcellular ICC-IF). | Interpret that observation using the separate IF/ICC localisation record; assess paraffin IHC against its nuclear and nucleolar tissue profile (HPA subcellular ICC-IF; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing BCAS2 staining
A07113 · IHCImmunohistochemistry of BCAS2 in human breast carcinoma with BCAS2 antibody at 5 μg/mL.
A07113 · IFImmunofluorescence of BCAS2 in human breast carcinoma tissue with BCAS2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000116752-BCAS2/tissue/cerebral+cortex#imid_18825435
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000116752-BCAS2/tissue/colon#imid_18825361
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000116752-BCAS2/tissue/liver#imid_18825346
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000116752-BCAS2/tissue/kidney#imid_18825355
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA067881; green = BCAS2, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA067881; green = BCAS2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for BCAS2 IHC & IF
🧪Run testis first and expect spermatogonia to stain (HPA: High in spermatogonia). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); no specific BCAS2-negative cell population is established within the testis slide, so assess unstained background against the no-primary control (UniProt: ubiquitously expressed; standard IHC practice).
Positive control tissue: Testis (Spermatogonia cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show BCAS2 in MCF-7, U-251MG, U2OS, with annotated localisation: Nuclear speckles (supported), Centrosome (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a concentration-matched isotype control matched to the primary antibody’s host species and clonality, and BCAS2-knockout material as a biological negative (standard IHC practice). Quench endogenous peroxidase for chromogenic detection, and block endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A BCAS2-specific fixation window and antigen-retrieval dependency are unreported in the supplied evidence; the A07113 breast-carcinoma IHC caption also leaves the fixative unreported (selected A07113 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier; ICC-IF images exist for MCF-7, U-251MG and U2OS, with nuclear speckles and centrosome reported as locations (HPA: subcellular). For testis chromogenic IHC, check for endogenous peroxidase background with the no-primary control (standard IHC practice).
HPA tissue IHC evidence for BCAS2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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 |
|---|---|---|---|---|
| Testis | Spermatogonia cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Bronchus | Ciliated cells (cell body) | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced BCAS2 IHC Tips
Troubleshoot BCAS2 staining in paraffin sections by checking retrieval, nuclear localisation, controls, and scoring before interpreting differences between specimens.
How should I retrieve BCAS2 in paraffin sections with weak nuclear staining?
Use Tris-EDTA at pH 9.0 for heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Keep section thickness, heating, cooling, and detection conditions consistent while comparing a weak section with a reference section (standard IHC practice). Score nuclear and nucleolar staining first, because those are the expected tissue compartments for BCAS2 (HPA tissue IHC: general nuclear and nucleolar expression). If staining remains weak, compare a second retrieval condition on adjacent sections as an optimisation experiment, recording tissue damage and background alongside signal (standard IHC practice).
Can I attribute weak BCAS2 staining to fixation?
BCAS2-specific fixation sensitivity is unknown from the supplied evidence; the selected antibody image identifies human breast carcinoma and 5 µg/mL antibody, but does not state its fixative (A07113 caption). Record the specimen’s fixative, fixation duration, and processing history before attributing a weak result to fixation (standard IHC practice). Compare adjacent sections with the same retrieval and detection conditions, using a reference specimen processed similarly whenever possible (standard IHC practice). Nuclear or nucleolar staining can guide assessment of a usable signal, but neither that pattern nor BCAS2 topology establishes a target-specific fixation effect (HPA tissue IHC: general nuclear and nucleolar expression; UniProt O75934 topology).
Which staining compartments support a BCAS2 result?
In tissue sections, assess BCAS2 primarily in nuclei and nucleoli (HPA tissue IHC: general nuclear and nucleolar expression; UniProt O75934: nucleus and nucleolus). Its spliceosome-associated function also makes a nuclear pattern biologically coherent (UniProt O75934: spliceosome component). The HPA subcellular record identifies nuclear speckles and centrosomes in cellular imaging, but a chromogenic section may not resolve those fine structures reliably (HPA subcellular: nuclear speckles and centrosome; standard microscopy practice). If staining is mainly diffuse cytoplasm or extracellular material, inspect counterstain alignment, section quality, and detection background before assigning it to BCAS2 (standard IHC practice).
How should epitope uncertainty affect BCAS2 IHC optimisation?
The supplied BCAS2 record lists 0 isoforms and a 225-aa protein without a transmembrane segment or annotated glycosylation sites (UniProt O75934 record). It also lists N-terminal acetylation at residue 2 and phosphorylation at residue 94, while the antibody’s binding epitope is unspecified (UniProt O75934 record; A07113 caption). These annotations do not establish whether either modification changes staining with this antibody (UniProt O75934 record; A07113 caption). When signal changes after retrieval, compare matched sections and controls before attributing the change to an isoform or modification (standard IHC practice).
How can I check BCAS2 by IF alongside chromogenic IHC?
For a separate IF experiment, multiplex BCAS2 with a marker identifying the expected cell population, such as spermatogonia in testis, and verify that the BCAS2 signal occupies the appropriate nuclei (HPA tissue IHC: high in spermatogonia; HPA tissue IHC: general nuclear expression). Select fluorophores after checking specimen autofluorescence and include single-channel controls to detect bleed-through (standard IF practice). If the antibody recognises an intranuclear epitope, optimise permeabilisation to admit antibody across the nuclear envelope while preserving nuclear detail; its epitope is unspecified here (A07113 caption; standard IF practice). Compare localisation across methods, while assessing IF fixation and antibody performance independently of the tissue-IHC image (A07113 caption; standard IF practice).
What should I adjust when BCAS2 staining has diffuse background?
Compare no-primary and reagent-only controls with the stained section to locate background from the detection system or tissue (standard chromogenic IHC practice). For peroxidase-based detection, check the peroxidase block and development time, then titrate the primary antibody and wash stringency on matched sections (standard chromogenic IHC practice). The selected BCAS2 tissue image reports 5 µg/mL antibody in human breast carcinoma, but provides no fixation or detection details (A07113 caption). Preserve interpretable nuclear and nucleolar signal during optimisation, since diffuse colour outside those compartments is a poor basis for BCAS2 scoring (HPA tissue IHC: general nuclear and nucleolar expression; standard IHC practice).
How should I quantify BCAS2 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and nuclear scoring rule before examining outcomes, then report the percentage of positive nuclei and an H-score if intensity is reproducible (standard IHC scoring practice). Normalise positive counts to the number of evaluable cells in that population, or report positive-cell density per mm² of evaluable tissue when area is the denominator (standard IHC scoring practice). Exclude necrotic, folded, and poorly counterstained regions using the same criteria in every section (standard IHC scoring practice). Record nuclear and nucleolar signal separately if the question requires it, because both compartments are reported for BCAS2 (HPA tissue IHC: general nuclear and nucleolar expression).
How can I distinguish genuine BCAS2 staining from artefact?
A credible result shows staining in the expected nuclei or nucleoli, with cell-by-cell distribution that remains interpretable against the counterstain (HPA tissue IHC: general nuclear and nucleolar expression; standard IHC practice). Compare the observed cell population with the specimen: HPA reports high staining in testis spermatogonia, whereas adipocytes are listed as not detected (HPA tissue IHC: spermatogonia high; adipocytes not detected). Treat staining confined to section edges, necrosis, or predominantly extracellular space as suspect until controls and tissue morphology support it (standard IHC practice). For peroxidase detection, check a no-primary control and the peroxidase block before interpreting residual brown colour as BCAS2 (standard chromogenic IHC practice).
Boster reagents
Best BCAS2 / Pre-mRNA-splicing factor SPF27 IHC Antibodies
Anti-BCAS2 A07113 has IHC and IF images from human breast carcinoma (catalog image captions); its listed reactivity is human, mouse and rat (catalog: reactivity).
Real IHC data
Anti-Pre-mRNA-splicing factor SPF27 BCAS2 Antibody
Cat # A07113
A07113 will render with its own IHC image of human breast carcinoma at 5 μg/mL (catalog: A07113 IHC image caption). Its IF image shows human breast carcinoma tissue at 20 μg/mL (catalog: A07113 IF image caption).
Which to pick: Choose A07113 for tissue IHC: IHC-P is listed, and its own IHC image shows human breast carcinoma (catalog: A07113 applications and IHC image caption); the fixative is unreported (catalog: A07113 IHC image caption). For IF, A07113 has a tissue image; for ICC, A07113-1 lists ICC but has no supplied ICC image (catalog: A07113 IF image caption; A07113-1 applications and image alts). Both list human, mouse and rat reactivity, but the supplied IHC and IF images show human samples only; clone type is unreported for both (catalog: reactivity, image captions and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75934 (SPF27_HUMAN, Pre-mRNA-splicing factor SPF27).
- Human Protein Atlas. BCAS2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. BCAS2 subcellular location (ICC-IF): Localized to the nuclear speckles and centrosome..
- Human Protein Atlas. BCAS2 antibody validation summary (1 antibodies).
- Molecular characterization, expression patterns and cellular localization of BCAS2 gene in male Hezuo pig. PeerJ 2023 — PMC10607209.
- miR-486 functions as a tumor suppressor in esophageal cancer by targeting CDK4/BCAS2. Oncology reports 2018 — PMC5783606.
- BCAS2 promotes prostate cancer cells proliferation by enhancing AR mRNA transcription and protein stability. British journal of cancer 2015 — PMC4453457.
- BCAS2 is involved in alternative mRNA splicing in spermatogonia and the transition to meiosis. Nature communications 2017 — PMC5290162.
- PubMed PMID:9731529 — UniProt-cited evidence.
- PubMed PMID:10403562 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for BCAS2 (Pre-mRNA-splicing factor SPF27). Expected localisation Nuclear and nucleolar staining in tissue (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A07113 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Spermatogonia 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 A07113); 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-BCAS2, 5 μg/mL (datasheet A07113); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin tissue fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07113); Caveat: Adipocytes may be unstained despite broad expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; mature chain spans aa 2–225 (UniProt); Regulation: Ubiquitously expressed (UniProt). Evidence: UniProt O75934, HPA tissue IHC · Supported, HPA ICC-IF · nuclear speckles and centrosome., Datasheet A07113, PMC10607209, PMC5783606. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/BCAS2/immunohistochemistry-ihc-guide.