This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
BCAS2 / Pre-mRNA-splicing factor SPF27 · IHC design guide
Design Immunohistochemistry for BCAS2
Plan chromogenic BCAS2 IHC-P around nuclear and nucleolar tissue staining (HPA tissue IHC). Testis spermatogonia provide a high-staining reference (HPA tissue IHC); start the IHC-validated antibody A07113 at 5 μg/mL (datasheet: IHC-P).
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 (HPA tissue IHC) | |
| Staining pattern | Nuclear and nucleolar staining across tissue cell types (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 formalin fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes may be unstained despite the general tissue pattern (HPA tissue IHC) | |
| Regulation | Ubiquitously expressed (UniProt) | |
| Isoform / epitope | No isoforms; the annotated chain starts at residue 2 (UniProt) |
Section 1
Recommended BCAS2 IHC & IF Protocols
The catalog antibody protocol is followed by three published BCAS2 IHC protocols for pig testis, esophageal carcinoma tissue, and human prostate specimens (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 the page’s Tris-EDTA pH 9.0 retrieval (page retrieval rule); citrate retrieval is a published alternative (PMC10607209; PMC4453457).
Section 2
What Is the Expected BCAS2 Staining Pattern?
BCAS2 is expected in nuclei and nucleoli (UniProt O75934; HPA: general nuclear and nucleolar expression). Testis spermatogonia show high staining, while several other cell populations show medium staining (HPA: tissue IHC). Tissue IHC reliability is Supported, with medium consistency between staining and RNA data (HPA: reliability). BCAS2 has no transmembrane segment (UniProt O75934 topology).
| Distinct nuclear staining, sometimes accenting nucleoli, in spermatogonia or other HPA-positive cells. | This matches the reported compartment and cell distribution (UniProt O75934: nucleus and nucleolus; HPA: general nuclear and nucleolar expression). Spermatogonia are a high-staining reference; adrenal glandular cells and cerebral cortical neurons are medium-staining examples (HPA: tissue IHC). Judge cells within their tissue context rather than expecting every nucleus to have equal intensity. |
| Predominantly cytoplasmic or membranous chromogen with little nuclear signal. | The dominant compartment conflicts with BCAS2’s reported nuclear and nucleolar location (UniProt O75934; HPA: tissue IHC). A membrane pattern also lacks support from its topology (UniProt O75934: no transmembrane segment). Treat it as suspect staining and check localisation in a known-positive tissue before interpreting it as BCAS2. |
| Strong staining in adipocytes, especially if nuclei in a known-positive area remain weak. | HPA reports BCAS2 as not detected in adipocytes (HPA: adipose tissue IHC). This discrepancy can reflect nonspecific antibody binding or endogenous detection activity; it does not by itself identify which occurred. Compare primary-antibody and no-primary controls, and assess whether any chromogen is actually nuclear (General IHC practice). |
| A diffuse haze covers cells, stroma, and empty areas without clear nuclear boundaries. | That appearance cannot support a BCAS2 localisation call because the expected tissue pattern is nuclear and nucleolar (HPA: tissue IHC; UniProt O75934). Excess background can arise from nonspecific binding or detection chemistry (General IHC practice). Score only distinguishable cellular signal after the background has been resolved. |
| No nuclear signal in a section containing identifiable spermatogonia. | This conflicts with the high staining reported for spermatogonia (HPA: testis IHC), but a failed assay is only one possibility. First confirm that the relevant cells are present and interpretable, then review the antibody and detection controls (General IHC practice). HPA’s Supported reliability does not guarantee positivity in every specimen (HPA: reliability). |
💡Expected BCAS2 appearanceCall a positive result when spermatogonia show clear, relatively strong nuclear or nucleolar chromogen (HPA: high in spermatogonia; HPA: general nuclear and nucleolar expression); predominantly cytoplasmic, membranous, or diffuse acellular colour is suspect against that reference (UniProt O75934: nucleus, nucleolus, and no transmembrane segment).
| Tissue and cell selection | BCAS2 is described as ubiquitously expressed (UniProt O75934), yet HPA staining varies by cell type: high in spermatogonia, medium in selected glandular, lymphoid, ciliated, glial, neuronal, and squamous cells, and not detected in adipocytes (HPA: tissue IHC). Use the named cell population when comparing signal; tissue-wide averages can obscure the pattern. |
| Evidence strength and antibody choice | The tissue IHC profile is Supported, with medium consistency against RNA data; the listed HPA067881 antibody has Supported IHC validation (HPA: reliability; HPA: antibody validation). These labels support a reference pattern but do not establish identical staining with every IHC-validated antibody or every specimen. |
| Compartment and molecular context | UniProt places BCAS2 in the nucleus and nucleolus and describes a spliceosome role (UniProt O75934: subcellular location and function). It records no transmembrane segment, signal peptide, or propeptide (UniProt O75934: topology and processing). These facts support a nuclear readout; they do not predict an antibody epitope or antigen-retrieval requirement. |
| IF/ICC Q&A: Should punctate nuclear signal be expected? | Yes, nuclear speckles are the supported main IF/ICC location, and centrosome is an approved location (HPA: subcellular ICC-IF). Those cell-image findings provide context for IF/ICC interpretation; they do not require a punctate chromogenic pattern in paraffin tissue, whose reported profile is general nuclear and nucleolar expression (HPA: tissue IHC). |
| Target-specific fixation sensitivity | No target-specific fixation effect is established by the supplied UniProt or HPA records. Select and assess retrieval and detection conditions as general IHC workflow variables (General IHC practice); do not infer a BCAS2 fixation response from its topology, modifications, or tissue staining levels. |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or absent signal in identifiable spermatogonia. | The result differs from the reported high level in those cells (HPA: testis IHC); the source does not identify a BCAS2-specific cause. | Check cell preservation, primary-antibody application, detection reagents, and a known-positive control; review retrieval as a general IHC variable (General IHC practice). |
| Chromogen is mainly cytoplasmic or membranous. | That distribution conflicts with the nuclear and nucleolar reference and lacks topology support for a membrane pattern (UniProt O75934; HPA: tissue IHC). | Compare with a known-positive section and a no-primary control; only score a cellular nuclear pattern as supportive (General IHC practice; HPA: tissue IHC). |
| Adipocytes appear strongly positive. | They are reported as not detected (HPA: adipose tissue IHC); nonspecific binding or endogenous detection activity is possible (General IHC practice). | Inspect subcellular placement and no-primary controls, then reassess blocking and detection background (General IHC practice). |
| Colour appears in stroma or across the whole section. | Diffuse signal obscures the expected nuclear and nucleolar profile (HPA: tissue IHC); background may arise during antibody or chromogen detection (General IHC practice). | Compare a no-primary control, check washing and blocking, and adjust detection conditions if background persists (General IHC practice). |
| Only some cell types stain within an otherwise interpretable section. | Cell-type variation is compatible with the HPA range from high spermatogonia to medium, low, or undetected populations (HPA: tissue IHC). | Identify the cells before scoring and compare each with its HPA cell-specific reference; do not require equal intensity across a tissue (HPA: tissue IHC). |
| IF/ICC speckles are seen, but the paraffin section lacks discrete puncta. | HPA describes nuclear speckles in ICC-IF and general nuclear and nucleolar expression in tissue IHC (HPA: subcellular ICC-IF; HPA: tissue IHC). | Interpret the paraffin section against its tissue IHC reference: assess nuclear localisation and the named positive cells without requiring resolvable speckles (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 score spermatogonia for staining (HPA: High in spermatogonia). Use adipose tissue as the negative comparator; adipocytes are listed as not detected (HPA: adipocytes Not detected). Within the testis slide, unstained cells should show only counterstain or background, but no cell type is established here as an internal biological negative (HPA: testis spermatogonia High; UniProt O75934: ubiquitously expressed).
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 and an isotype control matched to the primary antibody’s host species, clonality, and concentration (standard IHC practice). If available, use BCAS2 knockout tissue or immunizing-peptide competition as a biological specificity control, and block endogenous peroxidase before chromogenic detection in testis (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval dependence are unreported in the supplied evidence; optimize antigen retrieval empirically for paraffin sections (supplied evidence; standard IHC practice). The selected A07113 breast-carcinoma IHC caption reports 5 μg/mL but leaves the fixative unreported (A07113 tissue-IHC caption). No supplied comparison establishes whether frozen sections or IF are easier, and no testis-specific artefact is reported (supplied evidence).
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
Troubleshooting BCAS2 staining in paraffin sections centers on retrieval, nuclear localisation and controls that distinguish signal from tissue artefact.
What retrieval should I try when BCAS2 staining is weak in paraffin sections?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval specification). Allow sections to cool in buffer before washing, and run the same retrieval on control and experimental sections so staining intensity remains comparable (standard IHC practice). BCAS2 is a nuclear and nucleolar protein (UniProt O75934); assess recovery in nuclei rather than treating diffuse cytoplasmic colour as improved retrieval. If nuclear staining remains weak, compare a milder or alternative retrieval condition on serial sections while keeping antibody concentration and detection constant (standard IHC practice).
Could fixation explain weak or uneven BCAS2 staining?
Target-specific fixation sensitivity is unknown from the supplied evidence; record the fixative, fixation duration and processing history before changing the staining protocol. The breast carcinoma image for A07113 reports BCAS2 staining at 5 µg/mL, but its caption does not state the fixative (A07113 tissue-IHC caption). For paraffin IHC, compare sections with documented processing under the same pH 9.0, 20 min retrieval condition and detection settings (page retrieval specification; standard IHC practice). If only tissue edges stain strongly, inspect processing and section quality before attributing the pattern to BCAS2 biology (standard IHC practice).
How should I judge nuclear, nucleolar or cytoplasmic BCAS2 staining?
Score convincing signal in nuclei, including nucleoli, because those compartments are reported for BCAS2 (UniProt O75934; HPA tissue IHC). Examine the nuclear pattern at high magnification against the counterstain, then record any nucleolar enrichment separately from overall nuclear positivity (standard IHC practice). BCAS2 has no annotated transmembrane segment (UniProt O75934 topology), so a dominant membrane rim is a reason to investigate staining specificity. HPA also reports nuclear speckles and a centrosome location in cell imaging (HPA subcellular); do not assign punctate chromogenic deposits to either structure without adequate resolution and independent validation (standard IHC practice).
Can BCAS2 isoforms or modifications explain discordant staining?
The supplied record lists 0 isoforms and no annotated domains, so isoform-specific staining cannot be inferred from it (UniProt O75934). It does list N-terminal acetylation at residue 2 and phosphorylation at serine 94 (UniProt O75934 modified residues). Whether either modification affects this antibody is unknown because its epitope is not supplied; avoid assigning staining differences to a modification without epitope or competition evidence. Compare sections with matched retrieval and detection, and use an independently validated BCAS2 reagent if epitope masking or specificity remains in question (standard IHC practice).
How can I check BCAS2 localisation by multiplex IF?
For a separate IF experiment, pair BCAS2 with a validated marker for the expected cell type; spermatogonia show high BCAS2 staining in testis IHC (HPA tissue IHC). Choose a spectrally separated, preferably far-red BCAS2 channel when tissue autofluorescence overlaps shorter wavelengths, and include single-colour controls (standard IF practice). BCAS2 is nuclear and has no transmembrane segment (UniProt O75934), so permeabilise sufficiently to admit antibody across cellular and nuclear barriers while preserving nuclear structure (standard IF practice). Compare nuclear signal with a DNA counterstain and use no-primary controls to identify background before interpreting puncta (standard IF practice).
What should I check when BCAS2 IHC shows widespread brown background?
First inspect a no-primary control and the section’s unstained pigment; either can reveal colour unrelated to antibody binding (standard IHC practice). For peroxidase and DAB detection, apply an appropriate peroxidase block and assess endogenous enzyme activity in the control section (standard chromogenic IHC practice). Reduce nonspecific signal by checking blocking, washes, primary concentration and detector exposure one variable at a time; the A07113 image caption reports 5 µg/mL only for that image (A07113 tissue-IHC caption; standard IHC practice). Interpret diffuse cytoplasmic colour cautiously because the reported tissue pattern is chiefly nuclear and nucleolar (HPA tissue IHC).
What is a defensible way to quantify BCAS2 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define cell classes and tissue regions before scoring, then report nuclear % positive cells or a nuclear H-score using fixed intensity categories (standard IHC practice). Normalise positive counts to the number of evaluable nuclei in each annotated region, and exclude folds, necrosis and tissue edges using the same rules across sections (standard IHC practice). If counting discrete positive cells per mm², report the analysed tissue area and cell class so density is interpretable (standard IHC practice). Record nucleolar enrichment separately where visible because BCAS2 has a reported nuclear and nucleolar tissue pattern (HPA tissue IHC).
How do I distinguish true BCAS2 positivity from staining artefact?
A credible BCAS2 pattern is predominantly nuclear or nucleolar, consistent with the supplied localisation and tissue IHC profile (UniProt O75934; HPA tissue IHC). Check whether the expected cell population stains: HPA reports high signal in testis spermatogonia and no detected signal in adipocytes (HPA tissue IHC). Reject a biological interpretation of staining confined to cut edges, necrotic areas or obvious pigment until those features are excluded in control sections (standard IHC practice). Diffuse membrane staining is discordant with the reported nuclear pattern, while a positive no-primary control suggests endogenous enzyme activity or detection background (HPA tissue IHC; standard chromogenic IHC practice).
Boster reagents
Best BCAS2 / Pre-mRNA-splicing factor SPF27 IHC Antibodies
A07113 has human breast carcinoma IHC and IF images (catalog image captions); its listed reactivity includes human, mouse and rat (catalog applications and reactivity).
Real IHC data
Anti-Pre-mRNA-splicing factor SPF27 BCAS2 Antibody
Cat # A07113
A07113 is the only SKU rendered, with IHC data from human breast carcinoma at 5 μg/mL (A07113 IHC image caption). Its IF image also shows human breast carcinoma tissue at 20 μg/mL (A07113 IF image caption).
Which to pick: Choose A07113 for paraffin-section IHC because it lists IHC-P and has its own human breast carcinoma IHC image (A07113 applications; A07113 IHC image caption); the fixative is unreported (A07113 IHC image caption). Choose A07113 for tissue IF with an image, or A07113-1 when an ICC-listed antibody is needed; A07113-1 has no IHC image or IHC-P listing (A07113 IF image caption; A07113-1 applications and image captions). Both list human, mouse and rat reactivity, but the supplied IHC and IF images show human tissue only (catalog reactivity; A07113 image captions).
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 (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 formalin fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Adipocytes may be unstained despite the general tissue pattern (HPA tissue IHC); Isoform / epitope: No isoforms; the annotated chain starts at residue 2 (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.