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CBFA2T3 / Transcriptional corepressor CBFA2T3 · IHC design guide
Design Immunohistochemistry for CBFA2T3
Plan CBFA2T3 paraffin IHC around the general nuclear tissue staining reported by HPA (HPA tissue IHC). This guide covers fixation consistency, the catalog antibody’s 1:100–1:300 IHC dilution (datasheet), chromogenic detection and interpretation of staining that may diverge from RNA levels (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CBFA2T3 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 | General nuclear staining in tissue (HPA tissue IHC) | |
| Staining pattern | Nuclear staining in adipocytes and glandular cells (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 | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
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 | Staining may diverge from tissue RNA levels (HPA tissue IHC) | |
| Regulation | Brain, lymphoid tissue and pancreas have enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | Four isoforms; verify epitope coverage across variants (UniProt) |
Section 1
Recommended CBFA2T3 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet) is accompanied by published CBFA2T3 IHC conditions for osteosarcoma (PMC4467124) and T-cell lymphoma (PMC7002219).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A30478) |
| Fixation | Image fixative and duration unreported (datasheet A30478); 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-CBFA2T3, 1:100 - 1:300 (datasheet A30478) |
| 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 | CBFA2T3-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: General nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval); compare citrate pH 6.0 if needed (PMC4467124).
Section 2
What Is the Expected CBFA2T3 Staining Pattern?
CBFA2T3 should be assessed primarily as nuclear staining in paraffin sections: HPA reports general nuclear expression and high staining in several cell types, including adipocytes, glandular cells, respiratory epithelial cells, glial cells and Purkinje cells (HPA tissue IHC). UniProt also lists nucleolar, nucleoplasmic and Golgi membrane locations, with no transmembrane segment (UniProt O75081 topology). HPA rates tissue IHC staining Approved but reports low consistency with RNA expression (HPA tissue IHC).
| Distinct nuclear staining in adipocytes, glandular cells or respiratory epithelial cells. | This fits the reported general nuclear profile and High staining in those cell types (HPA tissue IHC). Score the relevant cells and nuclei on the section; HPA's High category does not prescribe a numerical intensity cutoff (HPA tissue IHC). |
| Staining appears chiefly cytoplasmic or outlines cell membranes, with little nuclear signal. | That does not match the general nuclear IHC profile (HPA tissue IHC). UniProt lists a Golgi membrane location, so a minor extranuclear component alone is inconclusive; predominant membrane outlining warrants a specificity and detection review (UniProt O75081 subcellular location; general IHC practice). |
| Strong staining appears in a cell population outside the listed High examples. | Do not classify that population as negative solely from this excerpt: the HPA payload lists selected High findings and provides no negative-cell list (HPA tissue IHC). Check cell identity, nuclear localisation and controls before considering cross-reactivity or endogenous detection activity (general IHC practice). |
| Color spreads across nuclei and surrounding tissue, obscuring cell boundaries. | The distribution cannot be scored reliably as CBFA2T3 when nuclear enrichment is unclear (HPA tissue IHC; general IHC practice). Background from blocking, washing or chromogen detection should be assessed with appropriate controls (general IHC practice). |
| No nuclear signal appears in a section expected to contain a listed High cell type. | This conflicts with the reported High observation for that cell type, but a single absent stain does not establish absent protein (HPA tissue IHC). Review cell identity, section quality and assay performance; HPA reports low staining–RNA consistency (HPA tissue IHC; general IHC practice). |
💡Expected CBFA2T3 appearanceA convincing positive result is nuclear staining in an HPA-listed High cell type, such as an adipocyte or glandular cell; widespread non-nuclear color alone is suspect (HPA tissue IHC; general IHC practice).
| Which compartment carries the main interpretive weight? | General nuclear expression is the tissue IHC profile (HPA tissue IHC). UniProt includes nucleolus, nucleoplasm and Golgi membrane, but those annotations do not make predominantly membranous chromogen staining a validated tissue pattern (UniProt O75081 subcellular location; HPA tissue IHC). |
| How should tissue and cell selection be interpreted? | HPA reports High staining in adipocytes, adrenal and appendix glandular cells, bronchial respiratory epithelium, caudate glia, cerebellar Purkinje cells and cerebral cortical endothelium (HPA tissue IHC). These are observed examples, not a complete positive–negative panel (HPA tissue IHC). |
| How strong is the tissue IHC evidence? | The tissue profile is Approved, while its antibody staining has low consistency with RNA expression data (HPA tissue IHC). HPA065890 is Approved for IHC in the supplied antibody list (HPA antibodies); interpret unexpected tissue intensity with morphology and controls rather than RNA ranking alone (general IHC practice). |
| Do isoforms or processing specify different stain patterns? | UniProt lists four isoforms, one annotated chain spanning residues 1–653, and no signal peptide or propeptide (UniProt O75081). The supplied record does not map antibody epitopes or establish isoform-specific IHC staining, so those features cannot explain a particular positive or negative section (UniProt O75081). |
| What should IF/ICC show? | HPA reports enhanced nucleoplasmic localisation in ICC-IF images from MCF-7, SiHa and U2OS (HPA subcellular). HPA059931 and HPA062423 have Enhanced ICC status, while the listed IHC antibody has Approved IHC status (HPA antibodies). Use the separate IF/ICC guide for its assay design. |
| Situation | Likely cause | Next action |
|---|---|---|
| Listed High cells show no nuclear chromogen signal. | The expected HPA observation is absent, or the assay failed; this result alone cannot distinguish those possibilities (HPA tissue IHC; general IHC practice). | Confirm the listed cell type is present, then check antibody use, retrieval, detection and a positive control under the assay's instructions (general IHC practice). |
| Signal is predominantly cytoplasmic or membrane-like. | This diverges from general nuclear tissue IHC staining, although UniProt lists a Golgi membrane location (HPA tissue IHC; UniProt O75081). | Recheck morphology and nuclear counterstain; compare a positive control and a primary-omission control before assigning specificity (general IHC practice). |
| Diffuse color obscures nuclear detail. | Background or excessive detection can prevent assessment of the HPA nuclear pattern (HPA tissue IHC; general IHC practice). | Inspect blocking, wash and detection controls, then adjust the general IHC workflow according to the assay instructions (general IHC practice). |
| A surprising cell population stains strongly. | The supplied HPA extract has no negative-cell list, so an unlisted population is not automatically erroneous (HPA tissue IHC). | Confirm the cell type and nuclear pattern; use appropriate negative controls to investigate cross-reactivity or endogenous detection activity (general IHC practice). |
| Staining intensity disagrees with tissue RNA expectations. | HPA explicitly reports low consistency between antibody staining and RNA expression, despite Approved tissue IHC reliability (HPA tissue IHC). | Record the observed cell-level stain and controls; avoid treating RNA abundance as an IHC intensity standard (HPA tissue IHC; general IHC practice). |
| IF/ICC appears nucleoplasmic but the paraffin IHC result is unclear. | The enhanced IF/ICC localisation and Approved tissue IHC status refer to different antibody validation entries and assay contexts (HPA antibodies; HPA subcellular). | Evaluate the IHC section with its own morphology and controls; consult the separate IF/ICC guide for fluorescence assay decisions (general IHC practice). |
Experiment evidence
IHC & IF images showing CBFA2T3 staining
A30478 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using MTG16 Antibody. The picture on the right is blocked with the sy
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000129993-CBFA2T3/tissue/cerebral+cortex#imid_19949525
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000129993-CBFA2T3/tissue/colon#imid_19949451
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000129993-CBFA2T3/tissue/liver#imid_19949460
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000129993-CBFA2T3/tissue/kidney#imid_19949453
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA059931; green = CBFA2T3, blue = nucleus, red = microtubules.
HPA · ICC-IF MCF-7Human Protein Atlas immunofluorescence, MCF-7, antibody HPA059931; green = CBFA2T3, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CBFA2T3 IHC & IF
🧪Run adipose tissue first and assess staining in adipocytes (HPA: High in adipocytes). HPA detects CBFA2T3 in all 45 scored tissues, so use no-primary and isotype controls in place of a negative tissue; cells without specific staining on the positive slide can indicate background, but should not be designated biological negatives without validation (HPA: no negative tissue rows; standard IHC practice).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: CBFA2T3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CBFA2T3 in MCF-7, SiHa, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and an isotype control matched to the primary antibody’s host species and clonality; use knockout tissue or a validated peptide-block condition to test specificity (standard IHC practice; A30478 tissue-IHC caption: peptide-block image). Quench endogenous peroxidase before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of antigen retrieval are unreported in the supplied evidence; optimize retrieval for the IHC-validated antibody (standard IHC practice). The selected A30478 paraffin-section caption does not state a fixative (A30478 tissue-IHC caption: fixative unreported). The evidence does not establish whether frozen sections or IF are easier; in adipose sections, lipid extraction can leave empty vacuoles that complicate cell identification (standard histology practice).
HPA tissue IHC evidence for CBFA2T3
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Breast | Adipocytes | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: CBFA2T3 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced CBFA2T3 IHC Tips
Troubleshoot CBFA2T3 staining in paraffin sections by checking retrieval, nuclear localisation, cell type and matched controls before scoring chromogenic signal.
How should I adjust retrieval when CBFA2T3 nuclear staining is weak?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval rule: nuclear antigen). If nuclear staining remains weak, compare a shorter heating time or a citrate buffer at pH 6.0 on adjacent sections as a fallback experiment (standard IHC practice). Keep section thickness, the catalog antibody concentration, detection chemistry and development time constant so the comparison tests retrieval rather than several changes at once (standard IHC practice). Score nucleoplasmic signal in the same cell type across conditions (HPA: enhanced nucleoplasmic localisation), while checking whether morphology or diffuse staining deteriorates (standard IHC practice).
Could fixation explain variable CBFA2T3 staining between paraffin blocks?
Target-specific fixation sensitivity is unknown; the selected catalog antibody image shows paraffin-embedded human brain, but its caption does not report a fixative (catalog antibody A30478: tissue-IHC caption). For a prospective comparison, process matched specimens with documented fixation conditions, such as 10% neutral buffered formalin for 6 versus 24 hours, followed by the same embedding and staining workflow (standard IHC practice). Compare nuclear signal, tissue morphology and background within the same cell type, recording block age and processing history (standard IHC practice; HPA: general nuclear expression). Treat any difference as an observation from that comparison rather than a known CBFA2T3 fixation effect (catalog antibody A30478: fixative unreported).
How should I assess cytoplasmic or punctate CBFA2T3 staining?
Prioritise nucleoplasmic staining when assessing the result, because the subcellular profile identifies enhanced nucleoplasmic localisation (HPA: subcellular profile). The protein record also lists nucleus, nucleolus and Golgi apparatus membrane, with no transmembrane segment, so a punctate pattern merits careful compartment assessment rather than automatic rejection (UniProt O75081: localisation and topology). Compare chromogenic signal with the nuclear counterstain in intact cells and inspect adjacent sections stained without primary antibody (standard IHC practice). Document nuclear and extranuclear patterns separately, and require reproducibility plus an appropriate antibody control before assigning biological meaning to an unexpected compartment (standard IHC practice; catalog antibody A30478: peptide-blocked image).
Can epitope choice change which CBFA2T3-positive cells I detect?
CBFA2T3 has 4 recorded isoforms, but the supplied catalog image does not identify the antibody epitope or establish which isoforms it detects (UniProt O75081: isoforms; catalog antibody A30478: tissue-IHC caption). Obtain the immunogen or epitope coordinates before interpreting staining as total CBFA2T3, then compare those coordinates with the isoform sequences (standard IHC practice; UniProt O75081: isoforms). The TAFH domain spans residues 171–266, and recorded phosphorylation sites include 457, 459 and 479 (UniProt O75081: domains and modified residues). If the epitope overlaps a variable or modified region, test an independent epitope and report antibody identity with the scoring results (standard IHC practice).
How can IF help check a questionable chromogenic CBFA2T3 pattern?
Use IF as a separate validation experiment: the subcellular profile reports enhanced nucleoplasmic localisation and lists ICC/IF images in MCF-7, SiHa and U2OS cells (HPA: subcellular profile). In tissue, multiplex CBFA2T3 with a marker for the expected cell type, such as a glial-cell marker when examining caudate, and include a nuclear counterstain (HPA: High in caudate glial cells; standard IF practice). Choose a far-red channel when tissue autofluorescence obscures shorter wavelengths, and include single-channel and secondary-only controls (standard IF practice). For a nucleoplasmic epitope, trial mild permeabilisation, such as 0.1% Triton X-100 for 5 min; the membrane-facing orientation of the reported Golgi-associated pool is unspecified (standard IF practice; UniProt O75081: localisation and topology).
What controls distinguish CBFA2T3 signal from chromogenic background?
Run a no-primary section and inspect pigment, folds, edges and necrotic areas before judging weak nuclear colour (standard IHC practice). For peroxidase detection, a 3% hydrogen peroxide block for approximately 10 min and controlled DAB development are general workflow steps, not evidence of CBFA2T3 specificity (standard IHC practice). The selected paraffin-brain image includes a synthesized-peptide-blocked comparison, which can inform specificity for that catalog antibody under the pictured conditions (catalog antibody A30478: tissue-IHC caption). If background persists, adjust blocking and antibody concentration one variable at a time, then compare intact nuclei in matched tissue regions (standard IHC practice; HPA: general nuclear expression).
How should I quantify CBFA2T3 across mixed cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear staining separately from extranuclear signal, because the reported tissue pattern is generally nuclear and the enhanced subcellular location is nucleoplasmic (HPA: tissue IHC and subcellular profile). An H-score combines the percentages of cells at 0–3+ intensity into a 0–300 score; report the percent positive alongside it (standard IHC practice). For spatial comparisons, count positive nuclei per mm² and normalise to total evaluable nuclei or the area of the same annotated cell compartment (standard IHC practice). Keep thresholds, counterstain, imaging settings and exclusion rules consistent, and report cell-type-specific results because staining levels differ among listed tissues and cells (standard IHC practice; HPA: tissue IHC).
When is a CBFA2T3-positive IHC result biologically credible?
A credible result shows reproducible staining in intact nuclei and a defined cell population, consistent with general nuclear expression and enhanced nucleoplasmic localisation (HPA: tissue IHC and subcellular profile). Check whether the stained cells fit the sampled tissue: reported high examples include caudate glial cells and cerebellar Purkinje cells (HPA: tissue IHC). Treat isolated edge staining, necrotic regions, diffuse staining without nuclear definition, or colour retained in a no-primary control as possible artefacts; investigate endogenous peroxidase when using DAB (standard IHC practice). The HPA tissue profile is Approved but reports low staining–RNA consistency, so corroborate unexpected patterns with controls or an independent antibody before drawing expression conclusions (HPA: reliability description; standard IHC practice).
Boster reagents
Best CBFA2T3 / Transcriptional corepressor CBFA2T3 IHC Antibodies
For CBFA2T3, A30478 has a human brain paraffin-section IHC image (catalog: A30478 IHC image caption), lists IF/ICC applications (catalog: A30478 applications), and states human and mouse reactivity (catalog: A30478 reactivity).
Real IHC data
Anti-MTG16 Antibody
Cat # A30478
A30478 will render with a paraffin-embedded human brain IHC image and a peptide-blocked comparison (catalog: A30478 IHC image caption). IF and ICC are listed applications, but no IF image is supplied (catalog: A30478 applications and IF image alts).
Which to pick: Choose A30478 for paraffin-section IHC; its own image caption shows human brain tissue and a peptide-blocked comparison, but does not report the fixative (catalog: A30478 IHC image caption). For IF/ICC, A30478 is a rabbit polyclonal antibody with both applications listed, although no IF image is supplied (catalog: A30478 host, dilution_raw, applications, and IF image alts). For human and mouse samples, A30478 lists reactivity with both species; the supplied IHC image shows human tissue only (catalog: A30478 reactivity and IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O75081 (MTG16_HUMAN, Transcriptional corepressor CBFA2T3).
- Human Protein Atlas. CBFA2T3 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CBFA2T3 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. CBFA2T3 antibody validation summary (3 antibodies).
- miR-27a and miR-27a* contribute to metastatic properties of osteosarcoma cells. Oncotarget 2015 — PMC4467124.
- Colony-Stimulating Factor 1 Receptor (CSF1R) Activates AKT/mTOR Signaling and Promotes T-Cell Lymphoma Viability. Clinical cancer research : an official journal of the American Association for Cancer Research 2020 — PMC7002219.
- Failure of human rhombic lip differentiation underlies medulloblastoma formation. Nature 2022 — PMC10026724.
- The Transcription Co-Repressors MTG8 and MTG16 Regulate Exit of Intestinal Stem Cells From Their Niche and Differentiation Into Enterocyte vs Secretory Lineages. Gastroenterology 2020 — PMC7607384.
- PubMed PMID:9596646 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:9790752 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CBFA2T3 (Transcriptional corepressor CBFA2T3). Expected localisation General nuclear 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: A30478 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A30478); 2. Fixation: Image fixative and duration unreported (datasheet A30478); 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-CBFA2T3, 1:100 - 1:300 (datasheet A30478); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep formalin fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining may diverge from tissue RNA levels (HPA tissue IHC); Isoform / epitope: Four isoforms; verify epitope coverage across variants (UniProt); Regulation: Brain, lymphoid tissue and pancreas have enhanced RNA (HPA tissue RNA). Evidence: UniProt O75081, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet A30478, PMC4467124, PMC7002219. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CBFA2T3/immunohistochemistry-ihc-guide.