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TAF15 / TATA-binding protein-associated factor 2N · IHC design guide
Design Immunohistochemistry for TAF15
Plan TAF15 staining in paraffin sections using the widespread nuclear pattern reported in tissue (HPA tissue IHC). The guide covers cell-type controls, scoring, and the catalog antibody’s 1:50 starting dilution (datasheet M03567-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TAF15 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 staining in tissue cells (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across diverse cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03567-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes and ovarian stromal cells may lack signal (HPA tissue IHC) | |
| Regulation | Broad expression across fetal and adult tissues (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended TAF15 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: M03567-1) is followed by published TAF15 IHC methods for GIST, human organ arrays, and cholangiocarcinoma tissue (PMC10237090; PMC2478660; PMC12615741).
| Sample | Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M03567-1) |
| Fixation | Image fixative and duration unreported (datasheet M03567-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M03567-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03567-1) |
| Primary antibody | Rabbit monoclonal (clone 27T75) anti-TAF15, 1:50 (datasheet M03567-1) |
| Primary incubation | Overnight at 4 °C (datasheet M03567-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03567-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TAF15-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: M03567-1). Microwave citrate retrieval at pH 6.0 is a published alternative (PMC2478660).
Section 2
What Is the Expected TAF15 Staining Pattern?
TAF15 staining should be predominantly nuclear across many cell types, consistent with HPA's ubiquitous nuclear IHC profile and UniProt's nuclear localisation (HPA: Supported, with medium consistency between staining and RNA data; UniProt Q92804: nucleus). High staining is reported in selected glandular, hematopoietic, glial, neuronal and endothelial cells (HPA: tissue IHC). Cytoplasmic signal can occur because TAF15 shuttles between compartments; it has no transmembrane segment (UniProt Q92804: localisation and topology).
| Clear nuclear staining in adrenal glandular or bone marrow hematopoietic cells. | This matches reported high staining in those cells (HPA: High in adrenal glandular and bone marrow hematopoietic cells). Judge the pattern within the identified cells, with the nuclear signal as the main positive finding (HPA: ubiquitous nuclear expression). |
| Predominantly membrane staining, or strong cytoplasmic staining with little nuclear signal. | Membrane staining conflicts with TAF15's reported location and lack of a transmembrane segment (UniProt Q92804: localisation and topology). Cytoplasmic signal alone is less decisive because TAF15 shuttles; check a known-positive nuclear pattern before calling the result specific (UniProt Q92804: shuttling; HPA: nuclear IHC). |
| Strong staining in adipocytes or ovarian stroma cells, especially without nearby nuclear staining. | These cell types are reported as not detected, so unexpected staining warrants a specificity and detection-control review (HPA: adipocytes and ovarian stroma cells Not detected; general IHC practice). Their HPA result is a cell-level observation, not proof that every sample must be negative (HPA: tissue IHC). |
| Diffuse colour across nuclei, cytoplasm and tissue spaces. | A pattern without cell boundaries or nuclear enrichment is difficult to assign to TAF15 (HPA: ubiquitous nuclear expression). Review the no-primary control and blocking, washing and detection steps for background or endogenous activity (general chromogenic IHC practice). |
| No nuclear signal in a section containing a reported high-staining cell population. | First confirm that the expected cell population is present (HPA: High in the specified cells). Then check antibody and detection controls and the applied IHC-P conditions; an absent signal in one section does not overturn the supported, medium-consistency HPA pattern (HPA: tissue IHC reliability; general IHC practice). |
💡Expected TAF15 appearanceCall positive when the identified cells show predominantly nuclear chromogenic signal, potentially high in HPA-listed high-staining populations; isolated membrane staining or diffuse, unstructured colour is suspect (HPA: ubiquitous nuclear expression and tissue IHC; UniProt Q92804: topology; general IHC practice).
| Cell identity and tissue context | TAF15 is described as ubiquitous, yet HPA reports high staining in some cell populations, low staining in cardiomyocytes and splenic red-pulp cells, and no detection in adipocytes or ovarian stroma cells (UniProt Q92804: tissue specificity; HPA: tissue IHC). Score identified cell types rather than treating an entire tissue as uniformly positive. |
| Compartment and topology | HPA describes a nuclear tissue-IHC profile; UniProt places TAF15 in the nucleus and cytoplasm and reports no transmembrane segment (HPA: tissue IHC; UniProt Q92804: localisation and topology). Use nuclear enrichment as the main IHC readout, while interpreting cytoplasmic signal in context. |
| Antibody evidence | The supplied HPA antibody record lists HPA052059 as Supported for IHC and Enhanced for ICC; HPA063647 has no supplied IHC status (HPA: antibody validation). These labels support application-specific interpretation, but do not establish that an unlisted catalog antibody has the same performance. |
| Isoforms and modifications | UniProt lists Long and Short isoforms and multiple modified residues, including methylation, phosphorylation and acetylation (UniProt Q92804: isoforms and modified residues). No epitope or target-specific fixation-effect evidence is supplied, so these entries do not predict a change in IHC signal. |
| IF/ICC Q&A: What localisation is expected? | Predominantly nucleoplasmic signal is expected in the reported ICC-IF images (HPA: Nucleoplasm, enhanced). This is an IF/ICC localisation reference, not an IHC-P protocol or evidence that the same antibody conditions apply to paraffin sections (HPA: subcellular and antibody records). |
| Situation | Likely cause | Next action |
|---|---|---|
| Weak or absent nuclear staining in a known high-staining cell population. | The expected population may be missing from the section, or the IHC detection run may have failed (HPA: cell-specific High staining; general IHC practice). | Identify the relevant cells, then compare a known-positive section and detection controls run under the same IHC-P conditions (HPA: tissue IHC; general IHC practice). |
| Strong signal is confined to membranes. | That distribution disagrees with the reported nuclear profile and lack of a transmembrane segment (HPA: nuclear tissue IHC; UniProt Q92804: topology). | Inspect the no-primary control and reassess staining with an IHC-validated antibody before assigning membrane signal to TAF15 (general IHC practice; HPA: antibody validation). |
| Cytoplasmic signal dominates and nuclei appear negative. | TAF15 can shuttle to cytoplasm, but the supplied tissue-IHC profile is nuclear (UniProt Q92804: shuttling; HPA: ubiquitous nuclear expression). | Compare cell morphology and a known-positive nuclear control; report cytoplasmic predominance as an observation pending corroboration (HPA: tissue IHC; general IHC practice). |
| Diffuse background obscures the cells. | Nonspecific binding, inadequate washing or endogenous chromogenic-detection activity can produce background (general IHC practice). | Review the no-primary control, blocking and wash steps, and the appropriate endogenous-activity control for the chosen detection system (general IHC practice). |
| Adipocytes or ovarian stroma cells appear strongly positive. | The observation differs from HPA's Not detected result for those cell types; cross-reactivity or detection background is possible (HPA: tissue IHC; general IHC practice). | Verify cell identity and nuclear localisation, then compare no-primary and known-positive controls before interpreting the signal (HPA: tissue IHC; general IHC practice). |
| An IF/ICC image looks nucleoplasmic, but the paraffin-section IHC result is unclear. | HPA's nucleoplasmic ICC-IF localisation and its tissue-IHC observations are distinct application records (HPA: subcellular and tissue IHC). | Use the ICC-IF result to assess localisation only; resolve the IHC result with section-level controls and an antibody validated for IHC (HPA: antibody validation; general IHC practice). |
Experiment evidence
IHC & IF images showing TAF15 staining
M03567-1 · IHCIHC analysis of TAF15 using anti-TAF15 antibody (M03567-1). TAF15 was detected in a paraffin-embedded section of human colorectal adenocarci
M03567-1 · IHCIHC analysis of TAF15 using anti-TAF15 antibody (M03567-1). TAF15 was detected in a paraffin-embedded section of human liver cancer tissue.
M03567-1 · IFIF analysis of TAF15 using anti-TAF15 antibody (M03567-1) and anti-Beta Tubulin antibody (M01857-3). TAF15 was detected in immunocytochemica
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000270647-TAF15/tissue/cerebral+cortex#imid_14336756
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000270647-TAF15/tissue/colon#imid_14330850
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000270647-TAF15/tissue/liver#imid_14330843
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000270647-TAF15/tissue/kidney#imid_14330844
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052059; green = TAF15, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052059; green = TAF15, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TAF15 IHC & IF
🧪Run bone marrow first: hematopoietic cells should stain (HPA: High in bone marrow hematopoietic cells). Use adipose tissue as the comparison tissue; its adipocytes are listed as Not detected (HPA: Not detected in adipocytes). No confirmed negative cell type is identified within the bone marrow slide, so cells without specific signal should show counterstain only, at the background level of the no-primary control (HPA: bone marrow row; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TAF15 in HeLa, MCF-7, U2OS, A-549, SiHa, U2OS, siRNA 1 (10x), U2OS, siRNA 2 (10x), U2OS, scrambled (10x), NIH 3T3, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a rabbit IgG control matched to the primary antibody’s isotype; use TAF15 knockout material as a biological specificity control where available (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase before chromogenic detection, especially in bone marrow, where hematopoietic cells can contribute peroxidase background (HPA: bone marrow hematopoietic cells; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state the fixative (caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 and primary incubation at 1:50 overnight at 4°C; it does not establish that retrieval is required under other conditions (caption: M03567-1 tissue IHC). Whether frozen sections or IF are easier is unreported, although ICC-IF images support assessment of nucleoplasmic localization; account for endogenous peroxidase background when scoring bone marrow (HPA: nucleoplasm, enhanced; HPA: bone marrow hematopoietic cells; standard IHC practice).
HPA tissue IHC evidence for TAF15
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Caudate | Glial cells | High | Protein (IHC) | HPA → |
| Cerebellum | Bergmann glia - nucleus | High | Protein (IHC) | HPA → |
Section 3
Advanced TAF15 IHC Tips
Troubleshoot TAF15 staining in paraffin sections using the catalog antibody’s tissue IHC conditions, expected nuclear pattern, and matched controls.
Which retrieval conditions should I try first for weak TAF15 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet M03567-1). The selected paraffin-section image used that retrieval, followed by 10% goat serum blocking and antibody at 1:50 overnight at 4°C (datasheet M03567-1). If staining is weak, check section adhesion, heating consistency, antibody dilution, and detection before testing another retrieval buffer as a fallback (standard IHC practice). Compare any change on adjacent sections with the same detection conditions, using nuclear staining in an expected positive cell population as the readout (HPA tissue IHC: ubiquitous nuclear expression).
How should I troubleshoot fixation when TAF15 nuclear staining is weak?
The selected image describes a paraffin-embedded section but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet M03567-1). Record the actual fixative and fixation duration for each specimen, then compare sections processed with the same retrieval and antibody conditions (standard IHC practice). Examine morphology and staining across the section: uneven preservation can complicate assessment of a nuclear signal (standard IHC practice; HPA tissue IHC: ubiquitous nuclear expression). Use a known positive section in the same run to separate a specimen-processing problem from a broader staining failure (standard IHC practice).
Should cytoplasmic TAF15 staining be scored as positive in paraffin sections?
Prioritize a reproducible nuclear signal when assessing tissue sections, because the tissue IHC profile reports ubiquitous nuclear expression (HPA tissue IHC). More specifically, subcellular imaging places TAF15 in the nucleoplasm (HPA subcellular: nucleoplasm). Cytoplasmic signal warrants careful review because TAF15 can shuttle between nucleus and cytoplasm, but that annotation alone does not validate a diffuse cytoplasmic IHC pattern (UniProt Q92804: subcellular location). Compare the signal with matched controls and nuclear counterstain, and record nuclear and cytoplasmic staining separately rather than combining them in one score (standard IHC practice).
Could isoforms or epitope accessibility explain inconsistent TAF15 staining?
TAF15 has 2 annotated isoforms, Long and Short, but the supplied antibody evidence does not map its epitope or establish which isoforms it detects (UniProt Q92804: isoforms; datasheet M03567-1). Its RNA recognition motif spans residues 234–320, and annotated modifications include methylation, phosphorylation, and acetylation (UniProt Q92804: domains and modified residues). Those annotations identify possibilities for follow-up; they do not show that any particular modification changes this antibody’s staining. Compare adjacent sections under identical processing conditions and seek epitope mapping or an independent antibody before attributing a staining difference to isoform expression (standard IHC practice).
How can IF help assess the TAF15 pattern seen by chromogenic IHC?
On the separate IF/ICC guide, assess TAF15 with a validated glial marker in a suitable specimen: glial cells in the caudate have high reported tissue staining (HPA tissue IHC: caudate glial cells). Choose a far-red TAF15 fluorophore if the specimen shows strong short-wavelength autofluorescence, and include single-label controls for multiplex interpretation (standard IF practice). Because TAF15 is intracellular, lacks a transmembrane segment, and is enriched in the nucleoplasm, assess permeabilisation sufficient for nuclear antibody access (UniProt Q92804: topology and location; HPA subcellular: nucleoplasm). Compare compartment patterns with the chromogenic result, while treating fixation and antibody performance in IF as separate validation questions (standard IF practice).
What should I adjust if TAF15 staining is diffuse or obscured by background?
First inspect no-primary and secondary-only controls to locate background from the detection system (standard IHC practice). The selected image used 10% goat serum blocking, a peroxidase-conjugated secondary, and DAB development; those are documented conditions for that image, not proof they will remove background in every section (datasheet M03567-1). Include a peroxidase block and assess endogenous pigment or enzyme signal before interpreting brown deposits as TAF15 (standard chromogenic IHC practice). Recheck antibody concentration and washing against a positive section, and favor crisp nuclear signal over diffuse staining when judging specificity (HPA tissue IHC: ubiquitous nuclear expression; standard IHC practice).
How should I quantify TAF15 staining across tissue compartments? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, because reported TAF15 staining varies by cell type even within the tissue survey (HPA tissue IHC). Score the percentage of positive nuclei and nuclear intensity, then report an H-score if its intensity categories are defined in advance (standard IHC practice). Normalize counts or positive-cell density per mm² to viable tissue area and report the number of eligible cells, avoiding necrotic or damaged regions (standard IHC practice). Keep cytoplasmic staining as a separate measure and apply the same threshold, counterstain review, and image settings across comparison groups (UniProt Q92804: nuclear-cytoplasmic shuttling; standard IHC practice).
How can I distinguish genuine TAF15 staining from section artefacts?
A credible positive pattern is predominantly nuclear within intact cells, consistent with the tissue IHC profile and nucleoplasmic imaging (HPA tissue IHC; HPA subcellular: nucleoplasm). Check the identity of the stained cells: adrenal glandular cells and bone marrow hematopoietic cells are reported high, whereas adipocytes are reported not detected (HPA tissue IHC). Treat staining concentrated at section edges, in necrosis, or over damaged tissue as suspect until it persists in well-preserved regions (standard IHC practice). Use no-primary controls and peroxidase blocking to assess nonspecific DAB deposition or endogenous enzyme activity before calling a weak signal positive (standard chromogenic IHC practice).
Boster reagents
Best TAF15 / TATA-binding protein-associated factor 2N IHC Antibodies
M03567-1 has IHC images from human paraffin sections and an IF image from HeLa cells (catalog image captions). Human, Mouse, and Rat reactivity is listed (catalog reactivity).
Real IHC data
Anti-TAF15 Rabbit Monoclonal Antibody
Cat # M03567-1
M03567-1 has IHC images from paraffin sections of human colorectal adenocarcinoma, liver cancer, placenta, and prostate cancer (catalog IHC image captions). Its IF image shows HeLa cells, and its listed applications include IHC and ICC/IF (catalog IF image caption; catalog applications).
Which to pick: For tissue IHC, choose M03567-1: its IHC captions document EDTA retrieval at pH 8.0 and a 1:50 primary dilution in human paraffin sections; the fixative is unreported (catalog IHC image captions). For IF/ICC, the same rabbit monoclonal, clone 27T75, has a HeLa IF image using 1:50 primary dilution (catalog host/clone; catalog IF image caption). For mouse or rat work, M03567-1 lists both species as reactive and lists IHC and ICC/IF applications; the supplied images show human tissue and HeLa cells (catalog reactivity/applications; catalog image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q92804 (RBP56_HUMAN, TATA-binding protein-associated factor 2N).
- Human Protein Atlas. TAF15 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. TAF15 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. TAF15 antibody validation summary (2 antibodies).
- TATA-box-binding protein-associated factor 15 is a novel biomarker that promotes cell proliferation and migration in gastrointestinal stromal tumor. World journal of gastroenterology 2023 — PMC10237090.
- A repeat expansion in GOLGA8A is a major risk factor for atypical frontotemporal lobar degeneration with ubiquitin-positive inclusions. Nature genetics 2026 — PMC13083237.
- The multifunctional FUS, EWS and TAF15 proto-oncoproteins show cell type-specific expression patterns and involvement in cell spreading and stress response. BMC cell biology 2008 — PMC2478660.
- TAF15 in tumor-associated macrophages enhances protumorigenic polarization and promotes cholangiocarcinoma progression. JHEP reports : innovation in hepatology 2025 — PMC12615741.
- PubMed PMID:8954779 — UniProt-cited evidence.
- PubMed PMID:8890175 — UniProt-cited evidence.
- PubMed PMID:9795213 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TAF15 (TATA-binding protein-associated factor 2N). Expected localisation Nuclear staining in tissue cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03567-1). Validated IHC antibody: M03567-1 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colorectal adenocarcinoma tissue; fixative not specified (datasheet M03567-1); 2. Fixation: Image fixative and duration unreported (datasheet M03567-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet M03567-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03567-1); 8. Primary antibody: Rabbit monoclonal (clone 27T75) anti-TAF15, 1:50 (datasheet M03567-1); 9. Primary incubation: Overnight at 4 °C (datasheet M03567-1). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Adipocytes and ovarian stromal cells may lack signal (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Broad expression across fetal and adult tissues (UniProt). Evidence: UniProt Q92804, HPA tissue IHC · Supported, HPA ICC-IF · nucleoplasm., Datasheet M03567-1, PMC10237090, PMC2478660. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TAF15/immunohistochemistry-ihc-guide.