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TRIM24 / Transcription intermediary factor 1-alpha · IHC design guide
Design Immunohistochemistry for TRIM24
Plan chromogenic TRIM24 IHC on paraffin sections using 2 μg/mL catalog antibody, as shown in its tissue captions (datasheet M03258-2). Compare staining with high thyroid glandular cell signal and undetected adipocyte signal, and score nuclear and cytoplasmic staining separately (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TRIM24 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, with cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Staining pattern | Cells across tissues: nuclear, sometimes cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03258-2) | |
| Positive control | Thyroid gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03258-2) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Early DNA damage lowers TRIM24 (UniProt) | |
| Isoform / epitope | 2 isoforms, Long and Short; check epitope coverage (UniProt) |
Section 1
Recommended TRIM24 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: M03258-2) with three published TRIM24 IHC protocols (PMC5076458; PMC8805319; PMC3661592).
| Sample | Paraffin-embedded human adenocarcinoma of the colon tissue; fixative not specified (datasheet M03258-2) |
| Fixation | Image fixative and duration unreported (datasheet M03258-2); 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 M03258-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03258-2) |
| Primary antibody | Mouse monoclonal (clone 4G6C2) anti-TRIM24, 2 μg/ml (datasheet M03258-2) |
| Primary incubation | Overnight at 4 °C (datasheet M03258-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03258-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TRIM24-positive staining in glandular cells of thyroid gland (HPA tissue IHC: High). HPA tissue profile: General nuclear expression with additional cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet: M03258-2); the published protocols provide citrate-based alternatives (PMC5076458; PMC8805319; PMC3661592).
Section 2
What Is the Expected TRIM24 Staining Pattern?
TRIM24 is predominantly nuclear and has no transmembrane segment (UniProt O15164: subcellular location; topology). In tissue IHC, expect nuclear staining in thyroid glandular cells at high intensity and in several other epithelial and trophoblastic cell populations at medium intensity (HPA: tissue IHC). HPA describes general nuclear expression with additional cytoplasmic staining in several tissues; its tissue IHC reliability is Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC).
| Distinct nuclear staining in thyroid glandular cells, with weaker staining in some other listed positive cell populations. | This fits the reported distribution: thyroid glandular cells are High; colon and duodenum glandular cells, esophageal squamous epithelial cells, ovarian follicle cells, and placental trophoblastic cells are Medium (HPA: tissue IHC). Compare cells within the same section before treating every difference in intensity as a technical failure (general IHC practice). |
| Predominantly cytoplasmic staining with little or no nuclear signal in an expected positive cell population. | Investigate the compartment call and possible staining artefact: predominant nuclear localisation is expected (UniProt O15164: subcellular location), although HPA reports additional cytoplasmic expression in several tissues (HPA: tissue IHC). Cytoplasmic signal alone cannot establish a false positive, because UniProt also records cytoplasmic localisation and context dependent movement to mitochondria (UniProt O15164: subcellular location). |
| Strong staining in a cell population listed as not detected, such as adipocytes or heart muscle cardiomyocytes. | Treat the unexpected result as a specificity question, including possible antibody cross-reactivity or endogenous detection activity (general IHC practice). HPA reports TRIM24 as Not detected in those particular cell populations (HPA: tissue IHC); that reference observation does not prove every specimen of that tissue must be negative. |
| Diffuse color across nuclei, cytoplasm, stroma, or empty areas without clear cell boundaries. | A uniform haze cannot be scored as the expected cell and compartment pattern (general IHC practice; HPA: general nuclear expression). Consider nonspecific binding, incomplete blocking, excess detection reagent, or endogenous detection activity as general IHC causes. Judge the signal against a matched negative detection control and a known positive tissue section (general IHC practice). |
| No discernible nuclear signal in thyroid glandular cells on a section expected to serve as a positive reference. | HPA reports High staining in thyroid glandular cells (HPA: tissue IHC), so absence of signal calls for a workflow check. Confirm that glandular cells are present and assess retrieval, antibody incubation, detection reagents, and counterstain with appropriate controls (general IHC practice). HPA's Approved rating has medium consistency with RNA data, so interpret a negative specimen cautiously (HPA: tissue IHC). |
💡Expected TRIM24 appearanceCall a result positive when expected cells show clear, predominantly nuclear chromogenic staining—High in thyroid glandular cells or Medium in listed positive populations (HPA: tissue IHC; UniProt O15164: subcellular location); diffuse background or staining that obscures cell boundaries is not a specific positive pattern (general IHC practice).
| Reference tissue and cell population | Choose the actual cell population named in the reference: thyroid glandular cells are High, whereas adipocytes and heart muscle cardiomyocytes are Not detected (HPA: tissue IHC). HPA also reports low tissue RNA specificity, so its staining list should guide comparison without becoming an absolute rule for every specimen (HPA: tissue IHC). |
| Reported IHC antibody validation | HPA043495 is listed as IHC Approved; HPA061717 has no IHC validation status in the supplied antibody record (HPA: antibody validation). The overall tissue IHC profile is Approved with medium consistency between staining and RNA expression (HPA: tissue IHC). Record which antibody produced the slide before comparing it with that reference pattern (general IHC practice). |
| Compartment and topology | Nuclear staining has the strongest routine expectation: UniProt calls TRIM24 predominantly nuclear, and HPA reports general nuclear tissue expression (UniProt O15164: subcellular location; HPA: tissue IHC). The protein has no transmembrane segment (UniProt O15164: topology). Additional cytoplasmic staining is documented, so assess its cell context alongside nuclear signal (HPA: tissue IHC). |
| Isoforms and epitope coverage | UniProt lists 2 isoforms, Long and Short (UniProt O15164: isoforms). The payload gives no antibody epitope or isoform coverage, so isoform specific staining cannot be predicted from it. If comparing antibodies, obtain their documented epitope information before attributing a difference in tissue staining to an isoform (general IHC practice). |
| IF/ICC Q: what localisation should an independent image show? | A: HPA's ICC-IF summary places TRIM24 in the nucleoplasm, with an enhanced main location; images are listed for A-431, U-251MG, U2OS, and Hep-G2 (HPA: subcellular ICC-IF). This supports a nuclear localisation check but does not supply an IHC staining intensity for those cell lines (HPA: subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| Thyroid positive reference is blank. | The expected glandular cells may be absent from the evaluated area, or an IHC workflow step may have failed (general IHC practice); thyroid glandular cells are High in the HPA reference (HPA: tissue IHC). | First identify intact glandular cells, then check retrieval, primary antibody incubation, chromogenic detection, and the positive control run (general IHC practice). Do not infer target specific fixation sensitivity from this result; none is supplied. |
| Signal is mainly cytoplasmic in a reference positive tissue. | The section may have background or a compartment assignment problem (general IHC practice). Additional cytoplasmic staining occurs in several tissues, but TRIM24 is predominantly nuclear (HPA: tissue IHC; UniProt O15164: subcellular location). | Inspect nuclear boundaries and compare the staining with a negative detection control and a known positive section (general IHC practice). Record the cytoplasmic component separately; do not score cytoplasmic color alone as the expected nuclear result. |
| A listed negative population stains strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice). HPA reports adipocytes and heart muscle cardiomyocytes as Not detected (HPA: tissue IHC). | Check a control omitting the primary antibody for detection related signal, then compare another documented reference cell population and the antibody's IHC validation status (general IHC practice; HPA: antibody validation). |
| The whole section shows diffuse brown haze. | Nonspecific binding, inadequate blocking, or excessive detection signal are general IHC possibilities (general IHC practice). A haze lacks the predominantly nuclear pattern reported for TRIM24 (UniProt O15164: subcellular location; HPA: tissue IHC). | Review blocking and detection controls, then adjust reagent conditions using the antibody's documented IHC instructions (general IHC practice). Score only staining that can be assigned to cells and compartments. |
| A faint section is being called negative. | Some named populations are Low, including lung alveolar cells and salivary gland glandular cells (HPA: tissue IHC). Weak target signal can be hard to separate from counterstain or background (general IHC practice). | Compare the same cell population with a positive reference and a negative detection control, and inspect nuclei before assigning an intensity category (general IHC practice). Avoid using a low staining population as the sole assay check (HPA: tissue IHC). |
| Results differ between two antibodies or between IHC and IF/ICC. | Antibody validation and assay context differ: HPA043495 is IHC Approved, while the supplied record gives HPA061717 no IHC status; HPA's ICC-IF localisation is nucleoplasmic (HPA: antibody validation; HPA: subcellular ICC-IF). | Keep each antibody and assay result identified, compare nuclear localisation first, and review documented epitope coverage before proposing an isoform explanation (general IHC practice; UniProt O15164: 2 isoforms). |
Experiment evidence
IHC & IF images showing TRIM24 staining
M03258-2 · IHCIHC analysis of TRIM24 using anti-TRIM24 antibody (M03258-2). TRIM24 was detected in a paraffin-embedded section of human adenocarcinoma of
M03258-2 · IHCIHC analysis of TRIM24 using anti-TRIM24 antibody (M03258-2). TRIM24 was detected in a paraffin-embedded section of human thyroid cancer tis
M03258-2 · IFIF analysis of TRIM24 using anti-TRIM24 antibody (M03258-2). TRIM24 was detected in an immunocytochemical section of MCF-7 cells. Enzyme ant
A03258 · IFImmunocytochemistry of TRIM24 in EL4 cells with TRIM24 antibody at 2.5 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000122779-TRIM24/tissue/cerebral+cortex#imid_11916695
HPA · Thyroid glandHuman Protein Atlas IHC, Thyroid gland. Source: https://www.proteinatlas.org/ENSG00000122779-TRIM24/tissue/thyroid+gland#imid_11916639
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000122779-TRIM24/tissue/colon#imid_11916605
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000122779-TRIM24/tissue/liver#imid_11916622
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043495; green = TRIM24, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA043495; green = TRIM24, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TRIM24 IHC & IF
🧪Run thyroid gland first and assess staining in its glandular cells (HPA: High in thyroid glandular cells). Use adipose tissue as the negative comparator, assessing adipocytes (HPA: Not detected in adipocytes); on the thyroid slide, cells used as internal negatives should lack specific nuclear staining, but HPA does not identify a particular thyroid cell type as negative (HPA: thyroid glandular cells High).
Positive control tissue: Thyroid 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 TRIM24 in A-431, U-251MG, U2OS, Hep-G2, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a matched mouse IgG isotype control, and a validated TRIM24 knockout specimen as a biological negative (caption: mouse anti-TRIM24 antibody; standard IHC controls). Block endogenous peroxidase for HRP/DAB detection and distinguish staining of thyroid glandular cell nuclei from background in the colloid (caption: HRP/DAB; HPA: thyroid glandular cells High).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU caption does not state the fixative (M03258-2 caption: fixative not stated). The documented paraffin-section method uses heat-mediated EDTA retrieval at pH 8.0; the supplied evidence does not establish that frozen sections or IF/ICC are easier (M03258-2 caption: EDTA pH 8.0 retrieval; HPA subcellular: nucleoplasm). In thyroid sections, assess nuclear staining separately from colloid background (HPA: thyroid glandular cells High; UniProt O15164: predominantly nuclear).
HPA tissue IHC evidence for TRIM24
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Endothelial cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TRIM24 IHC Tips
Troubleshoot TRIM24 staining in paraffin sections by checking retrieval, nuclear localisation and cell type before comparing staining intensity across samples.
What retrieval should I use when TRIM24 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet M03258-2). This is the retrieval used for the catalog antibody in a paraffin section of human colon adenocarcinoma, followed by 2 μg/ml primary antibody overnight at 4°C (datasheet M03258-2). If staining remains weak, compare sections processed with the same heat exposure and check that the primary incubation and detection reagents performed consistently (standard IHC practice). Judge improvement in nuclei first, because TRIM24 is predominantly nuclear (UniProt O15164) and HPA reports general nuclear tissue staining (HPA tissue IHC). Record retrieval conditions for each run so intensity differences are not mistaken for biological differences (standard IHC practice).
Could fixation explain weak or uneven TRIM24 staining?
The selected paraffin-section caption does not report a fixative, so target-specific fixation sensitivity is unknown (datasheet M03258-2). Do not infer a preferred fixative or fixation time from TRIM24’s intracellular localisation or modified residues (UniProt O15164). For a troubleshooting comparison, process adjacent sections with the same EDTA retrieval at pH 8.0 and the same 2 μg/ml primary incubation overnight at 4°C (datasheet M03258-2). Document the available fixation history and compare well-preserved areas before attributing uneven signal to antigen loss (standard IHC practice). Exclude damaged or detached regions from scoring because tissue integrity affects chromogenic interpretation (standard IHC practice).
How should I assess cytoplasmic staining when I expect nuclear TRIM24?
Score nuclear and cytoplasmic DAB separately: TRIM24 is predominantly nuclear, while cytoplasmic and mitochondrial localisation are also recorded (UniProt O15164). HPA describes general nuclear expression with additional cytoplasmic expression in several tissues, so cytoplasmic colour alone should not establish positivity (HPA tissue IHC). Compare intact cells within the same section for a reproducible compartment pattern and inspect the hematoxylin counterstain to locate nuclei (standard IHC practice). In the selected colon adenocarcinoma section, the documented assay used 2 μg/ml primary antibody and DAB detection (datasheet M03258-2). Report the fraction of stained nuclei separately from cytoplasmic intensity when the two disagree (standard IHC practice).
Could isoforms or epitope accessibility account for inconsistent TRIM24 staining?
TRIM24 has 2 annotated isoforms, Long and Short, but the supplied product caption does not identify its antibody epitope or establish isoform coverage (UniProt O15164; datasheet M03258-2). Its bromodomain spans residues 899–1004, and the record lists multiple modified residues, so an epitope assignment would require antibody-specific documentation (UniProt O15164). First compare staining after the documented EDTA retrieval at pH 8.0, keeping section processing and primary incubation consistent (datasheet M03258-2; standard IHC practice). If a second antibody with a documented, distinct epitope is available, concordant nuclear patterns can help assess specificity (standard IHC practice). Do not label discordant staining as an isoform difference without epitope and isoform evidence (standard IHC practice).
How can IF help assess TRIM24 localisation alongside the IHC result?
Use IF as a separate localisation check, pairing TRIM24 with a marker for the expected cell type and a nuclear counterstain (standard IF practice). HPA reports enhanced nucleoplasmic localisation in ICC/IF images and lists A-431, U-251MG, U2OS and Hep-G2 among the imaged cell lines (HPA subcellular). Choose a fluorophore channel away from prominent specimen autofluorescence and inspect single-channel controls before interpreting overlap (standard IF practice). Because TRIM24 is intracellular and has no transmembrane segment, permeabilisation must allow antibody access to the nucleus; optimise it for the IF specimen and antibody (UniProt O15164; standard IF practice). The pH 8.0 EDTA retrieval and 2 μg/ml primary concentration describe the selected paraffin-section IHC assay, not an IF protocol (datasheet M03258-2).
What should I check when DAB obscures nuclear TRIM24 staining?
Check a no-primary control for signal from the detection system and inspect tissue edges, folds and damaged areas for uneven DAB deposition (standard IHC practice). The selected assay used a goat serum block at 10%, a peroxidase-conjugated secondary and DAB chromogen (datasheet M03258-2). An endogenous peroxidase block is a general chromogenic IHC step; the selected caption does not state whether one was used (standard IHC practice; datasheet M03258-2). Compare background with the documented 2 μg/ml primary incubation before changing antibody concentration (datasheet M03258-2). Interpret diffuse staining against the expected predominantly nuclear pattern and retain only clearly localised signal for scoring (UniProt O15164; standard IHC practice).
How should I quantify TRIM24 across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear TRIM24 as the percentage of positive cells plus intensity, or combine those measures in an H-score (standard IHC practice). Count a consistent number of evaluable cells or fields per section and normalise positive counts to total cells of the same type (standard IHC practice). If reporting positive-cell density, divide by analysed viable tissue area in mm² and exclude folds, edges and necrosis (standard IHC practice). Record cytoplasmic staining separately because TRIM24 is predominantly nuclear but HPA also reports cytoplasmic tissue staining (UniProt O15164; HPA tissue IHC). Hold retrieval at EDTA pH 8.0 and detection conditions constant when comparing the catalog assay across sections (datasheet M03258-2; standard IHC practice).
How can I distinguish genuine TRIM24 staining from section artefacts?
A convincing result has cell-associated staining in an interpretable compartment, especially nuclei, consistent with TRIM24’s predominant nuclear localisation (UniProt O15164). HPA reports high staining in thyroid glandular cells and medium staining in colon glandular cells, while also describing additional cytoplasmic expression in several tissues (HPA tissue IHC). Treat signal confined to a cut edge, fold or necrotic area as suspect, and compare it with intact cells and a no-primary control (standard IHC practice). Investigate diffuse DAB in the control as possible endogenous enzyme or detection background before scoring target signal (standard IHC practice). HPA rates its tissue staining Approved with medium consistency against RNA expression, so report compartment, cell type and controls alongside intensity (HPA tissue IHC; standard IHC practice).
Boster reagents
Best TRIM24 / Transcription intermediary factor 1-alpha IHC Antibodies
The catalog shows real TRIM24 IHC data from human paraffin sections (M03258-2 IHC captions) and IF/ICC data from MCF-7 and EL4 cells (M03258-2 and A03258 IF captions).
Real IHC data
Anti-TRIM24 Antibody ® (monoclonal, 4G6C2)
Cat # M03258-2
Real IF data
Anti-TRIM24 Antibody
Cat # A03258
M03258-2 has IHC images from human paraffin sections of colon adenocarcinoma, thyroid cancer, ovarian serous tumor, and breast cancer, plus IF in MCF-7 cells (M03258-2 image captions). A03258 has ICC and IF images in EL4 cells, with mouse ICC/IF validation stated in its catalog entry (A03258 image captions and validation text).
Which to pick: Choose M03258-2 for tissue IHC: its own caption documents a human paraffin section with EDTA retrieval, while the fixative is unreported (M03258-2 IHC caption). For IF/ICC, choose M03258-2 for MCF-7 cells or A03258 for EL4 cells (respective IF captions). A03258 lists human, mouse, and rat reactivity for cross-species planning, but its pictured IF/ICC validation is in mouse EL4 cells (A03258 catalog and validation text).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O15164 (TIF1A_HUMAN, Transcription intermediary factor 1-alpha).
- Human Protein Atlas. TRIM24 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TRIM24 subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. TRIM24 antibody validation summary (2 antibodies).
- Clinical significance and prognostic value of TRIM24 expression in esophageal squamous cell carcinoma. Aging 2016 — PMC5076458.
- Trim24 prompts tumor progression via inducing EMT in renal cell carcinoma. Open medicine (Warsaw, Poland) 2020 — PMC7718642.
- High expression of TRIM24 predicts worse prognosis and promotes proliferation and metastasis of epithelial ovarian cancer. Journal of ovarian research 2022 — PMC8805319.
- TRIM24 overexpression is common in locally advanced head and neck squamous cell carcinoma and correlates with aggressive malignant phenotypes. PloS one 2013 — PMC3661592.
- PubMed PMID:9115274 — UniProt-cited evidence.
- PubMed PMID:10022127 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TRIM24 (Transcription intermediary factor 1-alpha). Expected localisation Nuclear, with cytoplasmic staining in several tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03258-2). Validated IHC antibody: M03258-2 at 2 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Thyroid gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human adenocarcinoma of the colon tissue; fixative not specified (datasheet M03258-2); 2. Fixation: Image fixative and duration unreported (datasheet M03258-2); 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 M03258-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03258-2); 8. Primary antibody: Mouse monoclonal (clone 4G6C2) anti-TRIM24, 2 μg/ml (datasheet M03258-2); 9. Primary incubation: Overnight at 4 °C (datasheet M03258-2). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03258-2); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 2 isoforms, Long and Short; check epitope coverage (UniProt); Regulation: Early DNA damage lowers TRIM24 (UniProt). Evidence: UniProt O15164, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm., Datasheet M03258-2, PMC5076458, PMC8805319. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TRIM24/immunohistochemistry-ihc-guide.