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TRIM25 / E3 ubiquitin/ISG15 ligase TRIM25 · IHC design guide
Design Immunohistochemistry for TRIM25
Plan paraffin-section TRIM25 IHC around its general cytoplasmic tissue pattern (HPA tissue IHC). The catalog antibody has an IHC starting dilution of 1:50 (datasheet: M03232); assess DAB background when using marrow sections (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TRIM25 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 cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining; high in marrow hematopoietic cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03232) | |
| Positive control | Bone marrow+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 M03232) | |
| Caveat | Endogenous peroxidase in marrow may mimic DAB staining (HPA tissue IHC; standard IHC practice) | |
| Regulation | Staining-linked regulation unreported (HPA tissue IHC) | |
| Isoform / epitope | No annotated isoforms or processing to alter the epitope map (UniProt) |
Section 1
Recommended TRIM25 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 retrieval (datasheet M03232). The published options below report TRIM25 IHC in liver and prostate cancer tissue (PMC13079618; PMC9408812; PMC5323126).
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet M03232) |
| Fixation | Image fixative and duration unreported (datasheet M03232); 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 M03232); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03232) |
| Primary antibody | Rabbit monoclonal (clone ADFG-20) anti-TRIM25, 1:50 (datasheet M03232) |
| Primary incubation | Overnight at 4 °C (datasheet M03232) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03232) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TRIM25-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet M03232); the emitted article excerpts do not specify retrieval conditions.
Section 2
What Is the Expected TRIM25 Staining Pattern?
TRIM25 should appear mainly in the cytoplasm on paraffin IHC, with staining in cell types such as breast glandular cells, bronchial respiratory epithelium, and bone marrow hematopoietic cells (HPA: general cytoplasmic expression; High in these cells). Nuclear localisation is also reported (UniProt Q14258: nucleus), though HPA rates its nucleoplasmic IF localisation uncertain. TRIM25 has no transmembrane segment (UniProt Q14258: topology). HPA rates its tissue IHC evidence Approved, with medium consistency against RNA expression (HPA: reliability).
| Cytoplasmic chromogen is clear in breast glandular cells or bronchial respiratory epithelium, with cell boundaries still readable (HPA: High in both cell types). | This matches the predominant tissue pattern (HPA: general cytoplasmic expression). Compare intensity within the identified cell population; the HPA High category describes reported staining, not a required score for every section (HPA: tissue IHC). |
| Signal is confined to cell membranes or extracellular material, without a convincing cytoplasmic component (HPA: general cytoplasmic expression; UniProt Q14258: no transmembrane segment). | That distribution does not fit the reported predominant location (HPA: general cytoplasmic expression). Check whether pigment, edge staining, or nonspecific detection reproduces it in controls (standard IHC practice). Do not reject all nuclear signal: UniProt also lists the nucleus (UniProt Q14258: subcellular location). |
| The strongest chromogen is in adipocytes or cardiomyocytes while the expected positive cells are weak (HPA: Not detected in adipocytes and cardiomyocytes). | This conflicts with the reported tissue pattern and raises cross-reactivity or endogenous detection activity as possibilities (HPA: tissue IHC; standard IHC practice). Identify the stained cell type before assigning a tissue-level score; staining elsewhere in the section does not establish staining in that cell type (standard IHC practice). |
| Color coats many structures evenly, including spaces between cells, and obscures cell boundaries (standard IHC practice). | Treat this as background until a cell-associated pattern survives appropriate negative controls (standard IHC practice). Broad expression alone cannot explain extracellular haze: HPA describes general cytoplasmic expression and low tissue specificity, not uniform deposition across a section (HPA: tissue IHC; RNA specificity). |
| No cytoplasmic signal appears in breast glandular cells or bronchial respiratory epithelium (HPA: High in both cell types). | First assess tissue preservation, the staining run, and a suitable positive control (standard IHC practice). An absent signal is discordant with these HPA observations, but one section cannot establish that TRIM25 is absent: HPA reports medium consistency between staining and RNA data (HPA: reliability description). |
💡Expected TRIM25 appearanceCall a result consistent when identifiable breast glandular or bronchial respiratory epithelial cells show clear, predominantly cytoplasmic chromogen, potentially strong relative to adjacent cells (HPA: High in these cell types; general cytoplasmic expression); membrane-only deposits or diffuse extracellular color warrant investigation as possible false positives (UniProt Q14258: no transmembrane segment; standard IHC practice).
| Compartment and readout (HPA: tissue IHC; UniProt Q14258: subcellular location) | Score cytoplasmic staining first because that is HPA's reported tissue pattern (HPA: general cytoplasmic expression). Record nuclear staining separately: UniProt lists the nucleus, while HPA's nucleoplasmic ICC-IF assignment is uncertain (UniProt Q14258: nucleus; HPA: nucleoplasm uncertain). |
| Choice of comparison cells (HPA: tissue IHC) | Breast glandular cells, bronchial respiratory epithelial cells, and bone marrow hematopoietic cells are reported High; adipocytes, cardiomyocytes, and skeletal myocytes are Not detected (HPA: cell-level tissue IHC). These are comparison patterns, not universal acceptance thresholds (HPA: Approved, medium consistency). |
| Strength of the IHC evidence (HPA: antibody and tissue validation) | The listed antibody HPA005909 is IHC Approved, while HPA describes medium consistency between tissue staining and RNA expression (HPA: HPA005909 IHC Approved; reliability description). Interpret unexpected cells with controls rather than treating an isolated stain as definitive target identification (standard IHC practice). |
| Topology and processing (UniProt Q14258: topology and processing) | TRIM25 has no transmembrane segment or signal peptide, and its annotated chain spans residues 1–630 (UniProt Q14258: topology; processing). A membrane-only or extracellular IHC pattern therefore needs independent scrutiny; these annotations do not predict fixation sensitivity (UniProt Q14258: topology; standard IHC practice). |
| What about IF/ICC? (HPA: subcellular; HPA005909 ICC Supported) | On its separate guide page, expect mainly cytosol, with nucleoplasm reported as uncertain and nuclear bodies as an additional uncertain location (HPA: ICC-IF subcellular). The listed antibody has ICC Supported status; that status does not establish an IHC nuclear scoring rule (HPA: HPA005909 ICC Supported; subcellular). |
| Situation | Likely cause | Next action |
|---|---|---|
| No color in an HPA High cell population (HPA: tissue IHC). | A failed staining run, unsuitable control, or weak detection is possible (standard IHC practice). | Check a same-run positive tissue and reagent controls, then review the antibody dilution and chromogen steps used in the run (standard IHC practice). |
| Only adipocytes or cardiomyocytes stain strongly (HPA: Not detected in these cells). | Cell misidentification, cross-reactivity, or endogenous detection activity may account for the discordance (HPA: tissue IHC; standard IHC practice). | Confirm morphology and compare with a no-primary control; if needed, assess endogenous enzyme blocking for the chosen chromogen system (standard IHC practice). |
| The whole section has diffuse brown haze (standard IHC practice). | Nonspecific antibody binding, inadequate washing, or detection background may obscure cell-associated staining (standard IHC practice). | Review blocking and washing, and compare a no-primary control before scoring cytoplasmic signal (standard IHC practice). |
| Signal is restricted to an edge or damaged area (standard IHC practice). | Section handling or local reagent accumulation can produce uneven color (standard IHC practice). | Assess intact interior tissue and repeat the stain if the artifact prevents cell-level scoring (standard IHC practice). |
| Prominent nuclear color appears without cytoplasmic color (HPA: general cytoplasmic expression). | Nuclear TRIM25 is possible, but HPA's nucleoplasmic ICC-IF location is uncertain; detection background is also possible (UniProt Q14258: nucleus; HPA: nucleoplasm uncertain; standard IHC practice). | Record nuclear and cytoplasmic compartments separately, check controls, and avoid calling the nuclear-only result the expected tissue IHC pattern (HPA: general cytoplasmic expression; standard IHC practice). |
| The positive control works, but a study section is negative (standard IHC practice). | The sampled cell population may differ from an HPA High population; HPA reports low tissue specificity and medium staining-to-RNA consistency (HPA: RNA specificity; reliability description). | Identify the cells being scored, document the negative result, and compare it with an HPA cell-level pattern without assuming a target-specific fixation effect (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing TRIM25 staining
M03232 · IHCIHC analysis of TRIM25 using anti-TRIM25 antibody (M03232). TRIM25 was detected in a paraffin-embedded section of human breast cancer tissue
M03232 · IHCIHC analysis of TRIM25 using anti-TRIM25 antibody (M03232). TRIM25 was detected in a paraffin-embedded section of human breast cancer tissue
A03232-1 · IHCIHC analysis of TRIM25 using anti-TRIM25 antibody (A03232-1). TRIM25 was detected in paraffin-embedded section of human mammary cancer tissu
A03232-1 · IHCIHC analysis of TRIM25 using anti-TRIM25 antibody (A03232-1). TRIM25 was detected in paraffin-embedded section of human mammary cancer tissu
A03232-1 · IFIF analysis of TRIM25 using anti-TRIM25 antibody (A03232-1). TRIM25 was detected in immunocytochemical section of U20S cells. Enzyme antigen
A03232-2 · IFImmunocytochemistry of TRIM25 in HeLa cells with TRIM25 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000121060-TRIM25/tissue/cerebral+cortex#imid_1870400
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000121060-TRIM25/tissue/colon#imid_1870326
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000121060-TRIM25/tissue/liver#imid_1870335
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000121060-TRIM25/tissue/kidney#imid_1870328
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005909; green = TRIM25, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005909; green = TRIM25, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TRIM25 IHC & IF
🧪Run breast first: glandular cells should stain (HPA: High in breast glandular cells). Use adipose tissue as the negative tissue: adipocytes should be unstained (HPA: Not detected in adipocytes); on the breast slide, assess any morphologically distinct non-glandular cells as internal background references, without assuming they are TRIM25-negative.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TRIM25 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (uncertain), Cytosol (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and rabbit IgG isotype controls matched to the primary antibody’s clonality, plus a TRIM25 knockout specimen as a biological specificity control (caption: rabbit anti-TRIM25 antibody; standard IHC practice). For DAB detection, block endogenous peroxidase and check the breast section for residual background (caption: peroxidase secondary and DAB).
⚠️Feasibility: The selected M03232 paraffin-section caption does not report a fixative, and the supplied evidence reports no TRIM25-specific fixation window or fixation effect (M03232 caption). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that this retrieval method is required (M03232 caption). Frozen sections or IF cannot be judged easier from the supplied evidence; no breast-specific artefact is reported, so assess background with the controls above (M03232 caption; HPA subcellular: ICC-IF images available).
HPA tissue IHC evidence for TRIM25
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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | Endothelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TRIM25 IHC Tips
Troubleshoot TRIM25 staining in paraffin sections by checking retrieval, compartment, cell identity and controls before interpreting chromogenic signal.
What should I change when TRIM25 staining is weak after retrieval?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet M03232). The documented breast cancer section used this retrieval before overnight primary incubation at 4°C, so compare weak runs with that sequence (datasheet M03232). If staining remains weak, vary heating duration on matched sections while holding antibody concentration, detection and development constant; record tissue preservation alongside signal (standard IHC practice). Include a known positive cell population, such as breast glandular cells, and a primary-omission control to distinguish retrieval failure from background (HPA: High in breast glandular cells; standard IHC practice). Avoid interpreting stronger DAB alone as improved specificity (standard IHC practice).
Can fixation explain absent TRIM25 staining in my paraffin sections?
TRIM25-specific sensitivity to fixation is unknown from the supplied evidence: the selected paraffin-section caption does not state a fixative (datasheet M03232). Record the actual fixative, fixation duration and processing history for each specimen before comparing staining across batches (standard IHC practice). When signal is absent, stain matched sections from a known positive tissue in the same run and compare preservation, retrieval and primary-antibody incubation (HPA: High in breast glandular cells; datasheet M03232; standard IHC practice). Examine nuclei and morphology after counterstaining, because damaged sections make compartment scoring unreliable (standard IHC practice). Do not assign a TRIM25-specific fixation mechanism without direct evidence (datasheet M03232).
Should TRIM25 appear in nuclei, cytoplasm or both by IHC?
Expect predominantly cytoplasmic staining in tissue sections, while allowing a nuclear component (HPA tissue IHC: general cytoplasmic expression; UniProt Q14258: cytoplasm and nucleus). Cytosolic localisation is supported in cell imaging, whereas nucleoplasmic and nuclear-body assignments are uncertain; those imaging observations are context for reviewing IHC, not proof of a nuclear IHC pattern (HPA subcellular). Compare cytoplasmic and nuclear DAB separately against the counterstain and score only intact cells with discernible boundaries (standard IHC practice). If puncta are prominent, consider the reported stress-granule localisation but check whether the pattern recurs in positive tissue and disappears in controls (UniProt Q14258: stress granule; standard IHC practice).
How can I assess an unexpected TRIM25 staining pattern without an epitope map?
The supplied record lists 0 isoforms and a 1–630 chain, but gives no binding epitope for M03232 (UniProt Q14258; datasheet M03232). TRIM25 contains a B30.2/SPRY domain at residues 439–630 and several annotated modified residues; neither fact establishes which region this antibody recognises (UniProt Q14258). Do not attribute compartment differences or lost staining to an isoform or masked modification on this evidence alone (UniProt Q14258; standard IHC interpretation). Compare serial sections with an independently validated antibody whose epitope is documented, alongside omission controls, before treating discordant patterns as biology (standard IHC practice). Record retrieval and processing conditions for that comparison (standard IHC practice).
How should I investigate TRIM25 localisation with multiplex IF?
Use IF/ICC as a separate localisation experiment; the M03232 evidence here documents paraffin-section chromogenic IHC, not an IF dilution or fixation condition (datasheet M03232). Multiplex TRIM25 with a validated marker for the expected cell population, such as breast glandular cells, and include single-stain controls to assess channel bleed-through (HPA: High in breast glandular cells; standard IF practice). Select a fluorophore whose emission is separated from measured tissue autofluorescence, and inspect unstained tissue before assigning dim puncta (standard IF practice). Because TRIM25 has no transmembrane segment and is reported in cytoplasm and nucleus, optimise permeabilisation for intracellular access without assuming an extracellular epitope (UniProt Q14258 topology and localisation; standard IF practice).
What causes widespread brown signal in TRIM25 IHC?
First compare the stained section with a primary-omission control and a known low-expression cell population to locate reagent and tissue background (standard IHC practice; HPA: TRIM25 not detected in adipocytes). Block endogenous peroxidase before HRP detection and assess pigment or precipitate under the counterstain, since both can resemble DAB signal (standard IHC practice). The documented M03232 section used 10% goat serum blocking, a 1:50 primary incubation overnight at 4°C, and peroxidase detection with DAB (datasheet M03232). If background rises with longer development, shorten DAB exposure on matched sections while preserving the same positive control and retrieval conditions (standard IHC practice).
How should I quantify TRIM25 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before reading slides, because tissue IHC is generally cytoplasmic while nuclear localisation is also reported (HPA tissue IHC; UniProt Q14258 localisation). For a defined population, report the percentage of positive cells and staining intensity, or calculate an H-score from intensity categories; keep thresholds fixed across the batch (standard IHC practice). If counting discrete positive cells, report density per mm² of viable tissue and normalise to the eligible cell population or viable area rather than whole-slide area (standard IHC practice). Exclude folds, necrosis and section edges, and review weak staining against a concurrent control (standard IHC practice).
When is TRIM25 staining credible rather than an artefact?
A credible pattern should recur in intact cells of a plausible population and broadly match cytoplasmic TRIM25 expression; nuclear signal remains biologically possible (HPA tissue IHC; UniProt Q14258 localisation). Breast glandular cells provide a documented high-staining reference, whereas adipocytes are reported as not detected in HPA tissue IHC (HPA: High in breast glandular cells; HPA: Not detected in adipocytes). Treat staining confined to edges, necrosis or crushed areas cautiously, and compare it with primary-omission and endogenous-peroxidase controls (standard IHC practice). If signal appears only in an unexpected compartment or cell type, verify cell identity and repeat staining on matched sections before assigning a TRIM25-specific change (standard IHC practice).
Boster reagents
Best TRIM25 / E3 ubiquitin/ISG15 ligase TRIM25 IHC Antibodies
Anti-TRIM25 antibodies have IHC images from human breast and pancreatic cancer sections and mouse stomach sections, plus IF/ICC images from human cells (catalog image captions).
Real IHC data
Anti-TRIM25 Monoclonal Antibody
Cat # M03232
Real IHC data
Anti-TRIM25/EFP Antibody ®
Cat # A03232-1
Real IF data
Anti-TRIM25 Antibody
Cat # A03232-2
M03232 has IHC images from human breast cancer and mouse stomach paraffin sections; A03232-1 has IHC images from human mammary and pancreatic cancer paraffin sections and an IF image from U20S cells (catalog image captions). A03232-2 has IF/ICC images from HeLa cells and lists human and mouse reactivity, with no IHC application listed (catalog applications, reactivity, and image captions).
Which to pick: For human tissue IHC, choose A03232-1; its own captions show paraffin sections stained after EDTA retrieval at pH 8.0 with 1 μg/mL primary antibody (A03232-1 IHC image captions). For IF/ICC, choose A03232-2 based on its HeLa cell images at 20 μg/mL; for IHC across species, consider monoclonal M03232, which lists human, mouse, and rat reactivity and shows human and mouse paraffin sections stained after EDTA retrieval at pH 8.0 with a 1:50 dilution (A03232-2 catalog and IF/ICC captions; M03232 catalog and IHC image captions). Fixative is unreported for both antibodies’ IHC images, and the M03232 captions do not show rat tissue (A03232-1 and M03232 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q14258 (TRI25_HUMAN, E3 ubiquitin/ISG15 ligase TRIM25).
- Human Protein Atlas. TRIM25 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. TRIM25 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and cytosol. In addition localized to the nuclear bodies..
- Human Protein Atlas. TRIM25 antibody validation summary (1 antibodies).
- TRIM25-mediated ferroptosis resistance is closely associated with poor prognosis in hepatocellular carcinoma. Frontiers in immunology 2026 — PMC13079618.
- TRIM25-Mediated Ubiquitination and Degradation of SOX8 Promotes Ligament Fibroblast Osteogenic Differentiation and Regulates OPLL Progression by Inhibiting OSR2 Transcription. JOR spine 2025 — PMC12412890.
- Identification and Validation of TRIM25 as a Glucose Metabolism Regulator in Prostate Cancer. International journal of molecular sciences 2022 — PMC9408812.
- The ubiquitin ligase TRIM25 targets ERG for degradation in prostate cancer. Oncotarget 2016 — PMC5323126.
- PubMed PMID:8248217 — UniProt-cited evidence.
- PubMed PMID:16625196 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TRIM25 (E3 ubiquitin/ISG15 ligase TRIM25). Expected localisation General cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03232). Validated IHC antibody: M03232 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet M03232); 2. Fixation: Image fixative and duration unreported (datasheet M03232); 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 M03232); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03232); 8. Primary antibody: Rabbit monoclonal (clone ADFG-20) anti-TRIM25, 1:50 (datasheet M03232); 9. Primary incubation: Overnight at 4 °C (datasheet M03232). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03232); Caveat: Endogenous peroxidase in marrow may mimic DAB staining (HPA tissue IHC; standard IHC practice); Isoform / epitope: No annotated isoforms or processing to alter the epitope map (UniProt); Regulation: Staining-linked regulation unreported (HPA tissue IHC). Evidence: UniProt Q14258, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and cytosol. In addition…, Datasheet M03232, PMC13079618, PMC9408812. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TRIM25/immunohistochemistry-ihc-guide.