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TRIM2 / Tripartite motif-containing protein 2 · IHC design guide
Design Immunohistochemistry for TRIM2
Plan paraffin-section TRIM2 IHC using cerebellar GLUC cells as a high-staining reference and adipocytes as a negative cellular control (HPA tissue IHC). Start the IHC-validated antibody A08957 at 2–5 μg/ml (datasheet A08957), then assess cytoplasmic staining with the reported off-target binding in mind (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TRIM2 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 | Mainly cytoplasmic staining in the CNS (HPA tissue IHC) | |
| Staining pattern | CNS cytoplasm; cerebellar GLUC cells also stain at the membrane (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A08957) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Presumed off-target binding may mimic TRIM2 staining (HPA tissue IHC) | |
| Regulation | No stimulus-dependent regulation annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage for each (UniProt) |
Section 1
Recommended TRIM2 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet: A08957) with a published protocol for human breast tumour sections (PMC11171213).
| Sample | Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A08957) |
| Fixation | Image fixative and duration unreported (datasheet A08957); 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 A08957); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A08957) |
| Primary antibody | Rabbit anti-TRIM2, 2-5 μg/ml (datasheet A08957) |
| Primary incubation | Overnight at 4 °C (datasheet A08957) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A08957) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TRIM2-positive staining in gLUC cells - cytoplasm/membrane of cerebellum (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in the CNS. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: A08957); citrate at pH 6.0 is a published alternative (PMC11171213).
Section 2
What Is the Expected TRIM2 Staining Pattern?
TRIM2 is annotated as cytoplasmic and has no transmembrane segment (UniProt Q9C040). In paraffin-section IHC, expect staining mainly in CNS cells: high staining in cerebellar GLUC cells and medium staining in cerebral cortex and hippocampal glial cells (HPA tissue IHC). HPA rates its tissue evidence Supported, with medium agreement between staining and RNA data; its antibody findings need cautious interpretation (HPA tissue IHC).
| Cerebellar GLUC cells stain strongly in the cytoplasm, with possible membrane-associated staining. | This matches the highest supplied tissue observation: GLUC cells show high cytoplasmic/membrane staining (HPA tissue IHC). The cytoplasmic component agrees with TRIM2's annotated location (UniProt Q9C040). Assess the stained cells and compartment together; intensity alone cannot establish antibody specificity (standard IHC practice). |
| Glial cells in cerebral cortex or hippocampus show moderate staining. | Medium glial-cell staining is reported in both regions (HPA tissue IHC). A weaker result than the high cerebellar GLUC-cell observation can therefore fit the supplied pattern (HPA tissue IHC). Compare like cell populations within the section when scoring, since tissue-wide color can obscure which cells carry the signal (standard IHC practice). |
| Signal is predominantly nuclear, or forms a convincing isolated nuclear pattern. | Treat this as discordant with the annotated cytoplasmic location (UniProt Q9C040). Review morphology and controls before calling it TRIM2; misplaced signal can reflect nonspecific staining or detection artefact (standard IHC practice). The uncertain centrosome call from ICC-IF does not establish an expected nuclear IHC pattern (HPA subcellular ICC-IF). |
| Strong staining appears in cells listed as not detected, such as adipocytes or bone-marrow hematopoietic cells. | Those cell populations are reported as not detected in the supplied tissue observations (HPA tissue IHC). Check for cross-reactivity or endogenous detection activity before interpreting a strong result as TRIM2 (standard IHC practice). HPA also cautions that its tissue antibodies target proteins from more than one gene and that presumed off-target binding was disregarded (HPA tissue IHC). |
| Color is diffuse across tissue structures, or a known-positive region has no discernible signal. | Diffuse color without cell-specific contrast is background rather than a persuasive localisation pattern (standard IHC practice). No signal in cerebellar GLUC cells conflicts with their reported high staining (HPA tissue IHC), but does not by itself prove TRIM2 absence. Compare a positive-control section and detection controls before assigning a biological negative (standard IHC practice). |
💡Expected TRIM2 appearanceCall a positive when cerebellar GLUC cells show high cytoplasmic/membrane staining, with medium glial staining plausible in cortex or hippocampus (HPA tissue IHC); widespread nuclear or cell-independent color is suspect (UniProt Q9C040; standard IHC practice).
| Compartment and topology | TRIM2 is annotated in the cytoplasm and has no transmembrane segment (UniProt Q9C040). Interpret the HPA cerebellar cytoplasmic/membrane observation as a cell staining pattern, without inferring that TRIM2 spans the membrane (HPA tissue IHC; UniProt Q9C040). |
| Tissue and cell selection | Cerebellar GLUC cells provide the supplied high-staining comparison; cortex and hippocampus provide medium glial-cell observations (HPA tissue IHC). Adipocytes and bone-marrow hematopoietic cells are listed as not detected, so use their stated cell identities when evaluating contrast (HPA tissue IHC). |
| Strength of antibody evidence | Two listed antibodies have Supported IHC status, HPA035853 and HPA035854 (HPA antibodies). HPA describes medium consistency with RNA data and cautions about proteins from more than one gene and disregarded presumed off-target binding (HPA tissue IHC). Treat morphology and controls as part of interpretation (standard IHC practice). |
| Isoforms and processing | Two isoforms are listed, with no signal peptide, propeptide, or cleavage into separate annotated chains (UniProt Q9C040). No epitope map is supplied, so these facts cannot predict whether the IHC-validated antibody distinguishes isoforms or whether a particular compartmental signal belongs to one isoform. |
| IF/ICC Q: Should a centrosome spot be expected in tissue IHC? | A: HPA lists centrosome localisation as uncertain in ICC-IF and cautions that the antibodies target proteins from multiple genes (HPA subcellular ICC-IF). Use the cytoplasmic tissue IHC observations for this paraffin-section readout; a centrosome spot alone is insufficient to identify TRIM2 (HPA tissue IHC; HPA subcellular ICC-IF). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in cerebellar GLUC cells. | The result conflicts with the reported high staining in those cells (HPA tissue IHC); a failed staining run or missed cell population is possible (standard IHC practice). | Confirm the cells on the counterstained section, then check a known-positive section, antibody application, and detection controls (standard IHC practice). |
| Only nuclei stain strongly. | A predominantly nuclear result differs from the cytoplasmic annotation (UniProt Q9C040); nonspecific signal or detection artefact is possible (standard IHC practice). | Check a no-primary control and review cell boundaries before scoring; repeat with the IHC-validated antibody if controls implicate the staining run (standard IHC practice). |
| Strong signal appears in an HPA not-detected cell population. | Cross-reactivity or endogenous detection activity may explain unexpected staining (standard IHC practice); HPA notes a multi-gene antibody-target caution (HPA tissue IHC). | Verify the precise cell type, inspect a no-primary control, and assess the relevant detection-system control before calling the cells positive (standard IHC practice). |
| Diffuse brown color obscures cell boundaries. | Non-specific binding or detection background can reduce cell-specific contrast (standard IHC practice). | Compare with a no-primary control; review blocking, antibody concentration, washes, and chromogen development using the staining workflow (standard IHC practice). |
| Cortex or hippocampus looks weaker than cerebellum. | This may match medium glial staining versus high cerebellar GLUC-cell staining (HPA tissue IHC). | Score the named cell populations and their compartments separately; retain the region-specific intensity distinction in the result (HPA tissue IHC; standard IHC practice). |
| A punctate IF/ICC result disagrees with tissue IHC. | The centrosome assignment is uncertain and carries a multi-gene antibody caution (HPA subcellular ICC-IF). | Interpret the paraffin-section result against tissue IHC cell and compartment observations; seek independent specificity evidence before assigning the puncta to TRIM2 (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing TRIM2 staining
A08957 · IHCIHC analysis of TRIM2 using anti-TRIM2 antibody (A08957). TRIM2 was detected in a paraffin-embedded section of human glioblastoma tissue. He
A08957 · IHCIHC analysis of TRIM2 using anti-TRIM2 antibody (A08957). TRIM2 was detected in a paraffin-embedded section of human glioblastoma tissue. He
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000109654-TRIM2/tissue/cerebral+cortex#imid_9089578
HPA · CaudateHuman Protein Atlas IHC, Caudate. Source: https://www.proteinatlas.org/ENSG00000109654-TRIM2/tissue/basal+ganglia#imid_9089564
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000109654-TRIM2/tissue/colon#imid_9089488
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000109654-TRIM2/tissue/liver#imid_9089497
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035854; green = TRIM2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA035854; green = TRIM2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TRIM2 IHC & IF
🧪Run cerebellum first and require cytoplasmic/membrane staining in GLUC cells (HPA: cerebellum GLUC cells, High). Run adipose tissue as a negative, focusing on adipocytes (HPA: adipocytes, Not detected); on the cerebellum slide, treat neighboring cells without the GLUC-cell staining pattern as candidate internal negatives, since their negative status is unreported (HPA: cerebellum GLUC cells, High).
Positive control tissue: Cerebellum (GLUC cells - cytoplasm/membrane, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TRIM2 in A-431, U-251MG, U2OS, with annotated localisation: Centrosome (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched nonimmune rabbit IgG isotype control (caption: rabbit anti-TRIM2 antibody); and TRIM2 knockout tissue or cells as a biological negative. Quench endogenous peroxidase before HRP/DAB detection and check the cerebellum control slide for background color (caption: HRP/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A08957 paraffin-section caption does not state a fixative (caption: fixative not stated). The demonstrated paraffin-section workflow uses heat retrieval in EDTA at pH 8.0 (caption: EDTA retrieval); no matched evidence establishes whether frozen sections or IF are easier. Assess background color in cerebellum alongside the no-primary control before scoring DAB staining (caption: HRP/DAB detection).
HPA tissue IHC evidence for TRIM2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. Presumed off target binding observed and disregarded. Pending external verification.). 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 |
|---|---|---|---|---|
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Medium | Protein (IHC) | HPA → |
| Fallopian tube | Non-ciliated cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TRIM2 IHC Tips
Troubleshoot TRIM2 staining in paraffin sections using the catalog antibody’s IHC example and cautious interpretation of tissue and subcellular evidence.
What retrieval should I try when TRIM2 staining is weak in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A08957). The selected paraffin-section example used that condition before applying 2 μg/ml catalog antibody overnight at 4°C (datasheet A08957). If signal remains weak, check that sections stayed covered during heating and that the positive control stained before changing retrieval conditions (standard IHC practice). A different buffer or pH is a troubleshooting fallback that needs its own control comparison, rather than an established TRIM2 condition (datasheet A08957; standard IHC practice). Score the resulting cytoplasmic signal against a no-primary control (UniProt Q9C040; standard IHC practice).
Could fixation explain weak or uneven TRIM2 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A08957). Record the fixative and processing history for each specimen, then compare sections processed alike when investigating weak or uneven signal (standard IHC practice). Keep retrieval, antibody concentration and detection conditions constant during that comparison; the documented example used EDTA at pH 8.0 and 2 μg/ml antibody (datasheet A08957; standard IHC practice). Uneven staining near section edges calls for checking section handling and reagent coverage, but it does not establish a TRIM2-specific fixation effect (standard IHC practice).
Which compartment should count as TRIM2-positive in chromogenic IHC?
Prioritize staining in the cell body cytoplasm when scoring TRIM2, because its annotated location is cytoplasmic and it has no transmembrane segment (UniProt Q9C040). Tissue IHC describes mainly CNS cytoplasmic expression, including high staining in cerebellar GLUC-cell cytoplasm or membrane and medium staining in cerebral-cortex glial cells (HPA tissue IHC). A centrosome assignment from ICC/IF is uncertain and carries a warning about antibodies targeting proteins from multiple genes (HPA subcellular). Therefore, do not treat isolated puncta or nuclear staining as sufficient evidence of specific TRIM2 in a paraffin section; compare morphology and controls (HPA subcellular; standard IHC practice).
Can this stain distinguish TRIM2 isoforms or modified epitopes?
The record lists 2 TRIM2 isoforms, but the selected tissue-IHC caption supplies no epitope mapping or isoform-specific staining evidence (UniProt Q9C040; datasheet A08957). Interpret catalog-antibody staining as a TRIM2-directed signal without assigning it to isoform 1 or 2 (UniProt Q9C040; datasheet A08957). Several modified residues are annotated, including phosphoserines at positions 10, 375, 424 and 428, yet the caption does not establish whether modification changes epitope recognition (UniProt Q9C040; datasheet A08957). If an isoform or modification claim matters, obtain epitope information and validate the claim with an appropriate independent control (standard IHC practice).
How should I check TRIM2 localisation with multiplex IF?
Use IF as a separate validation experiment and multiplex TRIM2 with a validated marker for the cell population being examined, such as the glial cells reported in cerebral cortex (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and assess each channel against tissue autofluorescence and single-stain controls (standard IF practice). Because TRIM2 is annotated in the cytoplasm and has no transmembrane segment, include a permeabilisation condition that permits antibody access to intracellular epitopes (UniProt Q9C040; standard IF practice). Treat a centrosome-like signal cautiously: the reported ICC/IF location is uncertain and its antibody evidence may include proteins from multiple genes (HPA subcellular).
How can I reduce diffuse brown staining without losing TRIM2 signal?
Run a no-primary control to assess detection background, and inspect a known positive area alongside it before weakening the TRIM2 signal (standard IHC practice; HPA tissue IHC). The documented paraffin-section example blocked with 10% goat serum, used 2 μg/ml primary antibody overnight at 4°C, and developed with peroxidase detection and DAB (datasheet A08957). For diffuse brown signal, check serum blocking, reagent coverage and wash consistency, and include a peroxidase block as a general chromogenic-IHC step (standard IHC practice). Interpret persistent staining cautiously because the tissue-IHC reliability note reports presumed off-target binding and medium RNA-to-staining consistency (HPA tissue IHC).
How should I quantify TRIM2 staining across differently composed sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, using cytoplasmic staining as the primary TRIM2 readout (UniProt Q9C040; standard IHC practice). Within matched regions, report an H-score or the percentage of positive cells, with the denominator set to the eligible cells counted in that same population (standard IHC practice). If counting positive cells per mm², also report the measured tissue area and cell composition so that a shift in density is not mistaken for a change in staining per cell (standard IHC practice). Keep retrieval, detection and scoring thresholds consistent across sections, and interpret comparisons in light of the HPA reliability caution (datasheet A08957; HPA tissue IHC; standard IHC practice).
What distinguishes credible TRIM2 staining from a chromogenic artefact?
Look for cytoplasmic staining in morphologically intact cells, consistent with TRIM2’s annotated location and the mainly CNS cytoplasmic tissue-IHC profile (UniProt Q9C040; HPA tissue IHC). High cerebellar GLUC-cell staining and medium cerebral-cortex glial staining provide contextual comparators, but HPA reports medium RNA-to-staining consistency and presumed off-target binding (HPA tissue IHC). Treat predominantly nuclear signal, staining confined to tissue edges or necrotic areas, and signal persisting without primary antibody as reasons to investigate specificity or processing (UniProt Q9C040; standard IHC practice). Check endogenous peroxidase with the appropriate detection control before attributing brown DAB signal to TRIM2 (standard IHC practice).
Boster reagents
Best TRIM2 / Tripartite motif-containing protein 2 IHC Antibodies
The IHC-validated anti-TRIM2 antibody has paraffin-section images from human glioblastoma and rat brain (A08957 IHC captions); no IF/ICC data are supplied (catalog application list; IF image list).
Real IHC data
Anti-TRIM2 Antibody ®
Cat # A08957
A08957 is the sole listed SKU and has IHC images from paraffin-embedded human glioblastoma and rat brain sections (A08957 IHC captions). Its listed applications include IHC, WB and ELISA, with human, mouse and rat reactivity; IF/ICC is not listed (A08957 catalog).
Which to pick: Choose A08957 for paraffin-section IHC using its documented 2 μg/ml image condition as a starting point; the catalog IHC range is 2–5 μg/ml (A08957 IHC captions; datasheet). No listed SKU has IF/ICC validation, and A08957’s clonality is unspecified (catalog application list; IF image list; clone field). For cross-species work, A08957 lists human, mouse and rat reactivity, while its IHC images document human and rat sections only; the fixative is unreported (A08957 catalog; IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9C040 (TRIM2_HUMAN, Tripartite motif-containing protein 2).
- Human Protein Atlas. TRIM2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. TRIM2 subcellular location (ICC-IF): Localized to the centrosome. Caution: Based on antibodies targeting proteins from multiple genes..
- Human Protein Atlas. TRIM2 antibody validation summary (2 antibodies).
- Interplay of TRIM2 E3 Ubiquitin Ligase and ALIX/ESCRT Complex: Control of Developmental Plasticity During Early Neurogenesis. Cells 2020 — PMC7409263.
- Tripartite Motif-Containing 2, a Glutamine Metabolism-Associated Protein, Predicts Poor Patient Outcome in Triple-Negative Breast Cancer Treated with Chemotherapy. Cancers 2024 — PMC11171213.
- TRIM2 suppresses prostate cancer progression by promoting ubiquitin-mediated degradation of DTNBP1. Frontiers in molecular biosciences 2026 — PMC13616618.
- TRIM2 downregulation in clear cell renal cell carcinoma affects cell proliferation, migration, and invasion and predicts poor patients' survival. Cancer management and research 2018 — PMC6255054.
- PubMed PMID:11331580 — UniProt-cited evidence.
- PubMed PMID:9628581 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TRIM2 (Tripartite motif-containing protein 2). Expected localisation Mainly cytoplasmic staining in the CNS (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08957). Validated IHC antibody: A08957 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (GLUC cells - cytoplasm/membrane); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human glioblastoma tissue; fixative not specified (datasheet A08957); 2. Fixation: Image fixative and duration unreported (datasheet A08957); 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 A08957); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08957); 8. Primary antibody: Rabbit anti-TRIM2, 2-5 μg/ml (datasheet A08957); 9. Primary incubation: Overnight at 4 °C (datasheet A08957). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Presumed off-target binding may mimic TRIM2 staining (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage for each (UniProt); Regulation: No stimulus-dependent regulation annotated (UniProt). Evidence: UniProt Q9C040, HPA tissue IHC · Supported, HPA ICC-IF · centrosome. Caution: Based on antibodies targeting proteins from…, Datasheet A08957, PMC11171213. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TRIM2/immunohistochemistry-ihc-guide.