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IDH2 / Isocitrate dehydrogenase [NADP], mitochondrial · IHC design guide
Design Immunohistochemistry for IDH2
Plan IDH2 chromogenic IHC in paraffin sections around its granular cytoplasmic tissue pattern (HPA tissue IHC) and mitochondrial location (UniProt). Use the documented positive and undetected cell populations to assess staining, with attention to endogenous peroxidase in bone marrow (HPA tissue IHC; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
IDH2 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 | Granular cytoplasm (HPA tissue IHC); mitochondrion (UniProt) | |
| Staining pattern | General granular cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00510-3) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Epididymis+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 | Endogenous peroxidase in bone marrow may confound staining (HPA tissue IHC; standard IHC practice) | |
| Regulation | Tissue-dependent expression (HPA tissue IHC) | |
| Isoform / epitope | 2 isoforms; mature chain 40–452; epitope coverage unknown (UniProt) |
Section 1
Recommended IDH2 IHC & IF Protocols
The catalog antibody's IHC-P protocol is paired with a published protocol for IDH2 R172S staining in breast tumors (PMC7286791).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M00510-3) |
| Fixation | Image fixative and duration unreported (datasheet M00510-3); 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 M00510-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00510-3) |
| Primary antibody | Mouse monoclonal (clone 2H4) anti-IDH2, 2μg/ml (datasheet M00510-3) |
| Primary incubation | Overnight at 4 °C (datasheet M00510-3) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet M00510-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | IDH2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet M00510-3). The published R172S assay used high-pH retrieval for 30 minutes (PMC7286791).
Section 2
What Is the Expected IDH2 Staining Pattern?
IDH2 localizes to mitochondria and has no transmembrane segment (UniProt P48735). In paraffin sections, expect granular cytoplasmic staining in positive cells, including colon glandular cells and bronchial respiratory epithelial cells (HPA: High; general cytoplasmic, granular pattern). HPA rates the tissue IHC evidence Enhanced, while reporting medium consistency between antibody staining and RNA expression data (HPA: reliability).
| Granular cytoplasmic staining in colon glandular cells or bronchial respiratory epithelial cells. | This fits the reported pattern in these High staining cell populations (HPA: tissue IHC). Granularity is consistent with mitochondrial localization (UniProt P48735; HPA: subcellular), although chromogenic IHC alone does not prove that each visible granule is a mitochondrion (general IHC practice). |
| Predominantly nuclear or sharply cell-surface staining, with little granular cytoplasmic signal. | That compartment conflicts with the reported mitochondrial localization and general granular cytoplasmic IHC profile (UniProt P48735; HPA: tissue IHC and subcellular). Review the signal as a possible staining artefact; compartment alone cannot identify its cause (general IHC practice). |
| Strong staining in liver cholangiocytes, while the expected positive cells are weak. | HPA reports IDH2 as Not detected in cholangiocytes, specifically; it does not classify every liver cell as negative (HPA: tissue IHC). Consider cross-reactivity or endogenous chromogenic activity, then compare cell identity and controls (general IHC practice). |
| Uniform color across cells, extracellular areas, or most of the section. | A widespread haze obscures the reported granular cytoplasmic pattern (HPA: tissue IHC). It can arise from nonspecific staining or the detection system; judge it against a control processed with the same detection reagents (general IHC practice). |
| No staining in colon glandular cells or bronchial respiratory epithelial cells. | Those cells are reported High, so an absent signal calls for a technical review (HPA: tissue IHC). Check tissue preservation, retrieval, antibody application, and detection with appropriate controls before interpreting the result as biological absence (general IHC practice). |
💡Expected IDH2 appearanceCall a positive result when identified High staining cells show clear granular cytoplasmic color (HPA: tissue IHC); dominant nuclear, cell-surface, or diffuse section-wide color is a possible false positive pattern given IDH2's mitochondrial location (UniProt P48735; HPA: subcellular; general IHC practice).
| Cell identity and comparator | HPA reports High staining in colon glandular cells and Not detected staining in liver cholangiocytes (HPA: tissue IHC). Score the named cell populations; neither observation defines an entire organ as uniformly positive or negative (general IHC practice). |
| Compartment and topology | IDH2 is mitochondrial and has no annotated transmembrane segment (UniProt P48735). The matching tissue IHC description is general granular cytoplasmic expression (HPA: tissue IHC); a continuous plasma-membrane outline is therefore unexpected. |
| Antibody evidence | HPA lists HPA007831 as Enhanced for IHC and Supported for ICC, and describes medium consistency between staining and RNA expression (HPA: antibody validation; tissue IHC reliability). These ratings support use of the reported pattern while leaving individual slide interpretation dependent on controls. |
| Processing and isoforms | UniProt lists two isoforms and a mature chain spanning residues 40–452 (UniProt P48735). The supplied record gives no antibody epitope, so it cannot establish whether either isoform or processing changes this antibody's staining; avoid assigning a compartment change to either. |
| IF/ICC Q&A: what localization should an image show? | The supported IF/ICC location is mitochondria; HPA lists images for A-431, U-251MG, and U2OS and rates HPA007831 ICC Supported (HPA: subcellular; antibody validation). This is a localization answer, not an IF/ICC protocol or a claim about paraffin-section signal intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cells remain unstained. | The run may have lost detectable signal; the supplied sources do not establish IDH2-specific fixation sensitivity (HPA: tissue IHC; UniProt P48735). | Confirm the named positive cell population, then review the run's retrieval, primary antibody, and detection controls using general IHC practice. Treat an isolated negative slide cautiously when HPA reports those cells High (HPA: tissue IHC). |
| Color is predominantly nuclear or outlines cell surfaces. | The distribution disagrees with mitochondrial localization and the reported granular cytoplasmic profile (UniProt P48735; HPA: tissue IHC). | Verify morphology and repeat with appropriate negative and positive controls. Review nonspecific binding or detection artefact before scoring it as IDH2 (general IHC practice). |
| A Not detected comparator stains strongly. | For example, cholangiocytes are reported Not detected; unexpected color could reflect cell misidentification, cross-reactivity, or detection background (HPA: tissue IHC; general IHC practice). | Confirm that the stained cells are cholangiocytes and compare them with a High staining population and detection controls. Do not treat the whole liver as the negative comparator (HPA: tissue IHC). |
| Granular signal is buried in diffuse brown haze. | Background from nonspecific binding or endogenous detection activity can obscure cell-level interpretation (general IHC practice). | Inspect a matched negative detection control and the blocking and wash steps. Assess endogenous peroxidase control if using peroxidase detection; then judge whether the granular cytoplasmic pattern is distinguishable (general IHC practice; HPA: tissue IHC pattern). |
| Signal varies between cells or sections. | HPA reports specific cell populations at different levels, including High colon glandular cells and Not detected liver cholangiocytes (HPA: tissue IHC). Variation alone does not show an assay failure. | Record tissue, cell identity, compartment, and intensity separately. Compare like cell populations and the same run's controls before attributing variation to staining conditions (general IHC practice). |
| The researcher wants to transfer this chromogenic result to IF/ICC. | HPA supports mitochondrial ICC localization, while this section's tissue intensity categories come from IHC (HPA: subcellular; tissue IHC). | Use the IF/ICC guide for its own controls and imaging decisions. Expect a mitochondrial localization pattern in supported ICC evidence, but do not convert IHC intensity categories into IF intensity predictions (HPA: subcellular; general IF practice). |
Experiment evidence
IHC & IF images showing IDH2 staining
M00510-3 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510-3). IDH2 was detected in paraffin-embedded section of human lung cancer tissue. Heat m
M00510-3 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510-3). IDH2 was detected in paraffin-embedded section of human melanoma tissue. Heat medi
M00510-4 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510-4). IDH2 was detected in paraffin-embedded section of human testicular cancer tissue.
M00510-4 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510-4). IDH2 was detected in paraffin-embedded section of human rectal cancer tissue. Heat
M00510-4 · IFIF analysis of IDH2 using anti-IDH2 antibody (M00510-4). IDH2 was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
PB9602 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (PB9602). IDH2 was detected in a paraffin-embedded section of human colon cancer tissue. Heat
PB9602 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (PB9602). IDH2 was detected in a paraffin-embedded section of human ovarican cancer tissue. He
PB9602 · IFIF analysis of IDH2 using anti-IDH2 antibody (PB9602). IDH2 was detected in an immunocytochemical section of A549 cells. Enzyme antigen retr
M00510 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510). IDH2 was detected in a paraffin-embedded section of human colon cancer tissue. Heat
M00510 · IHCIHC analysis of IDH2 using anti-IDH2 antibody (M00510). IDH2 was detected in a paraffin-embedded section of human colon cancer tissue. Heat
A00510 · IHCImmunohistochemistry of IDH2 in mouse heart tissue with IDH2 antibody at 5 μg/mL.
A00510 · IFImmunofluorescence of IDH2 in mouse heart tissue with IDH2 antibody at 20 μg/mL.
M00510-2 · IFIF analysis of IDH2 using anti-IDH2 antibody (M00510-2). IDH2 was detected in immunocytochemical section of A431 cells. Enzyme antigen retri
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182054-IDH2/tissue/cerebral+cortex#imid_2357292
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182054-IDH2/tissue/colon#imid_2357218
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182054-IDH2/tissue/liver#imid_2357227
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000182054-IDH2/tissue/kidney#imid_2357220
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007831; green = IDH2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA007831; green = IDH2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for IDH2 IHC & IF
🧪Run appendix first: its glandular cells should stain strongly (HPA: High in appendix glandular cells). Use epididymis glandular cells as a negative tissue comparator (HPA: Not detected in epididymis glandular cells); on the appendix slide, treat cells with absent staining as an internal background reference only after identifying them, since the supplied HPA row specifies glandular cells but does not classify adjacent cells (HPA: High in appendix glandular cells).
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Epididymis (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show IDH2 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host- and isotype-matched control antibody sections; the selected protocol uses a mouse primary (M00510-3 tissue-IHC caption). Confirm specificity with matched IDH2-knockout material where available, or a validated immunizing-peptide block; quench endogenous peroxidase and check endogenous biotin background because the caption uses a biotin-based detection system with DAB (M00510-3 tissue-IHC caption).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the fixative in the selected paraffin-section caption is unreported (M00510-3 tissue-IHC caption). That caption reports heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is essential; the supplied evidence does not establish that frozen sections or IF are easier, although mitochondrial ICC-IF localization is supported (M00510-3 tissue-IHC caption; HPA: Mitochondria, supported). No appendix-specific artefact is documented in the supplied evidence; inspect biotin-associated background when using the caption’s detection system (M00510-3 tissue-IHC caption).
HPA tissue IHC evidence for IDH2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | Ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced IDH2 IHC Tips
These questions guide chromogenic IHC of paraffin sections stained for IDH2, with one entry on adapting the readout to IF.
What retrieval should I try first if IDH2 staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M00510-3). The selected tissue image used that condition before overnight incubation at 4°C with 2 µg/mL antibody (datasheet M00510-3). If staining remains weak, vary heating time in small steps while keeping the buffer fixed, and compare sections processed in the same run (standard IHC practice). Check that stronger signal retains a granular cytoplasmic pattern and does not merely increase diffuse staining or damaged tissue edges (HPA tissue IHC profile; standard IHC practice).
How should I investigate variable staining across differently fixed samples?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (datasheet M00510-3). Record each specimen’s fixative, fixation duration, processing history, and section age before comparing staining, because these variables can affect antigen accessibility in paraffin IHC (standard IHC practice). Run differently processed specimens together with the same EDTA pH 8.0 retrieval and antibody conditions to identify workflow-related variation (datasheet M00510-3; standard IHC practice). Do not attribute a loss of IDH2 signal to a particular fixative without a controlled comparison using the same tissue and detection workflow (standard IHC practice).
Which staining pattern supports a specific IDH2 result?
Look for granular cytoplasmic staining consistent with mitochondrial localisation when evaluating chromogenic sections (HPA tissue IHC profile; HPA subcellular; UniProt P48735). Compare the pattern within cells and across compartments: predominantly nuclear or extracellular DAB should prompt review of counterstain, tissue pigment, and antibody background (UniProt P48735 localisation; standard IHC practice). Bronchial respiratory epithelial cells and colonic glandular cells provide documented high-staining examples, while liver cholangiocytes are listed as not detected (HPA tissue IHC). Use these references to check pattern plausibility, while interpreting each specimen’s signal against its own tissue morphology and staining controls (standard IHC practice).
Can this stain distinguish IDH2 isoforms or processing states?
Do not assign isoform identity from this chromogenic stain alone: the record lists 2 isoforms, but the supplied antibody caption does not map its epitope (UniProt P48735; datasheet M00510-3). The annotated mature chain spans residues 40–452, and acetylated residues include 45 and 48 (UniProt P48735). Without an epitope map, neither precursor recognition nor sensitivity to those modifications can be inferred from the tissue image (UniProt P48735; datasheet M00510-3). For an isoform-specific or modification-specific claim, obtain the antibody’s epitope information and validate it with appropriate reference material before interpreting section scores (standard IHC practice).
How would I assess IDH2 in a multiplex IF experiment?
Treat transfer of this antibody from chromogenic paraffin IHC to IF as a separate validation task; the selected caption documents tissue IHC conditions only (datasheet M00510-3). Pair IDH2 with a validated respiratory epithelial cell marker when examining bronchial epithelium, where high staining is reported, and include single-stain controls for channel separation (HPA tissue IHC; standard IF practice). Choose a fluorophore away from the specimen’s strongest autofluorescence and check unstained tissue before assigning faint puncta to IDH2 (standard IF practice). Because IDH2 is mitochondrial and has no annotated transmembrane segment, optimise permeabilisation for antibody access to an internal mitochondrial epitope while checking mitochondrial morphology (UniProt P48735; standard IF practice).
How can I separate weak IDH2 staining from detection background?
The selected workflow uses a biotinylated secondary, streptavidin–biotin detection, and DAB, so inspect background at each detection step (datasheet M00510-3; standard IHC practice). Include a no-primary control to reveal signal from the secondary or detection reagents, and assess endogenous biotin when using this detection chemistry (standard IHC practice). Apply a peroxidase block before DAB development and compare stained sections with the no-primary control under identical development conditions (standard IHC practice). If background persists, optimise antibody concentration around the documented 2 µg/mL condition while preserving the expected granular cytoplasmic signal (datasheet M00510-3; HPA tissue IHC profile; standard IHC practice).
How should I score IDH2 across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score only viable cells with interpretable granular cytoplasmic staining, using matched staining and imaging conditions across sections (HPA tissue IHC profile; standard IHC practice). An H-score combines the percentage of cells at each intensity grade 0–3 into a 0–300 score; record the percentage positive separately to make distribution changes visible (standard IHC practice). Normalise counts or stained area to the number of eligible cells or measured viable tissue area, and report results separately by cell type (standard IHC practice). Document staining controls and section quality because HPA reports only medium consistency between antibody staining and RNA expression data (HPA tissue IHC).
Which apparent positives should I question before reporting IDH2 expression?
A credible positive should fit IDH2’s mitochondrial localisation and the granular cytoplasmic tissue pattern, with signal inside morphologically intact cells (UniProt P48735; HPA subcellular; HPA tissue IHC profile). Reconsider predominantly nuclear or extracellular signal, abrupt staining confined to section edges, and dense DAB over necrotic regions; inspect adjacent morphology and the no-primary control (standard IHC practice). Check cell identity before scoring: HPA lists high staining in bronchial respiratory epithelial cells but no detection in liver cholangiocytes (HPA tissue IHC). Finally, exclude endogenous peroxidase or biotin-related detection signal using workflow controls before calling a weak chromogenic deposit IDH2 (standard IHC practice).
Boster reagents
Best IDH2 / Isocitrate dehydrogenase [NADP], mitochondrial IHC Antibodies
Catalog images document IDH2 IHC in human cancer paraffin sections and mouse heart tissue, plus IF in A431 cells (catalog image captions: M00510-3, M00510-4, PB9602, M00510, A00510, M00510-2).
Real IHC data
Anti-IDH2 Antibody ® (monoclonal, 2H4)
Cat # M00510-3
Real IHC data
Anti-IDH2 Antibody ® (monoclonal, 6B13)
Cat # M00510-4
Real IHC data
Anti-IDH2 Antibody ®
Cat # PB9602
Real IHC data
Anti-IDH2 Monoclonal Antibody
Cat # M00510
Real IHC data
Anti-IDH2 Antibody
Cat # A00510
Real IF data
Anti-IDH2 Antibody ® (monoclonal, 2D4)
Cat # M00510-2
IHC figures show M00510-3 in human lung cancer, M00510-4 in human testicular cancer, PB9602 and M00510 in human colon cancer, and A00510 in mouse heart (catalog image captions). M00510-2 has an IF figure in A431 cells (catalog image caption).
Which to pick: For paraffin-section IHC, consider M00510-3 or M00510-4: both are mouse monoclonals whose own captions show EDTA retrieval at pH 8.0 and primary antibody at 2 μg/mL in human lung or testicular cancer sections, respectively; the fixative is unreported (catalog clone fields and IHC image captions). For IF/ICC, M00510-2 has an A431 cell IF figure, while M00510-4 lists both IF/ICC and IHC applications (catalog image captions and applications). For IHC across species, PB9602 has images from human colon cancer and mouse and rat heart, with Human, Mouse and Rat reactivity listed; its own IHC caption reports EDTA retrieval at pH 8.0 and 2 μg/mL primary antibody, but no fixative (catalog IHC image captions and reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P48735 (IDHP_HUMAN, Isocitrate dehydrogenase [NADP], mitochondrial).
- Human Protein Atlas. IDH2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. IDH2 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. IDH2 antibody validation summary (1 antibodies).
- Immunohistochemical analysis of IDH2 R172 hotspot mutations in breast papillary neoplasms: applications in the diagnosis of tall cell carcinoma with reverse polarity. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2020 — PMC7286791.
- The role of a monoclonal antibody 11C8B1 as a diagnostic marker of IDH2-mutated sinonasal undifferentiated carcinoma. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 2019 — PMC7429919.
- Isocitrate dehydrogenase 2 mutation is a frequent event in osteosarcoma detected by a multi-specific monoclonal antibody MsMab-1. Cancer medicine 2013 — PMC3892385.
- Molecular epidemiology of IDH2 hotspot mutations in cancer and immunohistochemical detection of R172K, R172G, and R172M variants. Human pathology 2020 — PMC7721993.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16572171 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for IDH2 (Isocitrate dehydrogenase [NADP], mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrion (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00510-3). Validated IHC antibody: M00510-3 at 2μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Epididymis. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet M00510-3); 2. Fixation: Image fixative and duration unreported (datasheet M00510-3); 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 M00510-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00510-3); 8. Primary antibody: Mouse monoclonal (clone 2H4) anti-IDH2, 2μg/ml (datasheet M00510-3); 9. Primary incubation: Overnight at 4 °C (datasheet M00510-3). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Endogenous peroxidase in bone marrow may confound staining (HPA tissue IHC; standard IHC practice); Isoform / epitope: 2 isoforms; mature chain 40–452 ; epitope coverage unknown (UniProt); Regulation: Tissue-dependent expression (HPA tissue IHC). Evidence: UniProt P48735, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet M00510-3, PMC7286791. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/IDH2/immunohistochemistry-ihc-guide.