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CHCHD10 / Coiled-coil-helix-coiled-coil-helix domain-containing protein 10, mitochondrial · IHC design guide
Design Immunohistochemistry for CHCHD10
Plan CHCHD10 IHC in paraffin sections using the granular cytoplasmic tissue profile and high staining in gut endocrine cells and kidney collecting ducts (HPA tissue IHC). The catalog antibody A06312 lists 2–5 μg/mL for human IHC-P (datasheet A06312).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CHCHD10 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 cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Granular cytoplasm; high in gut endocrine cells and kidney collecting ducts (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06312) | |
| Positive control | Appendix+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); target-specific effects are unknown. | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Higher expression in heart and liver (UniProt) | |
| Isoform / epitope | No isoforms listed; processed chain spans residues 17–142 (UniProt) |
Section 1
Recommended CHCHD10 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A06312). The published chromogenic IHC protocol below uses Tris-EDTA pH 9 (PMC12486031: Methods).
| Sample | Paraffin-embedded human colonic adenocarcinoma tissue; fixative not specified (datasheet A06312) |
| Fixation | Image fixative and duration unreported (datasheet A06312); 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 A06312); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06312) |
| Primary antibody | Rabbit anti-CHCHD10, 2-5μg/ml (datasheet A06312) |
| Primary incubation | Overnight at 4 °C (datasheet A06312) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A06312) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CHCHD10-positive staining in endocrine cells of appendix (HPA tissue IHC: High). HPA tissue profile: Granular cytoplasmic expression in several different tissue types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 for the catalog antibody (datasheet A06312); consider the published pH 9 method when adapting the tissue protocol (PMC12486031: Methods).
Section 2
What Is the Expected CHCHD10 Staining Pattern?
CHCHD10 localizes to the mitochondrial intermembrane space and is enriched at cristae junctions; it has no transmembrane segment (UniProt Q8WYQ3). In paraffin-section IHC, expect granular cytoplasmic staining in several tissues, particularly endocrine cells of the appendix and intestine and kidney collecting ducts (HPA: tissue IHC). HPA rates the tissue staining Enhanced, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC).
| Discrete cytoplasmic granules in intestinal or appendix endocrine cells, or kidney collecting ducts. | This fits the reported high-staining cell populations and granular tissue pattern (HPA: tissue IHC). Score the relevant cells and compartment rather than treating every cell in a positive section as positive; expression varies by cell type (HPA: tissue IHC). |
| Predominantly nuclear, surface, or broad extracellular staining, without a granular cytoplasmic component. | This conflicts with mitochondrial localization and the reported tissue pattern (UniProt Q8WYQ3; HPA: tissue IHC). Treat it as a possible staining artefact; general IHC practice: compare morphology, a reagent control, and an independently validated stain before assigning specificity. |
| Strong staining of adipocytes or respiratory epithelial cells instead of the expected positive cells. | HPA reports no detection in adipocytes of adipose tissue or breast, or respiratory epithelium of the bronchus (HPA: tissue IHC). Unexpected staining raises concern for cross-reactivity or endogenous detection activity; general IHC practice: check a no-primary control. It does not establish either cause by itself. |
| Uniform haze across cells, stroma, and empty areas, obscuring discrete granules. | The distribution does not match the reported granular cytoplasmic pattern (HPA: tissue IHC). General IHC practice: assess background with a no-primary control and review blocking, washes, and detection exposure before scoring faint cellular signal. |
| No signal in appendix endocrine cells or kidney collecting ducts. | Both are reported high-staining populations (HPA: tissue IHC), so a blank result calls for a technical check before interpreting the specimen as CHCHD10-negative. General IHC practice: confirm that the expected cells are present and review antibody, retrieval, and detection controls. |
💡Expected CHCHD10 appearanceCall a result positive when distinct granular cytoplasmic staining occurs in expected cells, especially high-staining intestinal or appendix endocrine cells or kidney collecting ducts (HPA: tissue IHC); isolated nuclear or diffuse tissue-wide color is suspicious given mitochondrial localization (UniProt Q8WYQ3; HPA: tissue IHC).
| Cell type and tissue selection | High staining is reported in endocrine cells of the appendix, colon, duodenum, rectum, and small intestine, and in kidney collecting ducts (HPA: tissue IHC). Several other populations have low or undetected staining, so select controls by cell type rather than tissue name alone (HPA: tissue IHC). |
| Antibody validation and interpretation limit | HPA003440 has Enhanced IHC validation; HPA describes medium consistency between antibody staining and RNA expression (HPA: antibody record; tissue IHC). This supports the reported pattern but does not make every stained cell specific. Compare compartment and cell identity when scoring (HPA: tissue IHC; UniProt Q8WYQ3). |
| Mitochondrial position and processing | UniProt places CHCHD10 in the intermembrane space, enriched at cristae junctions, and annotates a mature chain spanning residues 17–142 (UniProt Q8WYQ3). These annotations guide compartment interpretation; they provide no antibody epitope or target-specific retrieval requirement. |
| What should IF/ICC show? | A mitochondrial pattern is supported by HPA ICC-IF images from A-431 and PC-3 cells (HPA: subcellular ICC-IF). This is a separate application: the IHC tissue pattern and ICC-IF localization answer different questions, and the supplied evidence provides no IF/ICC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The positive control is blank. | A technical failure is possible when a reported high-staining population gives no signal (HPA: tissue IHC). Cell composition may also differ within a section. | Verify the expected endocrine cells or collecting ducts are present (HPA: tissue IHC). General IHC practice: check primary antibody, retrieval, detection reagents, and counterstain against a working control. |
| Only nuclei or cell surfaces stain. | The compartment conflicts with mitochondrial localization and granular cytoplasmic tissue staining (UniProt Q8WYQ3; HPA: tissue IHC). The pattern alone cannot identify the source. | General IHC practice: inspect a no-primary control, compare cellular morphology, and repeat with a validated antibody or independent localization readout before calling CHCHD10 positive. |
| Unexpected cells show strong color while expected cells do not. | Cell-type distribution conflicts with HPA observations; for example, bronchial respiratory epithelium is reported undetected while intestinal endocrine cells can be high (HPA: tissue IHC). | Check that cell identities and section landmarks are correct (HPA: tissue IHC). General IHC practice: use a no-primary control to assess endogenous detection activity, then reassess primary-antibody specificity. |
| Granules are hard to distinguish from diffuse background. | Background can obscure the reported granular cytoplasmic pattern (HPA: tissue IHC). Its source cannot be determined from the supplied CHCHD10 evidence. | General IHC practice: compare with a no-primary control, review blocking and washes, and adjust detection exposure before scoring. Keep a clear positive-control section in the same run (HPA: high-staining cell populations). |
| A heart or liver section stains weakly despite expected expression. | UniProt reports higher expression in heart and liver, while the supplied HPA high IHC examples identify other cell populations (UniProt Q8WYQ3; HPA: tissue IHC). Expression level alone does not predict staining intensity in a particular section. | Use the reported high-staining HPA cell populations to assess the IHC run (HPA: tissue IHC). General IHC practice: review section quality and detection controls before concluding the target is absent. |
| The tissue stain seems diffuse, but IF/ICC appears mitochondrial. | HPA reports granular cytoplasmic tissue IHC and supported mitochondrial ICC-IF localization (HPA: tissue IHC; subcellular ICC-IF). The assays present localization at different visual scales. | Score the IHC section against cell identity and granular cytoplasmic appearance (HPA: tissue IHC). Use the ICC-IF result as a localization cross-check, without treating it as proof that diffuse chromogenic staining is specific (HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing CHCHD10 staining
A06312 · IHCIHC analysis of CHCHD10 using anti-CHCHD10 antibody (A06312). CHCHD10 was detected in paraffin-embedded section of human colonic adenocarcin
A06312 · IHCIHC analysis of CHCHD10 using anti-CHCHD10 antibody (A06312). CHCHD10 was detected in paraffin-embedded section of human liver cancer tissue
A06312 · IFIF analysis of CHCHD10 using anti-CHCHD10 antibody (A06312). CHCHD10 was detected in immunocytochemical section of CACO-2 cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000250479-CHCHD10/tissue/cerebral+cortex#imid_1274924
HPA · Small intestineHuman Protein Atlas IHC, Small intestine. Source: https://www.proteinatlas.org/ENSG00000250479-CHCHD10/tissue/small+intestine#imid_1274817
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000250479-CHCHD10/tissue/colon#imid_1274858
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000250479-CHCHD10/tissue/liver#imid_1274859
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003440; green = CHCHD10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA003440; green = CHCHD10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CHCHD10 IHC & IF
🧪Run kidney first and confirm staining in collecting duct cells, which HPA scores High (HPA: kidney collecting ducts, High). Use adipose tissue adipocytes as the negative comparison (HPA: adipocytes, Not detected); on the kidney slide, cells lacking specific signal should have only background staining, but adjacent renal cells are not established negatives because CHCHD10 is ubiquitously expressed (UniProt Q8WYQ3 tissue specificity).
Positive control tissue: Appendix (Endocrine cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHCHD10 in A-431, PC-3, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched nonimmune rabbit IgG control; the selected antibody is rabbit-derived, while its clonality is unreported (A06312 caption: rabbit anti-CHCHD10). Use CHCHD10 knockout tissue if available, or peptide competition if the immunizing peptide is available; for kidney chromogenic staining, quench endogenous peroxidase and check endogenous biotin background when using the caption’s biotin-based SABC/DAB detection (A06312 caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state its fixative (A06312 caption: fixative unreported). The caption uses heat-mediated EDTA retrieval at pH 8.0, but one example does not establish that retrieval is required for every specimen (A06312 caption: EDTA retrieval). Neither frozen sections nor IF/ICC is established as easier by the supplied evidence; IF/ICC can check the supported mitochondrial localization in A-431 or PC-3, while kidney staining with SABC/DAB warrants attention to endogenous biotin background (HPA: mitochondria supported; A06312 caption: SABC/DAB).
HPA tissue IHC evidence for CHCHD10
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 | Endocrine cells | High | Protein (IHC) | HPA → |
| Colon | Endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | Endocrine cells | High | Protein (IHC) | HPA → |
| Kidney | Collecting ducts | High | Protein (IHC) | HPA → |
| Rectum | Endocrine cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CHCHD10 IHC Tips
Troubleshoot CHCHD10 staining in paraffin sections by checking retrieval, cellular pattern, detection controls and cell-specific scoring (datasheet A06312; HPA tissue IHC).
What retrieval should I try first if CHCHD10 staining is weak?
Use heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections stained with A06312 (datasheet A06312). The documented tissue image used this retrieval before applying 2 µg/mL primary antibody overnight at 4°C (datasheet A06312). Keep heating and cooling conditions consistent across comparison sections, since retrieval variation can change chromogenic intensity (standard IHC practice). If staining remains weak, adjust retrieval duration cautiously on matched sections while retaining an identically processed positive control (standard IHC practice). High staining in colon endocrine cells or kidney collecting ducts provides a cell-specific reference pattern (HPA tissue IHC).
Could fixation explain variable CHCHD10 staining between paraffin blocks?
The A06312 tissue caption describes a paraffin section but does not state its fixative (datasheet A06312). CHCHD10-specific sensitivity to fixation is therefore unknown from the supplied evidence (datasheet A06312). Record each block's fixative and fixation duration, then compare sections stained in the same run with EDTA at pH 8.0 and 2 µg/mL primary antibody (standard IHC practice; datasheet A06312). Include a known staining reference section and a no-primary control to distinguish block-dependent signal loss from detection background (standard IHC practice). Do not attribute a weak block to fixation until processing and staining differences have been checked (standard IHC practice).
Where should convincing CHCHD10 signal appear in tissue sections?
Look for granular cytoplasmic staining, the reported tissue pattern, consistent with mitochondrial localisation (HPA tissue IHC; HPA subcellular). CHCHD10 resides in the mitochondrial intermembrane space and is enriched at cristae junctions, so a predominantly nuclear pattern needs scrutiny (UniProt Q8WYQ3 subcellular location). Colon endocrine cells and kidney collecting ducts have reported high staining and can guide cell-level review (HPA tissue IHC). The A06312 caption documents detection in a human colonic adenocarcinoma paraffin section but does not establish which cells stained (datasheet A06312). Compare morphology and compartment across controls before calling broad cytoplasmic colour specific (standard IHC practice).
How should I assess epitope accessibility when the antibody's binding site is unspecified?
The supplied record lists no CHCHD10 isoforms and identifies a processed chain spanning residues 17–142 (UniProt Q8WYQ3 processing and isoforms). Its CHCH domain spans residues 99–140, with no annotated transmembrane segment or glycosylation sites (UniProt Q8WYQ3 domains, topology and glycosylation). The A06312 caption does not identify its binding epitope, so neither domain recognition nor loss of a particular epitope can be inferred (datasheet A06312). Start with the documented EDTA retrieval at pH 8.0, then compare matched sections if accessibility is suspected (datasheet A06312; standard IHC practice). Use an independent antibody or orthogonal expression evidence when resolving discordant staining (standard IHC practice).
How can IF help check a mitochondrial CHCHD10 pattern?
For a separate IF assessment, compare CHCHD10 puncta with a mitochondrial marker and an endocrine-cell marker where those cells are expected (HPA subcellular; HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores, favouring a far-red channel if the tissue shows substantial autofluorescence (standard IF practice). Because CHCHD10 is in the mitochondrial intermembrane space, assess whether permeabilisation exposes the antibody epitope while preserving mitochondrial morphology; the epitope itself is unspecified (UniProt Q8WYQ3 subcellular location; datasheet A06312; standard IF practice). Include single-label and no-primary controls to assess bleed-through and background (standard IF practice). The supplied A06312 paraffin IHC caption does not establish IF fixation or permeabilisation conditions (datasheet A06312).
How can I identify the source of diffuse brown staining?
The documented A06312 section used 10% goat serum blocking, 2 µg/mL primary antibody overnight at 4°C, and a biotin-based DAB detection sequence (datasheet A06312). Compare a no-primary section to identify colour arising from detection reagents or endogenous activity (standard IHC practice). Quench endogenous peroxidase and consider endogenous biotin when using this detection chemistry, particularly if background persists without primary antibody (standard IHC practice; datasheet A06312). If background depends on primary antibody, reassess blocking, primary concentration and washing on matched sections (standard IHC practice). Treat diffuse colour cautiously when it lacks the reported granular cytoplasmic pattern (HPA tissue IHC).
What scoring method best captures cell-specific CHCHD10 staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population before scoring, since reported CHCHD10 staining differs among tissue cell types (HPA tissue IHC). For each region, record the percentage of positive target cells and staining intensity on a 0–3 scale; an H-score ranges from 0–300 (standard IHC scoring practice). Normalise positive counts to the number of evaluable target cells, or report positive-cell density per mm² of evaluable tissue when cell counting is impractical (standard IHC scoring practice). Exclude folds, edges and necrotic areas using the same rules for every section (standard IHC practice). Keep retrieval, detection and image thresholds consistent across compared samples (standard IHC practice).
When should an apparent CHCHD10-positive area be considered artefact?
A convincing result combines granular cytoplasmic staining with an appropriate cell population and low no-primary background (HPA tissue IHC; standard IHC practice). Predominantly nuclear colour conflicts with the supported mitochondrial location and warrants further validation (HPA subcellular; UniProt Q8WYQ3 subcellular location). Strong staining confined to section edges, folds or necrotic areas can reflect processing or detection artefacts (standard IHC practice). Brown signal in a no-primary control suggests endogenous enzyme activity or detection-reagent background rather than CHCHD10-specific binding (standard IHC practice). Interpret unexpected cell patterns cautiously because the HPA tissue profile has only medium consistency with RNA expression data (HPA tissue IHC reliability).
Boster reagents
Best CHCHD10 / Coiled-coil-helix-coiled-coil-helix domain-containing protein 10, mitochondrial IHC Antibodies
A06312 has documented CHCHD10 IHC in human paraffin sections and IF/ICC in CACO-2 cells (catalog image captions).
Real IHC data
Anti-CHCHD10 Antibody ®
Cat # A06312
A06312 is listed for IHC and IF/ICC (catalog applications). Its IHC captions show human paraffin sections of colonic adenocarcinoma, liver cancer, lung cancer, and ovarian serous adenocarcinoma; its IF caption shows CACO-2 cells (catalog image captions).
Which to pick: Choose A06312 for tissue IHC: its own images document human paraffin sections, with 2 μg/ml antibody after EDTA pH 8 retrieval (A06312 IHC captions). Choose A06312 for IF/ICC in CACO-2 cells at 5 μg/ml (A06312 IF caption). The catalog lists human, monkey, mouse, and rat reactivity, but the supplied IHC and IF images establish only the stated samples; clonality and the tissue fixative are unreported (catalog reactivity; A06312 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8WYQ3 (CHC10_HUMAN, Coiled-coil-helix-coiled-coil-helix domain-containing protein 10, mitochondrial).
- Human Protein Atlas. CHCHD10 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CHCHD10 subcellular location (ICC-IF): Localized to the mitochondria..
- Human Protein Atlas. CHCHD10 antibody validation summary (1 antibodies).
- Neuropathologic description of CHCHD10 mutated amyotrophic lateral sclerosis. Neurology. Genetics 2020 — PMC6975173.
- Distinct Muscle Biopsy Findings in Genetically Defined Adult-Onset Motor Neuron Disorders. PloS one 2016 — PMC4801364.
- Modulation of synaptic plasticity, motor unit physiology, and TDP-43 pathology by CHCHD10. Acta neuropathologica communications 2022 — PMC9254494.
- Clinical, neuropathological, and biochemical characterization of ALS in a large CHCHD10 R15L family. medRxiv : the preprint server for health sciences 2025 — PMC12486031.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:24934289 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CHCHD10 (Coiled-coil-helix-coiled-coil-helix domain-containing protein 10, mitochondrial). Expected localisation Granular cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06312). Validated IHC antibody: A06312 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Endocrine cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colonic adenocarcinoma tissue; fixative not specified (datasheet A06312); 2. Fixation: Image fixative and duration unreported (datasheet A06312); 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 A06312); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06312); 8. Primary antibody: Rabbit anti-CHCHD10, 2-5μg/ml (datasheet A06312); 9. Primary incubation: Overnight at 4 °C (datasheet A06312). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); target-specific effects are unknown.; Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: No isoforms listed; processed chain spans residues 17–142 (UniProt); Regulation: Higher expression in heart and liver (UniProt). Evidence: UniProt Q8WYQ3, HPA tissue IHC · Enhanced, HPA ICC-IF · mitochondria., Datasheet A06312, PMC12486031. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CHCHD10/immunohistochemistry-ihc-guide.