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CHDH / Choline dehydrogenase, mitochondrial · IHC design guide
Design Immunohistochemistry for CHDH
Use proximal renal tubules as a high-staining reference for CHDH in paraffin-section IHC (HPA tissue IHC). Assess granular cytoplasmic staining (HPA tissue IHC), with mitochondrial membrane localisation as a molecular expectation (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CHDH 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) | |
| Staining pattern | Granular cytoplasmic staining, strongest in renal tubules (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+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 | Colon stains less strongly than renal tubules (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; mature chain spans aa 30–594 (UniProt) |
Section 1
Recommended CHDH IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with three published CHDH IHC methods (PMC12124313; PMC12421421; PMC9034393).
| Sample | Paraffin-embedded Mouse kidney tissue; fixative not specified (datasheet A10927) |
| Fixation | Image fixative and duration unreported (datasheet A10927); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-CHDH, 1:50-1:200 (datasheet A10927) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CHDH-positive staining in proximal tubules (cell body) of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression with a granular pattern in several tissues, most abundant in renal tubules. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); compare the published retrieval conditions if needed (PMC12124313; PMC9034393).
Section 2
What Is the Expected CHDH Staining Pattern?
CHDH should appear as granular cytoplasmic staining, strongest in kidney proximal tubule cell bodies and also high in testis peritubular cells (HPA tissue IHC: Enhanced). The granular pattern fits its mitochondrial location; UniProt places CHDH at the inner membrane and, after mitochondrial damage, the outer membrane, with no annotated transmembrane segment (UniProt Q8NE62 topology). HPA’s Enhanced tissue reliability reflects consistency between staining and RNA expression (HPA tissue IHC).
| Granular cytoplasmic signal is strong in renal proximal tubule cell bodies. | This is the clearest tissue positive control: HPA reports High staining in these cells and describes the overall pattern as cytoplasmic and granular (HPA tissue IHC). Compare signal within the same section before treating every stained structure as CHDH-positive. |
| Testis peritubular cells stain strongly; glandular cells in appendix, breast, colon, duodenum, or gallbladder stain less strongly. | HPA reports High staining in testis peritubular cells and Medium staining in those glandular cells (HPA tissue IHC). An intensity difference between these listed cell populations can be expected; interpret each population against its own tissue context. |
| A paraffin section shows predominantly nuclear staining, with little granular cytoplasmic signal. | That conflicts with HPA’s tissue IHC profile and warrants an artefact check (HPA tissue IHC: cytoplasmic, granular). HPA separately reports approved nucleolar localization by ICC-IF, so isolated nucleolar fluorescence should be assessed in that application rather than assumed to explain a nuclear chromogenic pattern (HPA subcellular ICC-IF). |
| Strong staining appears in adipocytes, lung alveolar cells, or bone marrow hematopoietic cells. | HPA reports CHDH as Not detected in those cell populations (HPA tissue IHC). Check cell identification and controls; cross-reactivity or endogenous chromogenic activity are possible explanations. A reference result of Not detected is a comparison point, not proof that every specimen must be negative. |
| Color is diffuse across tissue structures, or kidney proximal tubules show no discernible signal. | Diffuse color does not match the HPA granular profile; absent proximal tubule staining misses an HPA High control (HPA tissue IHC). Review background controls, tissue preservation, retrieval and detection performance as general IHC checks before assigning a biological interpretation. |
💡Expected CHDH appearanceCall a result consistent when renal proximal tubule cell bodies show High granular cytoplasmic staining; widespread diffuse color or dominant nuclear staining is suspect in tissue IHC (HPA tissue IHC: Enhanced; UniProt Q8NE62 mitochondrial location).
| Cell population and tissue | HPA reports High proximal tubule and testis peritubular staining, Medium staining in several listed glandular populations, and Not detected staining in listed controls (HPA tissue IHC). Score the identified cells rather than assigning one intensity to an entire organ. |
| Subcellular interpretation | UniProt places CHDH at mitochondrial inner and outer membranes and notes outer membrane accumulation after mitochondrial damage (UniProt Q8NE62). HPA tissue IHC reports a granular cytoplasmic pattern; ordinary chromogenic granules do not resolve individual mitochondrial membranes (HPA tissue IHC; standard IHC interpretation). |
| Antibody evidence | HPA lists HPA036633 and HPA058130 as IHC Enhanced, while HPA036632 has ICC approval without an IHC status in the supplied record (HPA antibodies). Keep validation tied to the application and antibody used; one antibody’s ICC result does not validate another antibody’s paraffin IHC result. |
| Protein processing | UniProt annotates a mature CHDH chain spanning residues 30–594 and no annotated isoforms or glycosylation sites (UniProt Q8NE62). The supplied record gives no antibody epitope, so these annotations cannot predict which retrieval condition will expose it in a paraffin section. |
| IF/ICC localization | For the IF/ICC question, HPA approves mitochondria as the main location and nucleoli as an additional location, with images from CACO-2, Hep-G2 and OE19 (HPA subcellular ICC-IF). This supports an IF localization comparison, not a paraffin IHC protocol choice. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive kidney proximal tubules have no signal. | The result conflicts with HPA High staining in proximal tubule cell bodies; tissue processing, retrieval or detection failure is possible (HPA tissue IHC; general IHC practice). | Confirm the tubules are present, then review the antibody’s IHC-P instructions and run a suitable positive control through the same retrieval and detection workflow (general IHC practice). |
| The entire section develops diffuse chromogen. | Diffuse color obscures the HPA granular cytoplasmic pattern; excess background or endogenous enzyme activity may contribute (HPA tissue IHC; general chromogenic IHC practice). | Inspect a no-primary control, review blocking and washes, and check endogenous activity controls appropriate to the detection chemistry (general IHC practice). |
| Nuclei dominate the chromogenic result. | Predominant nuclear tissue IHC conflicts with HPA’s cytoplasmic granular profile, although HPA separately reports additional nucleolar localization in ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). | Recheck morphology and background controls, then compare an independently IHC-validated antibody if available; do not transfer the ICC-IF call directly to paraffin IHC (HPA antibodies; general IHC practice). |
| Cells listed as Not detected stain as strongly as proximal tubules. | Cell misidentification, cross-reactivity or endogenous chromogenic activity is possible; HPA lists adipocytes, alveolar cells and hematopoietic cells as Not detected (HPA tissue IHC). | Verify the stained cell type against the counterstain and inspect no-primary and detection controls before calling CHDH expression (general IHC practice). |
| Two antibodies give different tissue patterns. | Antibody performance can differ; HPA assigns IHC Enhanced to HPA036633 and HPA058130, while the supplied record gives HPA036632 no IHC status (HPA antibodies). | Check each antibody’s application validation and compare staining in the same HPA High and Not detected cell populations under matched IHC conditions (HPA antibodies; HPA tissue IHC). |
| IF/ICC shows puncta plus nucleolar signal. Is that compatible with CHDH? | Yes: HPA approves mitochondria as the main ICC-IF location and nucleoli as an additional location (HPA subcellular ICC-IF). | Interpret both compartments against IF controls and keep the conclusion specific to ICC-IF; use the HPA granular cytoplasmic profile when judging tissue IHC (HPA subcellular ICC-IF; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing CHDH staining
A10927 · IHCImmunohistochemistry of paraffin-embedded Mouse kidney using CHDH Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrieval with
A10927 · IHCImmunohistochemistry of paraffin-embedded Rat kidney using CHDH Rabbit pAb at dilution of 1:100 .Perform microwave antigen retrieval with 10
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000016391-CHDH/tissue/cerebral+cortex#imid_9831811
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000016391-CHDH/tissue/colon#imid_9831534
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000016391-CHDH/tissue/liver#imid_9831535
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000016391-CHDH/tissue/kidney#imid_9831536
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA036632; green = CHDH, blue = nucleus, red = microtubules.
HPA · ICC-IF CACO-2Human Protein Atlas immunofluorescence, CACO-2, antibody HPA036632; green = CHDH, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CHDH IHC & IF
🧪Run kidney first: proximal tubule cell bodies should stain strongly (HPA: Kidney, proximal tubules, High). Run adipose tissue as a negative comparator because CHDH is not detected in adipocytes (HPA: Adipose tissue, adipocytes, Not detected); cells outside the proximal tubules on the kidney slide can serve as internal background comparators, but their CHDH-negative status is unestablished (HPA: kidney row identifies proximal tubules only).
Positive control tissue: Kidney (Proximal tubules (cell body), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHDH in CACO-2, Hep-G2, OE19, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control; a concentration-matched nonimmune rabbit IgG isotype control for the rabbit polyclonal catalog antibody; and CHDH-knockout tissue as a biological negative if available (selected A10927 caption: Rabbit pAb; standard IHC control practice). For chromogenic kidney IHC, block endogenous peroxidase and check whether endogenous biotin contributes background if using biotin-based detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected A10927 paraffin-section caption leaves the fixative unreported (selected A10927 caption: fixative not stated). Its mouse kidney example uses microwave retrieval in 10 mM PBS, pH 7.2, before staining at 1:100; this provides a starting condition, without establishing that retrieval is required (selected A10927 caption). The evidence does not establish that frozen sections are easier; IF/ICC offers a mitochondrial localization readout, with additional nucleolar localization reported, while kidney chromogenic staining needs background assessment for endogenous peroxidase and, with biotin-based detection, endogenous biotin (HPA subcellular: approved mitochondria and nucleoli; standard IHC practice).
HPA tissue IHC evidence for CHDH
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Kidney | Proximal tubules (cell body) | High | Protein (IHC) | HPA → |
| Testis | Peritubular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | 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 → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CHDH IHC Tips
Troubleshoot CHDH staining in paraffin sections by checking retrieval, cellular distribution, controls and scoring against the expected mitochondrial pattern (UniProt Q8NE62; HPA tissue IHC).
Which retrieval conditions should I try when CHDH staining is weak in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (page retrieval setting: cytoplasmic or membrane antigen). If staining remains weak with catalog antibody A10927, compare a serial section using microwave retrieval in 10 mM PBS, pH 7.2, the condition reported for its mouse kidney paraffin image (A10927 tissue-IHC caption). Hold primary dilution and chromogen development constant across that comparison so retrieval is the variable being assessed (standard IHC practice). Evaluate preservation of tubular morphology alongside the granular cytoplasmic signal expected in kidney, and reject a condition that raises diffuse background without clarifying that pattern (HPA tissue IHC; standard IHC practice).
How should I troubleshoot CHDH staining differences between paraffin blocks with different fixation histories?
CHDH-specific sensitivity to fixative type or fixation duration is unknown from the supplied evidence; the A10927 paraffin-image caption does not state a fixative (A10927 tissue-IHC caption). Record the fixative and processing history for each block, then compare sections from similarly processed blocks with the same retrieval and detection settings (standard IHC practice). Use a kidney control section and check whether proximal tubular cell bodies retain the expected granular cytoplasmic pattern before treating an absent signal as biological (HPA tissue IHC; standard IHC practice). If morphology and staining both vary across blocks, optimise retrieval and primary concentration on adjacent sections while keeping development time constant (standard IHC practice).
What CHDH staining pattern is plausible, and when should compartment staining raise concern?
Expect a granular cytoplasmic pattern in paraffin tissue, particularly in renal proximal tubular cell bodies where staining is reported as high (HPA tissue IHC). CHDH is assigned to mitochondrial inner membrane and, following mitochondrial damage, can accumulate on the outer membrane; it has no annotated transmembrane segment (UniProt Q8NE62 topology and subcellular location). Diffuse nuclear staining alone therefore needs corroboration, although nucleolar localisation is reported in cell imaging and should not be dismissed automatically (HPA subcellular). Compare the suspected signal with a known positive kidney section, a no-primary control and preserved tissue morphology before calling a new compartment positive (HPA tissue IHC; standard IHC practice).
How can I assess an unexpected CHDH pattern when the antibody epitope is unspecified?
The supplied CHDH record lists 0 isoforms and a processed chain spanning residues 30–594, but it does not identify the catalog antibody’s epitope (UniProt Q8NE62 processing and isoforms; A10927 tissue-IHC caption). Do not assign staining differences to an isoform or a particular epitope without antibody-specific evidence (UniProt Q8NE62 isoforms; standard IHC interpretation). Acetylation and succinylation are annotated at several residues, including alternate modifications at 484 and 496, yet their effect on tissue-IHC binding is unreported (UniProt Q8NE62 modified residues). Compare retrieval conditions and controls first; if the pattern persists, assess it with an independently characterised CHDH reagent or orthogonal expression evidence (standard IHC practice).
How should I examine CHDH by IF alongside a cell-type marker?
Treat IF as a separate optimisation while using the paraffin IHC pattern to choose a relevant region: renal proximal tubular cell bodies show high CHDH staining (HPA tissue IHC; standard IF practice). Multiplex CHDH with a validated proximal tubule marker, assigning fluorophores with well-separated emission and choosing a far-red channel where tissue autofluorescence interferes with shorter wavelengths (standard IF practice). Because the antibody epitope and its membrane-facing side are unspecified, titrate permeabilisation and check mitochondrial access without claiming a known epitope orientation (A10927 tissue-IHC caption; UniProt Q8NE62 topology; standard IF practice). Include single-stain and no-primary controls, then assess mitochondrial colocalisation while allowing for reported nucleolar localisation (HPA subcellular; standard IF practice).
What should I adjust if CHDH chromogenic staining becomes diffuse or widespread?
First compare the section with a no-primary control and a positive kidney control, since true tissue staining is described as granular cytoplasmic and abundant in renal tubules (HPA tissue IHC; standard IHC practice). If both controls show diffuse color, review peroxidase blocking, nonspecific binding block, washes and DAB development as general chromogenic workflow variables (standard IHC practice). If only the primary-stained section is diffuse, titrate the antibody around the caption’s 1:100 starting dilution while holding retrieval and development constant (A10927 tissue-IHC caption; standard IHC practice). Also inspect section edges and damaged areas separately from intact cells before assigning a distribution to CHDH (standard IHC practice).
How should I score CHDH staining across kidney sections and compare samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scoring compartment as renal proximal tubular cell bodies and record the fraction of positive cells plus intensity, or calculate an H-score from those measurements (HPA tissue IHC; standard IHC scoring practice). For area-based analysis, report positive cell density per mm² of viable proximal tubular tissue rather than per total section area (HPA tissue IHC; standard IHC scoring practice). Keep retrieval, antibody dilution, chromogen exposure and imaging thresholds fixed across samples, and normalise counts to the number of eligible cells or viable tubule area (standard IHC practice). Report granular cytoplasmic signal separately from diffuse color or nucleolar signal so different compartments do not collapse into one score (HPA tissue IHC; HPA subcellular; standard IHC scoring practice).
How can I distinguish true CHDH positivity from tissue or detection artefact?
A credible positive resembles granular cytoplasmic staining in renal proximal tubular cell bodies, which are reported as strongly stained, rather than color confined to a cut edge or necrotic region (HPA tissue IHC; standard IHC interpretation). Check a no-primary section for endogenous enzyme or detection-system color before attributing a brown deposit to CHDH (standard chromogenic IHC practice). Predominantly nuclear or broadly diffuse staining warrants further validation because the main reported location is mitochondrial, although nucleolar localisation is also reported (UniProt Q8NE62 subcellular location; HPA subcellular). Compare tissue morphology, expected cell distribution and an independently processed positive control before interpreting a weak or unexpected pattern as a biological change (HPA tissue IHC; standard IHC practice).
Boster reagents
Best CHDH / Choline dehydrogenase, mitochondrial IHC Antibodies
A10927 has IHC images of paraffin-embedded mouse and rat kidney and human colon (catalog image captions); no IF/ICC validation is supplied (catalog applications and images).
Real IHC data
Anti-CHDH Antibody
Cat # A10927
A10927 is listed for IHC in human, mouse and rat (catalog applications and reactivity). Its IHC images show paraffin-embedded mouse kidney, rat kidney and human colon stained at 1:100 after microwave retrieval in 10 mM PBS, pH 7.2 (catalog image captions).
Which to pick: Choose A10927 for paraffin-section IHC: its rabbit polyclonal antibody has images for mouse and rat kidney and human colon, with 1:100 staining and microwave retrieval specified in each caption (catalog host and image captions); the fixative is unreported (catalog image captions). For cross-species IHC, A10927 lists human, mouse and rat reactivity (catalog reactivity). No SKU in this payload is validated for IF/ICC, so A10927 cannot be recommended for that application (catalog applications and images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NE62 (CHDH_HUMAN, Choline dehydrogenase, mitochondrial).
- Human Protein Atlas. CHDH tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CHDH subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoli..
- Human Protein Atlas. CHDH antibody validation summary (3 antibodies).
- Prolyl 4-hydroxylase α-subunit family regulation of type I collagen deposition and IL17RB/c-Jun activation synergistically mediate choline dehydrogenase promotion of colorectal cancer metastasis. MedComm 2025 — PMC11702389.
- Mitophagy-Related Gene CHDH Predicts Prognosis and Immune Response and Inhibits Proliferation and Migration in vitro and in vivo of Oral Squamous Cell Carcinoma. Journal of inflammation research 2025 — PMC12124313.
- CHDH Promotes Breast Cancer Metastasis Relying on IL17RB/CREB1 Signalling Activation. Journal of cellular and molecular medicine 2025 — PMC12421421.
- Quantitative proteomic profiling of hepatocellular carcinoma at different serum alpha-fetoprotein level. Translational oncology 2022 — PMC9034393.
- PubMed PMID:16641997 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:24275569 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CHDH (Choline dehydrogenase, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A10927 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Proximal tubules (cell body)); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Mouse kidney tissue; fixative not specified (datasheet A10927); 2. Fixation: Image fixative and duration unreported (datasheet A10927); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CHDH, 1:50-1:200 (datasheet A10927); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Colon stains less strongly than renal tubules (HPA tissue IHC); Isoform / epitope: No isoforms annotated; mature chain spans aa 30–594 (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q8NE62, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A10927, PMC12124313, PMC12421421. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CHDH/immunohistochemistry-ihc-guide.