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CHDH / Choline dehydrogenase, mitochondrial · IHC design guide
Design Immunohistochemistry for CHDH
Plan CHDH IHC-P around high, granular cytoplasmic staining in renal proximal tubules (HPA tissue IHC). Start the IHC-validated antibody at 1:50–1:200 (datasheet) and include adipocytes as a comparison reported as undetected (HPA tissue IHC).
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 in tissue (HPA tissue IHC) | |
| Staining pattern | Proximal renal tubules: granular cytoplasmic staining (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 | Mitochondrial damage can shift CHDH to the outer membrane (UniProt) | |
| Regulation | Expression regulation not annotated (UniProt) | |
| Isoform / epitope | No isoforms annotated; processed chain spans 30–594 (UniProt) |
Section 1
Recommended CHDH IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 3 published CHDH IHC protocols (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 staining needs optimization.
Section 2
What Is the Expected CHDH Staining Pattern?
CHDH is a mitochondrial membrane protein with no annotated transmembrane segment (UniProt Q8NE62 topology). In paraffin-section IHC, expect granular cytoplasmic staining, strongest in kidney proximal tubule cell bodies and testis peritubular cells (HPA tissue IHC: High). HPA rates the tissue staining profile Enhanced, citing consistency between antibody staining and RNA expression (HPA tissue IHC: Enhanced).
| Granular cytoplasmic staining is strong in kidney proximal tubule cell bodies (HPA tissue IHC: High). | This fits the reported tissue pattern and mitochondrial location (HPA tissue IHC; UniProt Q8NE62 localization). Evaluate the tubule cells themselves: staining elsewhere in the section does not establish the expected positive result. |
| Peritubular cells stain strongly in testis; glandular cells stain moderately in colon or duodenum (HPA tissue IHC: High; Medium). | These are supported cell-specific positive patterns (HPA tissue IHC). Compare the signal with the expected cell population before scoring the whole tissue as positive. |
| The dominant signal is nuclear, membranous at the cell surface, or uniformly fills cytoplasm without granules. | That differs from the reported granular tissue pattern (HPA tissue IHC) and mitochondrial localization (UniProt Q8NE62). Treat it as suspect IHC staining; HPA also reports nucleolar signal in ICC-IF (HPA subcellular), so nuclear observations need application-specific review. |
| Strong chromogen appears in adipocytes or lung alveolar cells (HPA tissue IHC: Not detected). | These are poor CHDH-positive controls under the reported IHC profile (HPA tissue IHC). Check cell identity, nonspecific antibody binding, and endogenous chromogenic activity (standard IHC practice) before calling the staining CHDH. |
| Kidney proximal tubule cell bodies have no detectable signal (HPA tissue IHC: High). | An expected positive population is missing (HPA tissue IHC). First assess the section and assay controls, including retrieval and detection (standard IHC practice); a negative slide alone cannot establish absence of CHDH. |
💡Expected CHDH appearanceCall positive when strong, granular cytoplasmic signal marks kidney proximal tubule cell bodies (HPA tissue IHC: High; granular pattern); dominant nuclear or uniform haze is suspect in IHC (HPA tissue IHC: granular pattern).
| Tissue and cell selection | Kidney proximal tubule cell bodies and testis peritubular cells are reported High; several glandular populations are Medium (HPA tissue IHC). Choose a positive control by cell population, not by organ name alone. |
| Subcellular pattern | CHDH localizes to mitochondrial inner and outer membranes; outer-membrane accumulation follows mitochondrial damage (UniProt Q8NE62 localization). Routine IHC should be read against the reported granular cytoplasmic pattern (HPA tissue IHC), without inferring membrane-side resolution. |
| Antibody-specific validation | HPA lists IHC Enhanced for HPA036633 and HPA058130, but no IHC status for HPA036632 (HPA antibodies). Match the interpretation to the antibody used; HPA's tissue-level Enhanced rating does not validate every antibody. |
| Maturation and epitope | The annotated mature chain spans residues 30–594, with no annotated transmembrane segment or isoform (UniProt Q8NE62 processing; topology; isoforms). Epitope location for the catalog antibody is not supplied, so these annotations cannot predict its retrieval requirement. |
| Chromogenic detection | Endogenous enzyme activity can produce color in enzyme-based IHC detection (standard IHC practice). A detection-control section helps distinguish such color from antibody-dependent signal; HPA staining levels alone cannot resolve that cause. |
| IF/ICC Q: Should the IHC pattern match IF exactly? | A: Expect mitochondrial localization in ICC-IF; HPA additionally approves nucleolar localization (HPA subcellular). Those ICC-IF observations inform compartment review but do not supply an IHC protocol or establish nucleolar staining in tissue IHC. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in kidney proximal tubules (HPA tissue IHC: High). | The positive population may be absent from the examined field, or retrieval, primary incubation, or detection may have failed (standard IHC practice). | Confirm proximal tubule cell bodies are present, then review the positive control and assay steps (standard IHC practice). Optimize retrieval only as an assay variable; CHDH-specific fixation sensitivity is unreported here. |
| Granules are obscured by widespread cytoplasmic haze (HPA tissue IHC: granular pattern). | Nonspecific antibody or detection background can reduce pattern contrast (standard IHC practice). | Compare with a primary-antibody omission control; review blocking, washes, antibody concentration, and detection exposure (standard IHC practice). Score CHDH only where cellular granularity remains interpretable. |
| Dominant nuclear IHC staining replaces cytoplasmic granules (HPA tissue IHC: granular pattern). | The tissue pattern conflicts with the reported IHC appearance (HPA tissue IHC); the approved nucleolar observation comes from ICC-IF (HPA subcellular). | Check the antibody identity and controls, and reassess the compartment in a known-positive tissue (standard IHC practice). Do not transfer the ICC-IF nucleolar call to tissue IHC without supporting tissue evidence. |
| Adipocytes or lung alveolar cells stain strongly (HPA tissue IHC: Not detected). | Nonspecific binding or chromogenic detection activity is possible (standard IHC practice); these cell populations are reported as not detected (HPA tissue IHC). | Verify cell identity and compare primary-antibody omission and detection controls (standard IHC practice). Treat staining as unresolved until it can be distinguished from background. |
| Color appears broadly, including outside the expected cell populations. | Endogenous detection activity or excessive assay background may contribute (standard IHC practice); CHDH's reported IHC distribution is cell-specific (HPA tissue IHC). | Inspect the detection-control section, blocking, washes, and chromogen development (standard IHC practice). Reassess positives by cell type and granular pattern (HPA tissue IHC). |
| An antibody yields a pattern unlike the HPA tissue profile. | IHC validation differs by antibody: two listed antibodies are Enhanced, while one has no IHC status (HPA antibodies). | Check which antibody produced the slide and review its IHC validation before interpreting discordance (HPA antibodies). Use the reported positive cell populations as comparison points (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 and score proximal tubule cell bodies for staining (HPA: High in kidney proximal tubules). Use adipose tissue adipocytes as the negative comparator (HPA: Not detected in adipocytes); on the kidney slide, record morphologically unstained cells outside proximal tubules as internal negative cells without assuming every other renal cell type is CHDH-negative.
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 no-primary/secondary-only and concentration-matched nonimmune rabbit IgG controls for the rabbit polyclonal antibody (A10927 caption: Rabbit pAb), plus CHDH knockout tissue if available as a biological negative. In kidney, quench endogenous peroxidase and check for endogenous biotin if using biotin-based detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected mouse-kidney paraffin caption does not state a fixative (A10927 caption). That caption uses microwave retrieval in 10 mM PBS at pH 7.2 and a 1:100 dilution, but does not establish that retrieval is required (A10927 caption). The supplied evidence does not establish whether frozen sections or IF are easier; ICC-IF images are available for CACO-2, Hep-G2 and OE19 (HPA subcellular), while renal endogenous biotin can complicate biotin-based IHC (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 pattern, controls and scoring against the reported IHC evidence.
Which retrieval conditions should I try first for CHDH in paraffin sections?
Start with citrate buffer at pH 6.0, heated to 95–98 °C for 20 min (this page’s antigen retrieval setting). Let sections cool consistently before blocking, then compare staining in renal tubules across slides processed together (standard IHC practice; HPA: high in kidney proximal tubules). If signal is weak, test the reported microwave retrieval in 10 mM PBS at pH 7.2 on a separate section; that condition accompanied a 1:100 stain of paraffin-embedded mouse kidney with A10927 (A10927 tissue-IHC caption). Keep detection and exposure to chromogen constant when comparing retrieval conditions (standard IHC practice).
Could fixation explain weak or uneven CHDH staining?
CHDH-specific sensitivity to fixation is unknown because no target-specific fixation condition or comparison is supplied (A10927 tissue-IHC caption: fixative not stated). The mouse kidney image documents paraffin embedding and retrieval, but its unstated fixative cannot establish an optimal fixation time (A10927 tissue-IHC caption). Check tissue preservation, section adhesion and whether matched samples underwent comparable fixation before changing retrieval or antibody concentration (standard IHC practice). Compare suspect sections with a consistently processed positive-control section containing renal tubules, and record any fixation differences when interpreting weak staining (standard IHC practice; HPA: high in kidney proximal tubules).
What staining pattern is plausible for CHDH in renal tissue?
Look for granular cytoplasmic staining in renal tubular cell bodies, especially proximal tubules, as the tissue reference pattern (HPA tissue IHC: granular cytoplasmic pattern; high in kidney proximal tubules). CHDH is associated with mitochondrial inner and outer membranes, with outer-membrane accumulation reported after mitochondrial damage (UniProt Q8NE62: subcellular location). At routine chromogenic IHC resolution, assess a punctate or granular cytoplasmic pattern rather than claiming that individual mitochondrial membranes are resolved (standard IHC practice; UniProt Q8NE62: mitochondrial location). Strong, uniform nuclear chromogen or diffuse staining unrelated to tubules warrants review of morphology and controls (standard IHC practice; HPA tissue IHC: renal tubular pattern).
How should CHDH processing and epitope uncertainty affect antibody selection?
CHDH is listed as a 594-residue precursor with a processed chain spanning residues 30–594, and no isoforms are annotated (UniProt Q8NE62: processing and isoforms). Its listed topology has no transmembrane segment, although mitochondrial membrane localisation is reported; membrane assignment alone therefore does not establish which side contains the antibody epitope (UniProt Q8NE62: topology and subcellular location). Check the catalog antibody’s stated immunogen or epitope against the processed chain before interpreting an absent IHC signal (standard IHC practice; UniProt Q8NE62: processing). Acetylation and succinylation are annotated at selected residues, including alternate modifications at 484 and 496; their effect on this antibody’s staining is unknown (UniProt Q8NE62: modified residues).
How can IF help evaluate an ambiguous chromogenic CHDH pattern?
Use IF as a separate check of the IHC pattern, pairing CHDH with a marker that identifies the expected renal tubular cell population and assessing both signals within the same cells (HPA tissue IHC: high in kidney proximal tubules; standard IF practice). Include single-label controls and choose a fluorophore channel with low tissue autofluorescence so apparent puncta are distinguishable from background (standard IF practice). Because the antibody epitope’s membrane-facing side is unspecified, compare a mild permeabilisation condition with an appropriate control before treating absent mitochondrial signal as biological (UniProt Q8NE62: mitochondrial membrane location and no transmembrane segment; standard IF practice). HPA reports mainly mitochondrial IF localisation and additional nucleolar localisation, so assess both observations in context (HPA subcellular).
What should I check when CHDH DAB staining appears widespread?
Compare the section with a no-primary control and inspect pigment, precipitate and staining at tissue edges before raising the CHDH score (standard chromogenic IHC practice). Apply a peroxidase block before DAB detection and verify that the control remains clean; these are general workflow checks, not CHDH-specific validation (standard chromogenic IHC practice). Review blocking, washes and primary-antibody concentration alongside the reported 1:100 A10927 mouse-kidney image condition, without assuming that dilution transfers unchanged to every specimen (A10927 tissue-IHC caption; standard IHC practice). A convincing pattern should retain granular cytoplasmic contrast in renal tubules rather than coat unrelated structures uniformly (HPA tissue IHC: granular pattern, high in kidney proximal tubules).
How should I score CHDH staining across comparable sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring; renal proximal tubules provide a documented high-staining reference (HPA tissue IHC: high in kidney proximal tubules; standard IHC practice). Report the percentage of positive tubular cells and an intensity-based H-score, using the same scoring thresholds for all slides (standard IHC practice). If measuring positive area or cell density, normalise to evaluable tubular area in mm² or to the number of eligible tubular cells, respectively (standard image-analysis practice). Exclude damaged edges and poorly preserved tissue, and keep retrieval, DAB development and image settings consistent across the comparison (standard IHC practice).
When does a positive CHDH stain look credible rather than artifactual?
A credible kidney result shows granular cytoplasmic staining in proximal tubular cell bodies, supported by an appropriately processed control (HPA tissue IHC: high in kidney proximal tubules and granular cytoplasmic pattern; standard IHC practice). The pattern is compatible with CHDH’s mitochondrial location, although DAB staining alone cannot establish whether signal is on an inner or outer membrane (UniProt Q8NE62: subcellular location; standard IHC practice). Recheck staining confined to cut edges, necrotic areas or unexpected compartments against tissue morphology and a no-primary control (standard chromogenic IHC practice). Persistent chromogen in that control calls for review of endogenous enzyme blocking and detection conditions before assigning CHDH positivity (standard chromogenic IHC practice).
Boster reagents
Best CHDH / Choline dehydrogenase, mitochondrial IHC Antibodies
A10927 has real IHC images of paraffin-embedded mouse and rat kidney and human colon (catalog image captions). No IF/ICC data are supplied (catalog applications and image captions).
Real IHC data
Anti-CHDH Antibody
Cat # A10927
A10927 lists IHC use and human, mouse and rat reactivity (catalog applications/reactivity). Its IHC images show mouse kidney, rat kidney and human colon at 1:100 (catalog image captions).
Which to pick: Choose A10927 for paraffin-section tissue IHC: it is a rabbit polyclonal antibody with an IHC range of 1:50–1:200 (catalog image captions; catalog host and dilution). It is also the cross-species choice here, with human, mouse and rat reactivity listed and IHC images from all three species (catalog reactivity and image captions). No IF/ICC antibody is supported by this payload (catalog applications and image captions); the A10927 captions report paraffin embedding but do not report a fixative (catalog image captions).
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 in tissue (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: Mitochondrial damage can shift CHDH to the outer membrane (UniProt); Isoform / epitope: No isoforms annotated; processed chain spans 30–594 (UniProt); Regulation: Expression regulation not 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.