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SDHA / Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial · IHC design guide
Design Immunohistochemistry for SDHA
Plan chromogenic SDHA IHC around granular cytoplasmic tissue staining (HPA tissue IHC) and mitochondrial inner membrane localisation at the molecular level (UniProt). This guide covers fixation consistency, antibody selection, controls and cell-specific scoring.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SDHA 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 | Tissue: granular cytoplasm (HPA tissue IHC); mitochondrial inner membrane (UniProt) | |
| Staining pattern | Widespread granular cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PA1717) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes have low baseline staining (HPA tissue IHC) | |
| Regulation | RNA enhanced in heart and skeletal muscle (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; mature chain spans residues 43–664; epitope coverage unknown (UniProt) |
Section 1
Recommended SDHA IHC & IF Protocols
The catalog antibody protocol (datasheet PA1717) is followed by four published SDHA IHC protocols (PMC3545041; PMC6563419; PMC7528421; PMC8353944).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PA1717) |
| Fixation | Image fixative and duration unreported (datasheet PA1717); 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 PA1717); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PA1717) |
| Primary antibody | Rabbit anti-SDHA, 2-5 μg/ml (datasheet PA1717) |
| Primary incubation | Overnight at 4 °C (datasheet PA1717) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet PA1717) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SDHA-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression with a granular pattern. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet PA1717). The gastric GIST study used a high-pH retrieval buffer for 25 minutes (PMC3545041).
Section 2
What Is the Expected SDHA Staining Pattern?
SDHA is associated with the mitochondrial inner membrane and has no annotated transmembrane segment (UniProt P31040 topology). In paraffin sections, expect granular cytoplasmic staining across many cell types, including cardiomyocytes, hepatocytes and kidney collecting duct cells (HPA: ubiquitous granular cytoplasmic profile; High in these cells). HPA rates the tissue IHC profile Enhanced, while reporting medium consistency between antibody staining and RNA expression (HPA: reliability description).
| Granular cytoplasmic staining in cardiomyocytes or kidney collecting duct cells. | This fits the reported pattern and High staining in both cell types (HPA: tissue IHC). Granules are consistent with mitochondrial localisation (UniProt P31040 topology; HPA: supported mitochondrial ICC-IF location). Judge intensity against a processed control section. |
| Predominantly nuclear or continuous cell-border staining, with little granular cytoplasmic signal. | That distribution conflicts with the tissue IHC profile (HPA: ubiquitous granular cytoplasmic pattern). HPA also reports a plasma membrane ICC-IF location, but calls it uncertain (HPA: subcellular). Investigate staining specificity before interpreting either pattern as SDHA. |
| Strong staining restricted to adipocytes or chondrocytes while expected positive cells are faint. | HPA reports Low staining in adipocytes and chondrocytes, but High staining in several other cell populations (HPA: tissue IHC). Check for cross-reactivity or endogenous detection activity with appropriate controls; staining in a Low group alone does not prove either cause. |
| Uniform color across cells and surrounding tissue obscures cytoplasmic granules. | Diffuse signal cannot be confidently scored as the granular SDHA pattern (HPA: tissue IHC profile). General IHC practice: compare a no-primary control and review blocking, washes and chromogen development to locate background from the detection workflow. |
| No signal in a heart muscle section expected to be positive. | Cardiomyocytes are reported High (HPA: tissue IHC). First verify that the tissue and detection controls worked. General IHC practice: review antibody dilution and antigen retrieval conditions; a blank section alone cannot establish absent SDHA protein. |
💡Expected SDHA appearanceCall the result positive when High-staining cell types show distinct granular cytoplasmic signal, especially cardiomyocytes or hepatocytes (HPA: tissue IHC); dominant nuclear staining or uniform haze fails that pattern (HPA: granular cytoplasmic profile).
| Compartment and topology | SDHA belongs to the mitochondrial inner membrane without an annotated transmembrane segment (UniProt P31040 topology). Interpret the section-level granular cytoplasmic pattern alongside that location (HPA: tissue IHC); IHC granules do not resolve membrane topology. |
| Tissue and cell selection | Heart muscle cardiomyocytes, liver hepatocytes and kidney collecting ducts are High; adipocytes and chondrocytes are Low (HPA: tissue IHC). Use these reported levels as comparisons, while allowing for the ubiquitous cytoplasmic profile (HPA: tissue IHC). |
| IHC evidence strength | The overall tissue profile is Enhanced, with medium antibody-to-RNA consistency (HPA: tissue IHC reliability). Antibody HPA041981 is IHC Supported; CAB034929 is IHC Enhanced (HPA: antibody validation). These ratings inform confidence, not a guaranteed result in every section. |
| Isoforms and epitope uncertainty | Three isoforms are listed, and the annotated mature chain spans residues 43–664 (UniProt P31040). No antibody epitope is supplied here, so these annotations cannot establish isoform coverage or justify a particular antigen retrieval condition. |
| IF/ICC Q&A: where should signal appear? | Mainly in mitochondria (HPA: supported ICC-IF location). An additional plasma membrane location is uncertain (HPA: subcellular); assess that claim cautiously. IF/ICC staining conditions belong in its separate guide. |
| Situation | Likely cause | Next action |
|---|---|---|
| Heart muscle control is blank. | Cardiomyocytes are reported High (HPA: tissue IHC), so a technical failure is possible; absence alone does not identify its cause. | General IHC practice: check the detection control, antibody dilution and retrieval record, then repeat with a documented positive section. |
| Granules are weak in a nominally High cell population. | The observed intensity falls below the reported level for that specific population (HPA: tissue IHC); section processing or detection may contribute. | General IHC practice: compare sections processed together and review retrieval, antibody incubation and chromogen development. |
| Nuclei dominate the chromogenic signal. | Predominantly nuclear staining differs from the reported granular cytoplasmic profile (HPA: tissue IHC). | General IHC practice: inspect the no-primary control and reassess background and antibody specificity before scoring. |
| Low-staining cells appear stronger than the positive control. | Adipocytes and chondrocytes are reported Low (HPA: tissue IHC); cross-reactivity or endogenous detection activity is possible. | General IHC practice: compare a no-primary control and an established positive tissue before assigning cell-specific SDHA staining. |
| Color is diffuse throughout the section. | Diffuse color obscures the reported granular cytoplasmic pattern (HPA: tissue IHC); detection background is a possible explanation. | General IHC practice: examine no-primary controls and review blocking, washes and chromogen development. |
| Two antibodies give different patterns. | Their IHC validation differs: HPA041981 is Supported and CAB034929 is Enhanced (HPA: antibody validation). That difference does not identify the source of discordance. | Compare each result with the HPA granular cytoplasmic profile and the same positive control (HPA: tissue IHC); review controls and antibody-specific conditions. |
Experiment evidence
IHC & IF images showing SDHA staining
PA1717 · IHCIHC analysis of SDHA using anti-SDHA antibody (PA1717). SDHA was detected in a paraffin-embedded section of human lung cancer tissue. Heat m
PA1717 · IHCIHC analysis of SDHA using anti-SDHA antibody (PA1717). SDHA was detected in a paraffin-embedded section of human breast cancer tissue. Heat
PB9433 · IHCIHC analysis of SDHA using anti-SDHA antibody (PB9433). SDHA was detected in a paraffin-embedded section of human breast cancer tissue. Heat
PB9433 · IHCIHC analysis of SDHA using anti-SDHA antibody (PB9433). SDHA was detected in a paraffin-embedded section of human ovarian cancer tissue. Hea
PB9433 · IFIF analysis of SDHA using anti-SDHA antibody (PB9433). SDHA was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retr
M01753-2 · IHCHuman kidney was stained with Anti-SDHA rabbit antibody
M01753-2 · IHCMouse kidney was stained with Anti-SDHA rabbit antibody
A01753 · IFIF analysis of SDHA using anti-SDHA antibody (A01753). SDHA was detected in immunocytochemical section of Hela cells. Enzyme antigen retriev
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000073578-SDHA/tissue/cerebral+cortex#imid_15284786
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000073578-SDHA/tissue/colon#imid_15284720
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000073578-SDHA/tissue/liver#imid_15284721
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000073578-SDHA/tissue/kidney#imid_15284722
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA041981; green = SDHA, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA041981; green = SDHA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SDHA IHC & IF
🧪Run heart muscle first: cardiomyocytes should stain strongly (HPA: High in cardiomyocytes). HPA detects SDHA in all 45 scored tissues, so there is no validated negative tissue or internal negative cell population on this slide; use no-primary and isotype controls to assess background (HPA: no negative rows).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: SDHA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SDHA in HeLa, MCF-7, U2OS, KOLF2.1J, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control, matching the catalog antibody’s clonality if known (PA1717 caption: rabbit primary; standard IHC practice). Include an SDHA-knockout biological control if available, and block endogenous peroxidase when using HRP detection in heart muscle (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (PA1717 caption). The demonstrated IHC setup uses heat retrieval in EDTA at pH 8.0; whether frozen sections or IF are easier is unestablished by the supplied evidence (PA1717 caption; HPA: mitochondrial ICC-IF localization). Heart muscle may show endogenous peroxidase background with HRP detection (standard IHC practice).
HPA tissue IHC evidence for SDHA
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: SDHA is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced SDHA IHC Tips
Troubleshoot SDHA staining in paraffin sections by checking retrieval, controls, mitochondrial pattern, and scoring before comparing specimens.
How should I adjust retrieval when SDHA staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for this paraffin-section assay (datasheet PA1717). The selected image used that retrieval, 10% goat-serum blocking, and 2 μg/ml primary antibody overnight at 4°C; hold those variables steady while troubleshooting (caption PA1717). If signal remains weak, compare shorter and longer heating intervals on adjacent sections while matching section thickness and detection conditions, and inspect for heat-related tissue damage (standard IHC practice). Judge improvement by granular cytoplasmic staining in a reference such as cardiomyocytes, alongside a no-primary control (HPA tissue IHC; standard IHC practice).
Could fixation explain variable SDHA staining between paraffin blocks?
The selected tissue caption reports paraffin embedding but does not state a fixative, so target-specific fixation sensitivity is unknown (caption PA1717). Record each block’s fixative and fixation time, and compare sections with similar processing histories because fixation differences can alter antigen accessibility and tissue morphology (standard IHC practice). Keep retrieval at EDTA pH 8.0 and the primary concentration at 2 μg/ml while assessing that variation (caption PA1717). If a poorly staining block also has weak reference-cell staining, review processing records and section quality before attributing the result to reduced SDHA expression (standard IHC practice).
What pattern should count as correctly localised SDHA?
Expect a granular cytoplasmic IHC pattern, consistent with SDHA at the mitochondrial inner membrane (HPA tissue IHC; UniProt P31040 subcellular location). SDHA has no annotated transmembrane segment, but that topology does not make diffuse nuclear or uniform membrane-edge staining an expected readout (UniProt P31040 topology; standard IHC interpretation). Compare suspicious staining with a no-primary section and with a reference cell population such as cardiomyocytes, reported as high for SDHA (standard IHC practice; HPA tissue IHC). If apparent plasma-membrane staining persists, document it separately: that additional location is uncertain in HPA subcellular data and needs independent confirmation (HPA subcellular).
Can processing or isoforms change what this antibody detects?
SDHA has 3 annotated isoforms, and its recorded mature chain spans residues 43–664; the supplied evidence does not map this antibody’s epitope (UniProt P31040 isoforms and processing; caption PA1717). Modified residues include FAD-linked histidine 99 and alternatively acetylated or succinylated lysine 179, but their effects on this antibody’s IHC staining are unreported (UniProt P31040 modified residues; caption PA1717). Compare the same antibody across matched sections before assigning a staining difference to an isoform or modification (standard IHC practice). To establish such a claim, obtain epitope information and test appropriately characterised material under the same retrieval and detection conditions (standard IHC practice).
How can IF help assess a chromogenic SDHA pattern?
Use IF as a parallel localisation check for an IHC pattern, pairing SDHA with a validated marker for the expected cell type, such as cardiomyocytes in heart muscle (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and inspect unstained and single-stain controls before calling overlap (standard IF practice). SDHA is associated with the mitochondrial inner membrane and has no annotated transmembrane segment, but the antibody’s epitope orientation is unspecified; optimise permeabilisation to make that epitope accessible while preserving mitochondrial structure (UniProt P31040 location and topology; caption PA1717; standard IF practice). The supplied PA1717 image establishes paraffin-section IHC conditions, not an IF protocol for this antibody (caption PA1717).
How do I reduce diffuse colour or puncta unrelated to SDHA?
First distinguish granular cytoplasmic staining from colour over necrotic areas, section folds, and tissue edges by reviewing morphology and a no-primary control (HPA tissue IHC; standard IHC practice). The selected assay used 10% goat serum, a peroxidase-conjugated secondary, and DAB; apply an appropriate peroxidase block as a general chromogenic IHC step (caption PA1717; standard IHC practice). If background remains high, optimise blocking, washes, antibody concentration, and DAB development one variable at a time while retaining EDTA pH 8.0 retrieval (standard IHC practice; datasheet PA1717). Interpret any improvement against a reference section so background reduction does not mask genuine granular signal (HPA tissue IHC; standard IHC practice).
What is a reproducible way to score SDHA IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and exclude folds, necrosis, and damaged edges before scoring granular cytoplasmic DAB signal (HPA tissue IHC; standard IHC practice). For intensity comparisons, use a prespecified 0–3 intensity scale with percentage-positive cells to calculate an H-score, and report the percentage separately (standard IHC practice). For spatial comparisons, count SDHA-positive cells per mm² of viable tissue or per defined cell compartment, using identical thresholds and imaging settings across sections (standard IHC practice). Normalise each result to the relevant viable cell population and review a reference section and no-primary control across staining runs (standard IHC practice).
When is apparent SDHA loss convincing rather than artefactual?
A convincing positive has granular cytoplasmic staining in viable cells, matching the reported tissue pattern and mitochondrial location of SDHA (HPA tissue IHC; UniProt P31040 subcellular location). Assess a suspected loss within the intended cell population against nearby viable cells and a separately stained reference, since HPA reports both high and low staining across cell types (HPA tissue IHC; standard IHC practice). Treat nuclear-dominant signal, edge accentuation, necrotic deposits, and staining reproduced in the no-primary control as reasons to investigate artefact (standard IHC practice). Check peroxidase blocking when diffuse DAB persists, and avoid assigning biological loss from one compromised section (standard IHC practice).
Boster reagents
Best SDHA / Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial IHC Antibodies
Anti-SDHA options include paraffin-section IHC images from human tissue and mouse heart, kidney IHC images from human, mouse and rat, and an IF image from HeLa cells (catalog image captions).
Real IHC data
Anti-SDHA Antibody ®
Cat # PA1717
Real IHC data
Anti-SDHA Antibody ®
Cat # PB9433
Real IHC data
Anti-SDHA Rabbit Monoclonal Antibody
Cat # M01753-2
Real IF data
Anti-SDHA Antibody ®
Cat # A01753
PA1717 shows human lung cancer IHC, and PB9433 shows human breast cancer IHC; both list human, mouse and rat reactivity (PA1717 and PB9433 image captions; catalog applications/reactivity). M01753-2 shows human, mouse and rat kidney IHC, while A01753 shows HeLa-cell IF/ICC and does not list IHC (M01753-2 image captions; A01753 image caption and applications).
Which to pick: For paraffin-section chromogenic IHC, choose PA1717 or PB9433: each has an IHC image from a paraffin section and a listed 2–5 μg/ml IHC range; the fixative is unreported in both captions (PA1717 and PB9433 image captions; catalog IHC dilutions). For IF/ICC, choose A01753, which lists both applications and has a HeLa-cell IF image; PB9433 also lists IF/ICC (A01753 image caption and applications; PB9433 applications). For IHC across species, consider monoclonal M01753-2 because its catalog includes human, mouse and rat kidney IHC images and lists 1:200–1:1000 for IHC; its image captions do not report section processing or fixative (M01753-2 catalog description, image captions and IHC dilution).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P31040 (SDHA_HUMAN, Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial).
- Human Protein Atlas. SDHA tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SDHA subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the plasma membrane..
- Human Protein Atlas. SDHA antibody validation summary (2 antibodies).
- Immunohistochemical loss of succinate dehydrogenase subunit A (SDHA) in gastrointestinal stromal tumors (GISTs) signals SDHA germline mutation. The American journal of surgical pathology 2013 — PMC3545041.
- Molecular Alterations in Dog Pheochromocytomas and Paragangliomas. Cancers 2019 — PMC6563419.
- Incidence of succinate dehydrogenase and fumarate hydratase-deficient renal cell carcinoma based on immunohistochemical screening with SDHA/SDHB and FH/2SC. Human pathology 2019 — PMC7528421.
- Single nucleotide variants of succinate dehydrogenase A gene in renal cell carcinoma. Cancer science 2021 — PMC8353944.
- PubMed PMID:7798181 — UniProt-cited evidence.
- PubMed PMID:8142412 — UniProt-cited evidence.
- PubMed PMID:10746566 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SDHA (Succinate dehydrogenase [ubiquinone] flavoprotein subunit, mitochondrial). Expected localisation Tissue: granular cytoplasm (HPA tissue IHC); mitochondrial inner membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PA1717). Validated IHC antibody: PA1717 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PA1717); 2. Fixation: Image fixative and duration unreported (datasheet PA1717); 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 PA1717); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PA1717); 8. Primary antibody: Rabbit anti-SDHA, 2-5 μg/ml (datasheet PA1717); 9. Primary incubation: Overnight at 4 °C (datasheet PA1717). Watch for: Fixation: Keep fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Adipocytes have low baseline staining (HPA tissue IHC); Isoform / epitope: 3 isoforms; mature chain spans residues 43–664 ; epitope coverage unknown (UniProt); Regulation: RNA enhanced in heart and skeletal muscle (HPA tissue RNA). Evidence: UniProt P31040, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet PA1717, PMC3545041, PMC6563419. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SDHA/immunohistochemistry-ihc-guide.