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SUCLG1 / Succinate--CoA ligase [ADP/GDP-forming] subunit alpha, mitochondrial · IHC design guide
Design Immunohistochemistry for SUCLG1
Plan SUCLG1 staining in paraffin sections around the granular cytoplasmic pattern observed in tissue (HPA tissue IHC). This guide covers fixation, staining controls and interpretation for the IHC-validated antibody.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SUCLG1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Granular cytoplasm (HPA tissue IHC); mitochondrion (UniProt) | |
| Staining pattern | Widespread granular cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06274-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Hippocampus |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Broad staining limits cell-type specificity (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | No isoforms; mature chain spans 41–346 (UniProt) |
Section 1
Recommended SUCLG1 IHC & IF Protocols
The catalog antibody protocol (datasheet A06274-1) and a published paraffin-section protocol (PMC13553469) provide starting conditions for chromogenic SUCLG1 IHC.
| Sample | Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06274-1) |
| Fixation | Image fixative and duration unreported (datasheet A06274-1); 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 A06274-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06274-1) |
| Primary antibody | Rabbit anti-SUCLG1, 2-5 μg/ml (datasheet A06274-1) |
| Primary incubation | Overnight at 4 °C (datasheet A06274-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06274-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SUCLG1-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 for the catalog antibody (datasheet A06274-1). The published protocol used microwave retrieval at pH 9.0 (PMC13553469).
Section 2
What Is the Expected SUCLG1 Staining Pattern?
SUCLG1 is a mitochondrial protein with no transmembrane segment (UniProt P53597). In paraffin-section IHC, expect granular cytoplasmic staining across many cell types (HPA: ubiquitous cytoplasmic expression with a granular pattern). HPA reports high staining in several glandular, endocrine and respiratory epithelial cell populations, and rates the tissue pattern Enhanced for consistency with RNA expression (HPA: tissue IHC).
| Granular cytoplasmic signal in adrenal glandular cells or bronchial respiratory epithelial cells (HPA: High in each). | This fits the reported tissue pattern and mitochondrial location (HPA: tissue IHC; UniProt P53597). Judge signal within the named cells; a uniformly colored section does not establish mitochondrial staining (general IHC practice). |
| Strong, predominantly nuclear staining, with little granular cytoplasmic signal. | A predominantly nuclear pattern conflicts with the mitochondrial location and granular cytoplasmic IHC profile (UniProt P53597; HPA: tissue IHC). Check controls and detection background before interpreting it as SUCLG1 (general IHC practice). |
| Strong signal in hippocampal glial cells, especially if expected positive cells are faint. | HPA reports SUCLG1 as Not detected in those glial cells (HPA: hippocampus IHC). Review cell identification and control staining; the discrepancy may reflect nonspecific binding or endogenous detection activity (general IHC practice). |
| Diffuse color over cells and surrounding tissue, obscuring granules. | This cannot be confidently scored against the reported granular cytoplasmic pattern (HPA: tissue IHC). Background may arise from nonspecific antibody binding or the chromogenic detection system; assess a matched negative control (general IHC practice). |
| No signal in adrenal glandular cells while slide controls appear valid. | That conflicts with HPA's High staining for those cells (HPA: adrenal gland IHC). First verify the cells sampled and the assay controls; a blank result alone cannot establish biological absence (general IHC practice). |
💡Expected SUCLG1 appearanceCall an IHC result positive when the relevant cells show clear granular cytoplasmic staining, including High staining in adrenal glandular cells (HPA: tissue IHC); predominantly nuclear color or diffuse tissue-wide color is discordant with that pattern (HPA: tissue IHC; general IHC practice).
| Compartment and topology (UniProt P53597; HPA: tissue IHC). | SUCLG1 is mitochondrial and lacks a transmembrane segment (UniProt P53597). Interpret the reported granular cytoplasmic IHC pattern as the primary compartment readout (HPA: tissue IHC); do not require a crisp cell-surface rim. |
| Choice of positive and comparator cells (HPA: tissue IHC). | HPA reports High staining in adrenal glandular and bronchial respiratory epithelial cells, Low staining in smooth muscle cells, and Not detected in hippocampal glial cells (HPA: tissue IHC). Compare identified cells, since the reported levels are cell-specific. |
| IHC evidence and antibody validation (HPA: tissue IHC; HPA: antibodies). | The tissue profile has Enhanced reliability for consistency between staining and RNA data (HPA: tissue IHC). HPA036683 and HPA036684 each have Enhanced IHC status (HPA: antibodies); that supports the reported pattern without making every stained cell specific. |
| Processing and epitope uncertainty (UniProt P53597). | UniProt lists a 346-residue precursor and a mature chain at residues 41–346, with no annotated isoforms (UniProt P53597). The supplied record does not locate the antibody epitope, so it cannot predict whether processing changes its IHC readout. |
| Q: What can IF/ICC add? (HPA: subcellular ICC-IF). | A: HPA approves mitochondria as the main IF/ICC location and also reports approved plasma membrane and cytosol locations (HPA: subcellular ICC-IF). Use that as a localisation cross-check; it does not replace the granular cytoplasmic paraffin-IHC criterion (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive cells are blank (HPA: High in adrenal glandular cells). | The sampled cells may differ from the intended positive population, or the IHC run may have failed (general IHC practice). | Confirm cell identity and a positive assay control, then review retrieval, primary-antibody dilution and detection against the validated IHC procedure (general IHC practice). |
| The entire section is evenly colored, including spaces without cells. | Chromogen background or endogenous detection activity may be obscuring the reported granular pattern (general IHC practice; HPA: tissue IHC). | Examine a no-primary control; check blocking and washes, and address endogenous enzyme activity when the chosen detection system requires it (general IHC practice). |
| Signal is mainly nuclear rather than granular cytoplasmic. | The distribution disagrees with mitochondrial SUCLG1 and HPA's tissue-IHC profile (UniProt P53597; HPA: tissue IHC). | Compare with a known positive tissue and no-primary control; review primary-antibody dilution and detection background before scoring (general IHC practice). |
| Hippocampal glial cells stain strongly (HPA: Not detected in these cells). | Cell misidentification, nonspecific binding or endogenous detection activity could explain the mismatch (general IHC practice). | Recheck morphology and negative controls, and compare the staining distribution with a reported High cell population (HPA: tissue IHC; general IHC practice). |
| Smooth muscle cells look weaker than nearby positive cells. | HPA lists smooth muscle cells as Low, so weaker staining can fit the reference profile (HPA: tissue IHC). | Score the named cell populations separately and retain an appropriate positive control; do not interpret weak smooth muscle staining alone as assay failure (HPA: tissue IHC; general IHC practice). |
| A membrane-like accent appears alongside granular cytoplasmic color. | HPA's ICC-IF data include an additional approved plasma membrane location, while tissue IHC reports granular cytoplasm (HPA: subcellular ICC-IF; HPA: tissue IHC). | Base the paraffin-IHC call on the granular cytoplasmic pattern and controls; document the extra distribution without treating IF/ICC localisation as proof of its IHC specificity (general IHC practice; HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing SUCLG1 staining
A06274-1 · IHCIHC analysis of SUCLG1 using anti-SUCLG1 antibody (A06274-1). SUCLG1 was detected in a paraffin-embedded section of human breast cancer tiss
A06274-1 · IHCIHC analysis of SUCLG1 using anti-SUCLG1 antibody (A06274-1). SUCLG1 was detected in a paraffin-embedded section of human lung adenocarcinom
A06274-1 · IFIF analysis of SUCLG1 using anti-SUCLG1 antibody (A06274-1). SUCLG1 was detected in an immunocytochemical section of U20S cells. Enzyme anti
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000163541-SUCLG1/tissue/cerebral+cortex#imid_8885932
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000163541-SUCLG1/tissue/colon#imid_8885850
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000163541-SUCLG1/tissue/liver#imid_8885843
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000163541-SUCLG1/tissue/kidney#imid_8885860
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036683; green = SUCLG1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036683; green = SUCLG1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SUCLG1 IHC & IF
🧪Run breast first and score glandular cells for SUCLG1 staining (HPA: High in breast glandular cells; selected-SKU caption: breast cancer tissue IHC). Use hippocampus glial cells as the biological negative reference (HPA: Not detected in hippocampus glial cells). On the breast slide, cells designated as internal negatives should show only background DAB signal; no specific breast cell type is established as an internal negative by the supplied HPA rows.
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: Hippocampus (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SUCLG1 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit IgG isotype control (selected-SKU caption: rabbit primary antibody; standard IHC practice). Use SUCLG1 knockout material or a validated peptide-block control to test specificity, and quench endogenous peroxidase before DAB detection in breast sections (standard IHC practice; selected-SKU caption: HRP/DAB detection).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected-SKU tissue-IHC caption does not state a fixative (selected-SKU caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used for the paraffin breast cancer section, but retrieval dependence is unreported (selected-SKU caption: EDTA retrieval). HPA shows mitochondrial ICC-IF localization in A-431, U-251MG and U2OS cells; the supplied evidence does not establish that IF or frozen sections are easier, or document a breast-specific artefact (HPA: subcellular ICC-IF images; selected-SKU caption: paraffin breast cancer section).
HPA tissue IHC evidence for SUCLG1
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Endocrine cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cerebellum | White matter cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SUCLG1 IHC Tips
Use the catalog antibody’s paraffin section workflow as a starting point, then judge staining against SUCLG1’s expected mitochondrial pattern and tissue controls.
Which antigen retrieval conditions should I start with for SUCLG1 IHC?
Begin with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06274-1). The selected tissue image used that retrieval before incubation with 2 μg/ml antibody overnight at 4°C (caption A06274-1). If staining is weak, compare retrieval duration on adjacent sections while keeping antibody concentration and chromogen development consistent (standard IHC practice). Check whether the change improves granular cytoplasmic staining in tissue with expected expression, rather than staining every compartment more strongly (HPA tissue IHC; standard IHC practice). Include a section processed without primary antibody to reveal detection-related background (standard IHC practice).
How should I troubleshoot weak staining when fixation history is uncertain?
Target-specific fixation sensitivity for SUCLG1 is unknown from the supplied evidence, and the selected paraffin section caption does not state a fixative (caption A06274-1). Record the fixative and processing history for each specimen, then compare sections with similar handling before changing retrieval or antibody concentration (standard IHC practice). For a reference starting point, the selected image used heat-mediated EDTA at pH 8.0 and 2 μg/ml antibody overnight at 4°C (caption A06274-1). If a poorly stained specimen has uncertain processing, assess a separately processed positive tissue control in the same run (standard IHC practice). Do not assign a fixation cause solely from the staining pattern (standard IHC practice).
What staining pattern is credible for SUCLG1 in paraffin sections?
Look for a granular cytoplasmic pattern consistent with a predominantly mitochondrial protein (HPA tissue IHC; UniProt P53597 subcellular location). HPA reports ubiquitous granular cytoplasmic tissue staining and high staining in breast glandular cells, providing a useful pattern and cell-type comparison (HPA tissue IHC). Its cell imaging also records additional plasma membrane and cytosolic localisation, so an atypical compartment needs confirmation rather than automatic rejection (HPA subcellular). Compare staining with tissue structure and a no-primary control before accepting diffuse signal or sharply stained edges (standard IHC practice). Score cellular staining separately from extracellular deposits and damaged regions (standard IHC practice).
Could processing or protein modifications change which SUCLG1 epitope is detected?
The supplied record lists 0 isoforms and a mature chain spanning residues 41–346, while the protein has no annotated transmembrane segment (UniProt P53597). It also lists acetylation and succinylation at several residues, including alternate modifications at 57 and 66 (UniProt P53597 modified residues). The antibody epitope is not supplied, so these features do not establish whether processing or modification affects this antibody’s tissue staining (supplied record; standard IHC interpretation). If staining differs across specimens, first compare processing and controls under identical EDTA pH 8.0 retrieval conditions (datasheet A06274-1; standard IHC practice). An independent antibody with a documented, distinct epitope could help investigate a persistent discrepancy (standard IHC practice).
How can I assess SUCLG1 by IF alongside this IHC guide?
Treat IF as a separate assay requiring its own antibody and processing validation; the selected catalog image documents paraffin section chromogenic IHC (caption A06274-1). For multiplex IF, pair SUCLG1 with a marker identifying the cell population being evaluated, and assess whether its signal follows the expected mitochondrial distribution within those cells (UniProt P53597 subcellular location; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence and inspect unstained and secondary-only controls before interpreting puncta (standard IF practice). Because SUCLG1 has no annotated transmembrane segment and is mainly mitochondrial, optimise permeabilisation to permit intracellular antibody access without losing structure (UniProt P53597 topology; HPA subcellular; standard IF practice). Do not transfer the IHC caption’s overnight 4°C incubation directly to IF without testing it (caption A06274-1; standard IF practice).
What should I check when SUCLG1 DAB staining looks diffuse?
The selected IHC image used 10% goat serum blocking, a peroxidase-linked secondary antibody, and DAB development (caption A06274-1). Include a no-primary section and apply a peroxidase block to test for detection-system signal, particularly where endogenous enzyme activity could contribute (standard IHC practice). If background persists, review blocking, washes, antibody concentration, and DAB development while changing one variable at a time (standard IHC practice). The image’s 2 μg/ml antibody concentration is a documented starting condition, not proof that it suits every specimen (caption A06274-1; standard IHC practice). Prefer discrete granular cytoplasmic signal over uniform haze when judging SUCLG1 staining (HPA tissue IHC; standard IHC practice).
How should I quantify SUCLG1 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and analysis area before scoring, because HPA reports different staining levels across tissue cell types (HPA tissue IHC; standard IHC practice). An H-score can combine the percentage of cells at each staining intensity, while percent-positive cells or positive-cell density per mm² may suit a simpler question (standard IHC practice). Normalise each result to evaluable tissue area or the number of eligible cells, and apply one threshold across the comparison set (standard IHC practice). Exclude folds, necrosis, and section edges using prespecified criteria, then record whether the scored signal is granular cytoplasmic (HPA tissue IHC; standard IHC practice). Keep retrieval at EDTA pH 8.0 consistent across sections being compared (datasheet A06274-1; standard IHC practice).
How do I distinguish credible SUCLG1 staining from artefact?
A credible result shows granular cytoplasmic staining in appropriate cells, consistent with the tissue profile and predominant mitochondrial localisation (HPA tissue IHC; HPA subcellular). For context, HPA reports high staining in breast glandular cells, low staining in smooth muscle cells, and no detected staining in hippocampal glial cells (HPA tissue IHC). Treat isolated nuclear signal, extracellular deposits, section-edge staining, and staining confined to necrotic areas as reasons to investigate artefact (UniProt P53597 subcellular location; standard IHC practice). Compare a no-primary section and a peroxidase-blocked section when DAB signal could reflect endogenous enzyme activity (standard IHC practice). Additional plasma membrane or cytosolic signal requires supporting controls before interpretation (HPA subcellular; standard IHC practice).
Boster reagents
Best SUCLG1 / Succinate--CoA ligase [ADP/GDP-forming] subunit alpha, mitochondrial IHC Antibodies
A06274-1 has IHC images from paraffin-embedded human breast cancer, lung adenocarcinoma and testicular seminoma sections, plus rat kidney; IF images show U20S cells and human lung cancer sections (catalog image captions).
Real IHC data
Anti-SUCLG1 Antibody ®
Cat # A06274-1
A06274-1 will render with an IHC figure from a paraffin-embedded human breast cancer section; its other IHC captions cover human lung adenocarcinoma, human testicular seminoma and rat kidney (catalog IHC image captions). Its IF captions show U20S cells and a paraffin-embedded human lung cancer section (catalog IF image captions).
Which to pick: Choose A06274-1 for paraffin-section IHC: its caption documents EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml; the fixative is unreported (catalog IHC image caption). Choose the same SKU for IF/ICC because those applications are listed and both cell and tissue IF images are supplied (catalog applications; catalog IF image captions). For human and rat IHC, A06274-1 has images from both species; it is rabbit-hosted, while clonality is unreported (catalog IHC image captions; catalog host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P53597 (SUCA_HUMAN, Succinate--CoA ligase [ADP/GDP-forming] subunit alpha, mitochondrial).
- Human Protein Atlas. SUCLG1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SUCLG1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the plasma membrane and cytosol..
- Human Protein Atlas. SUCLG1 antibody validation summary (2 antibodies).
- Combined machine learning identification and experimental validation of NDUFS8 and SUCLG1 as potential biomarkers for mechanical asphyxia. Frontiers in medicine 2026 — PMC13553469.
- Multi-Omics Integration Identifies a Five-Gene Metabolic Signature With Experimental Validation in Clear Cell Renal Cell Carcinoma. Cancer reports (Hoboken, N.J.) 2026 — PMC13547743.
- PubMed PMID:15815621 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
- PubMed PMID:9128182 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SUCLG1 (Succinate--CoA ligase [ADP/GDP-forming] subunit alpha, mitochondrial). Expected localisation Granular cytoplasm (HPA tissue IHC); mitochondrion (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06274-1). Validated IHC antibody: A06274-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Hippocampus. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A06274-1); 2. Fixation: Image fixative and duration unreported (datasheet A06274-1); 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 A06274-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06274-1); 8. Primary antibody: Rabbit anti-SUCLG1, 2-5 μg/ml (datasheet A06274-1); 9. Primary incubation: Overnight at 4 °C (datasheet A06274-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Broad staining limits cell-type specificity (HPA tissue IHC); Isoform / epitope: No isoforms; mature chain spans 41–346 (UniProt); Regulation: No expression regulation annotated (UniProt). Evidence: UniProt P53597, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A06274-1, PMC13553469. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SUCLG1/immunohistochemistry-ihc-guide.