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OMA1 / Metalloendopeptidase OMA1, mitochondrial · IHC design guide
Design Immunohistochemistry for OMA1
Plan OMA1 staining in paraffin sections using kidney tubules as a high-staining reference (HPA tissue IHC). This guide covers fixation consistency, antigen retrieval, chromogenic detection and interpretation of the cytoplasmic tissue pattern (HPA tissue IHC; datasheet A06260).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
OMA1 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 | Cytoplasmic staining (HPA tissue IHC); mitochondrial inner membrane (UniProt) | |
| Staining pattern | Cytoplasmic staining in several tissues (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06260) | |
| 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 | Skeletal myocytes undetected despite reported strong expression (HPA tissue IHC; UniProt) | |
| Regulation | Stress activates OMA1; staining effect unknown (UniProt) | |
| Isoform / epitope | 2 isoforms; precursor processing may affect epitopes (UniProt) |
Section 1
Recommended OMA1 IHC & IF Protocols
The catalog antibody specifies heat retrieval with EDTA pH 8.0 (datasheet A06260). The published OMA1 IHC protocols below used heart tissue and osteosarcoma specimens (PMC6038665; PMC10102293).
| Sample | Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A06260) |
| Fixation | Image fixative and duration unreported (datasheet A06260); 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 A06260); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06260) |
| Primary antibody | Rabbit anti-OMA1, 2-5 μg/ml (datasheet A06260) |
| Primary incubation | Overnight at 4 °C (datasheet A06260) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06260) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | OMA1-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A06260). The heart study reports retrieval without conditions (PMC6038665).
Section 2
What Is the Expected OMA1 Staining Pattern?
OMA1 is an inner mitochondrial membrane protein with a transmembrane segment at residues 196–216 (UniProt Q96E52 topology). In paraffin tissue sections, expect predominantly cytoplasmic staining, strongest in kidney tubule cells and moderate in several epithelial and hematopoietic populations (HPA tissue IHC). HPA rates its tissue staining Approved, with medium consistency between antibody staining and RNA expression; interpret intensity in that context (HPA tissue IHC).
| Kidney tubule cells show strong cytoplasmic chromogen, while nearby cells vary in intensity. | This fits the reported High tubule-cell signal and broader cytoplasmic tissue profile (HPA tissue IHC). The visible cytoplasmic pattern is compatible with mitochondrial OMA1, although chromogenic IHC alone cannot resolve the inner membrane (UniProt Q96E52 localization; standard IHC practice). |
| The apparent positive result is confined to nuclei, with little cytoplasmic staining in a tissue section. | Recheck specificity and morphology: tissue IHC is described as cytoplasmic (HPA tissue IHC). Nuclear staining is not automatically impossible, since HPA also reports approved nucleoplasmic localization in ICC-IF; that observation should not be treated as a tissue IHC pattern (HPA subcellular ICC-IF). |
| Strong staining appears in adipocytes or skeletal-muscle myocytes. | These cell types were Not detected in HPA tissue IHC, so assess cross-reactivity and endogenous detection activity before scoring them positive (HPA tissue IHC; standard IHC practice). Skeletal muscle deserves particular caution because UniProt reports strong tissue expression, creating a source-level discrepancy rather than proof that either result is universal (UniProt Q96E52 tissue specificity). |
| Color spreads evenly across tissue and empty spaces, obscuring cell boundaries. | That distribution does not resemble HPA's cellular, cytoplasmic profile (HPA tissue IHC). Review background controls, blocking, washes and chromogen development as general IHC checks; the supplied sources do not identify an OMA1-specific cause (standard IHC practice). |
| Kidney tubule cells show no convincing signal while the slide's controls are readable. | A negative result conflicts with the reported High tubule-cell staining (HPA tissue IHC). Check staining-run performance and antibody suitability before inferring absence of OMA1; HPA's Approved rating has only medium staining-to-RNA consistency (HPA tissue IHC; standard IHC practice). |
💡Expected OMA1 appearanceCall a paraffin-section result consistent when kidney tubule cells show High, predominantly cytoplasmic staining; nuclear-only staining, uniform haze or strong adipocyte staining needs review against the HPA tissue pattern (HPA tissue IHC).
| Compartment and topology | OMA1 is assigned to the mitochondrial inner membrane, with matrix-facing residues 144–195, a membrane segment at 196–216 and an intermembrane-space region from residue 217 onward (UniProt Q96E52 topology). This supports a mitochondrial interpretation of cytoplasmic signal but does not establish where the antibody binds. |
| Cell-specific reference staining | HPA reports High staining in kidney tubule cells and Medium staining in appendix, colon and duodenum glandular cells, among other listed populations (HPA tissue IHC). Compare like cell types when assessing intensity; a whole-organ label can hide differences between cells. |
| Negative-reference limits | HPA lists adipocytes, skeletal-muscle myocytes and several other specified cell types as Not detected (HPA tissue IHC). These are observations for named cells, not a claim that every cell in those organs lacks OMA1. UniProt separately reports strong skeletal-muscle expression (UniProt Q96E52 tissue specificity). |
| Antibody-validation scope | The listed antibody, HPA055120, is Approved for IHC and ICC, while the overall tissue profile has medium staining-to-RNA consistency (HPA antibodies; HPA tissue IHC). The supplied record does not report an Enhanced IHC validation, so an unexpected pattern needs independent review. |
| Processing and isoforms | UniProt records 2 isoforms and a chain beginning at residue 144, with a propeptide at residues 14–143 (UniProt Q96E52 processing and isoforms). The supplied sources do not locate the antibody epitope or show how these forms alter tissue staining; do not assign a specific staining pattern to either form. |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubules are blank or much weaker than expected. | The result disagrees with HPA's High tubule-cell reference, but the source does not identify a target-specific failure mechanism (HPA tissue IHC). | Verify the positive-control section, detection reagents and staining run; then review the antibody's documented IHC conditions (standard IHC practice). Treat any retrieval adjustment as a general optimization, not a proven OMA1 fixation effect. |
| Most tissue compartments show equally strong color. | An indiscriminate pattern is difficult to reconcile with HPA's cell-dependent levels and cytoplasmic profile (HPA tissue IHC). General background or detection activity may contribute (standard IHC practice). | Compare a no-primary control, inspect tissue edges and empty areas, and review blocking, washes and development time (standard IHC practice). Re-score only identifiable cells after background is controlled. |
| The stain is nuclear-only in paraffin tissue. | HPA tissue IHC describes cytoplasmic expression, whereas approved nucleoplasmic localization comes from ICC-IF images (HPA tissue IHC; HPA subcellular ICC-IF). | Check the hematoxylin counterstain and cell boundaries, then compare the IHC pattern with a suitable control (standard IHC practice). Do not transfer an ICC-IF compartment call directly to paraffin-section scoring. |
| Adipocytes or skeletal-muscle myocytes stain strongly. | Both are Not detected in HPA's named-cell tissue observations; UniProt's strong skeletal-muscle expression makes the muscle result especially source-dependent (HPA tissue IHC; UniProt Q96E52 tissue specificity). | Confirm cell identity, examine no-primary background and seek independent specificity evidence before accepting the signal (standard IHC practice). Record the HPA–UniProt discrepancy when interpreting skeletal muscle. |
| Moderate staining in glandular cells is called negative because it is weaker than kidney. | HPA reports Medium glandular-cell staining in appendix, colon and duodenum, versus High staining in kidney tubule cells (HPA tissue IHC). | Score intensity within the identified cell population and compare with its tissue reference under the same staining run (HPA tissue IHC; standard IHC practice). Preserve the distinction between moderate signal and absence. |
| Does an IF/ICC image showing nuclear signal invalidate cytoplasmic IHC? | HPA approves both nucleoplasmic and mitochondrial ICC-IF localization, while its tissue IHC summary says cytoplasmic expression (HPA subcellular ICC-IF; HPA tissue IHC). | Interpret each application against its own HPA observations. For paraffin IHC, assess the cell-specific cytoplasmic pattern; use the separate IF/ICC guide for fluorescence interpretation (HPA tissue IHC; HPA subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing OMA1 staining
A06260 · IHCIHC analysis of OMA1 using anti-OMA1 antibody (A06260). OMA1 was detected in a paraffin-embedded section of human colon cancer tissue. Heat
A06260 · IHCIHC analysis of OMA1 using anti-OMA1 antibody (A06260). OMA1 was detected in a paraffin-embedded section of human ovarian cancer tissue. Hea
A06260 · IFIF analysis of OMA1 using anti-OMA1 antibody (A06260) and anti-Tubulin Alpha antibody (M03989-3). OMA1 was detected in immunocytochemical se
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000162600-OMA1/tissue/cerebral+cortex#imid_15680687
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000162600-OMA1/tissue/colon#imid_15680621
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000162600-OMA1/tissue/liver#imid_15680622
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000162600-OMA1/tissue/kidney#imid_15680607
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA055120; green = OMA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA055120; green = OMA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for OMA1 IHC & IF
🧪Run kidney first and expect staining in tubular cells (HPA: kidney cells in tubules, High). Use adipose tissue as a negative comparator (HPA: adipocytes, Not detected); on the kidney slide, assess neighboring cells without specific staining as internal background references without assuming they are OMA1-null (standard IHC practice).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show OMA1 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Mitochondria (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host and clonality, and OMA1 knockout material or an immunizing-peptide block if available (standard IHC practice; A06260 caption: rabbit primary). For kidney sections, quench endogenous peroxidase before HRP/DAB detection and block endogenous biotin if using avidin-biotin detection, since renal tubules can confound those readouts (standard IHC practice; A06260 caption: HRP/DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (A06260 caption). That example uses heat retrieval in EDTA at pH 8.0 before 2 μg/ml primary antibody overnight at 4°C, but it does not establish that retrieval is required (A06260 caption). The supplied evidence does not establish whether frozen sections or IF are easier; kidney tubules may show endogenous peroxidase or biotin artefacts with the corresponding detection methods (standard IHC practice).
HPA tissue IHC evidence for OMA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Colon | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced OMA1 IHC Tips
Troubleshoot OMA1 staining in paraffin sections by checking retrieval, compartment, tissue controls, and scoring before interpreting chromogenic signal.
How should I retrieve OMA1 in paraffin sections when staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A06260). The selected OMA1 tissue image used this retrieval before 2 μg/ml primary antibody overnight at 4°C, so keep those conditions fixed while checking whether retrieval restores signal (datasheet A06260). Compare a retrieved section with a section processed without retrieval, using kidney tubules as a positive tissue reference because their reported staining is high (HPA: High in kidney tubule cells; general IHC practice). If signal remains weak, adjust heating time on matched sections and inspect tissue preservation before considering another buffer as a fallback (general IHC practice).
Can fixation explain weak or variable OMA1 staining?
OMA1-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet A06260). Record the fixative, fixation duration, and processing history for each block, then compare sections processed together to limit variation unrelated to the antibody (general IHC practice). Keep EDTA retrieval at pH 8.0 and the primary concentration at 2 μg/ml while assessing whether staining changes across blocks with different fixation histories (datasheet A06260; general IHC practice). Check morphology alongside signal: damaged or detached tissue makes a fixation or retrieval comparison difficult to interpret, even when DAB is visible (general IHC practice).
Where should convincing OMA1 staining appear in tissue sections?
Expect a predominantly cytoplasmic staining pattern at light-microscope resolution, consistent with the reported tissue IHC profile and OMA1’s mitochondrial inner-membrane location (HPA: cytoplasmic expression in several tissues; UniProt Q96E52 localisation). Compare staining within the relevant cell population: kidney tubule cells are reported as high, while colon glandular cells are reported as medium (HPA: kidney and colon tissue IHC). Diffuse nuclear-only DAB should prompt a specificity check, although approved cell-based imaging also reports nucleoplasmic localisation (HPA: subcellular localisation). Use matched negative controls and assess whether the apparent pattern tracks intact cells rather than tissue folds or section edges (general IHC practice).
Could epitope position or OMA1 processing change the IHC pattern?
OMA1 has 2 reported isoforms, a transmembrane segment at residues 196–216, and annotated processing, so epitope identity matters when comparing antibodies (UniProt Q96E52 isoforms, topology, processing). The record places residues 144–195 on the matrix side and residues from 217 on the intermembrane-space side; check the antibody’s stated immunogen before assigning either side to its epitope (UniProt Q96E52 topology; general IHC practice). A change in DAB intensity alone cannot establish which isoform or processed form was detected (general IHC practice). If the epitope is unspecified, report staining as antibody-detected OMA1 immunoreactivity and assess specificity with an independent control (general IHC practice).
How can I check OMA1 localisation by IF alongside the IHC result?
On the separate IF/ICC workflow, multiplex OMA1 with a marker of the cell population being assessed and a mitochondrial marker to examine cellular identity and spatial overlap (general IF practice; UniProt Q96E52 localisation). Choose fluorophores after viewing an unstained tissue section, placing the weaker signal in a spectral channel with less tissue autofluorescence (general IF practice). Permeabilisation needs to admit antibody to the relevant side of the mitochondrial inner membrane, but the antibody’s epitope must be identified before choosing conditions for matrix-side versus intermembrane-space-side access (UniProt Q96E52 topology; general IF practice). Treat apparent nuclear overlap cautiously because cell-based imaging reports both nucleoplasmic and mitochondrial locations (HPA: subcellular localisation).
What should I change when OMA1 DAB staining is diffuse?
First compare a no-primary control and inspect section edges, folds, and damaged areas to distinguish background from cell-associated signal (general IHC practice). The selected tissue image used 10% goat serum blocking, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary followed by DAB (datasheet A06260). Confirm that the general chromogenic workflow includes an endogenous-peroxidase block and adequate washes, then titrate primary antibody downward if the no-primary control is clean but diffuse staining persists (general IHC practice). Judge any improvement against intact kidney tubule cells, whose reported OMA1 staining is high, rather than against background alone (HPA: High in kidney tubule cells; general IHC practice).
How should I score OMA1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then record the percentage of positive cells and intensity to calculate an H-score consistently across sections (general IHC practice). Score kidney tubule cells and colon glandular cells separately if those tissues are used, since their reported staining levels differ (HPA: High in kidney tubule cells; Medium in colon glandular cells). Normalise counts to the number of eligible intact cells, or report positive-cell density per mm² of viable tissue when cell counts cannot be resolved reliably (general IHC practice). Keep retrieval, DAB development, imaging, and threshold settings comparable across slides, and exclude folds, necrosis, and section edges before analysis (general IHC practice).
How can I distinguish genuine OMA1 signal from tissue artefact?
Look for reproducible cell-associated staining in the expected cytoplasmic region, with kidney tubule cells providing a reported high-staining reference (HPA: cytoplasmic tissue profile; High in kidney tubule cells). A DAB deposit confined to section edges, folds, or necrotic regions is suspect, as is signal reproduced in a no-primary control after the same chromogenic steps (general IHC practice). Check endogenous-peroxidase blocking when staining appears widespread, since the selected detection workflow uses a peroxidase-conjugated secondary and DAB (datasheet A06260; general IHC practice). Do not equate staining intensity with OMA1 protease activity: the record describes stress-responsive cleavage of substrates including OPA1 and DELE1, which IHC intensity alone does not measure (UniProt Q96E52 function; general IHC practice).
Boster reagents
Best OMA1 / Metalloendopeptidase OMA1, mitochondrial IHC Antibodies
The catalog antibody has IHC data from human paraffin sections and IF data from human cells; its listed reactivity includes human, mouse, and rat (A06260 catalog; image captions).
Real IHC data
Anti-OMA1 Antibody ®
Cat # A06260
A06260 has IHC images from paraffin sections of human colon and ovarian cancers (A06260 IHC captions). A06260 also has an IF image from A549 cells and lists IHC, IF, and ICC among its applications (A06260 IF caption; catalog applications).
Which to pick: For tissue IHC, choose A06260: its IHC captions document paraffin sections, EDTA retrieval at pH 8.0, and primary antibody at 2 μg/ml (A06260 IHC captions); the fixative is unreported (A06260 IHC captions). For IF/ICC, the same SKU has an A549 cell IF image using 5 μg/ml primary antibody (A06260 IF caption). For mouse or rat samples, A06260 lists reactivity with both species, but the supplied IHC and IF images show human samples; clonality is unreported (A06260 catalog reactivity; image captions; catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96E52 (OMA1_HUMAN, Metalloendopeptidase OMA1, mitochondrial).
- Human Protein Atlas. OMA1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. OMA1 subcellular location (ICC-IF): Localized to the nucleoplasm and mitochondria..
- Human Protein Atlas. OMA1 antibody validation summary (1 antibodies).
- TLR4 Activation Promotes the Progression of Experimental Autoimmune Myocarditis to Dilated Cardiomyopathy by Inducing Mitochondrial Dynamic Imbalance. Oxidative medicine and cellular longevity 2018 — PMC6038665.
- Long Noncoding RNA PCGEM1 Facilitates Tumor Growth and Metastasis of Osteosarcoma by Sponging miR-433-3p and Targeting OMA1. Orthopaedic surgery 2023 — PMC10102293.
- Inhibition of mitochondrial OMA1 ameliorates osteosarcoma tumorigenesis. Cell death & disease 2024 — PMC11530700.
- OMA1 competitively binds to HSPA9 to promote mitophagy and activate the cGAS-STING pathway to mediate GBM immune escape. Journal for immunotherapy of cancer 2024 — PMC11015223.
- PubMed PMID:12886954 — UniProt-cited evidence.
- PubMed PMID:16710414 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for OMA1 (Metalloendopeptidase OMA1, mitochondrial). Expected localisation Cytoplasmic staining (HPA tissue IHC); mitochondrial inner membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06260). Validated IHC antibody: A06260 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet A06260); 2. Fixation: Image fixative and duration unreported (datasheet A06260); 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 A06260); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06260); 8. Primary antibody: Rabbit anti-OMA1, 2-5 μg/ml (datasheet A06260); 9. Primary incubation: Overnight at 4 °C (datasheet A06260). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Skeletal myocytes undetected despite reported strong expression (HPA tissue IHC; UniProt); Isoform / epitope: 2 isoforms; precursor processing may affect epitopes (UniProt); Regulation: Stress activates OMA1; staining effect unknown (UniProt). Evidence: UniProt Q96E52, HPA tissue IHC · Approved, HPA ICC-IF · nucleoplasm and mitochondria., Datasheet A06260, PMC6038665, PMC10102293. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/OMA1/immunohistochemistry-ihc-guide.