NDUFA13 / NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13 · IHC design guide

Design Immunohistochemistry for NDUFA13

Plan NDUFA13 IHC in paraffin sections using the granular cytoplasmic tissue pattern as a reference (HPA tissue IHC). The guide covers the IHC-validated antibody, staining controls and interpretation alongside its expected mitochondrial location (UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for NDUFA13 (IHC for NDUFA13): expected localisation Granular cytoplasm (HPA tissue IHC); inner mitochondrial membrane expected (UniProt), antibody A05981-2, validated IHC image, and IHC protocol steps — Printable **NDUFA13** IHC protocol sheet — expected localisation Granular cytoplasm (HPA tissue IHC); inner mitochondrial membrane expected (UniProt), antibody A05981-2, controls and protocol steps. Open the full NDUFA13 IHC guide →

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NDUFA13 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); inner mitochondrial membrane expected (UniProt)
	Staining pattern	Ubiquitous granular cytoplasm; high in glandular cells (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A05981-2)
	Positive control ⓘ	Adrenal gland+4 more · see all
	Negative control ⓘ	None in HPA (detected in all 44 tissues); use no-primary + isotype controls

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A05981-2)
	Caveat	Widespread staining limits negative-tissue controls (HPA tissue IHC)
	Regulation	Reduced in Crohn/UC intestinal mucosa (UniProt)
	Isoform / epitope	2 isoforms; epitope coverage is unspecified (UniProt)

Section 1

Recommended NDUFA13 IHC & IF Protocols

The catalog antibody’s IHC protocol uses heat-mediated EDTA pH 8.0 retrieval (datasheet A05981-2). The published heart-section IHC protocol provides additional staining conditions (PMC13299234).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A05981-2)
Fixation	Image fixative and duration unreported (datasheet A05981-2); 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 A05981-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A05981-2)
Primary antibody	Rabbit anti-NDUFA13, 2-5 μg/ml (datasheet A05981-2)
Primary incubation	Overnight at 4 °C (datasheet A05981-2)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A05981-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	NDUFA13-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 A05981-2); the published IHC excerpt does not specify retrieval (PMC13299234).

Section 2

What Is the Expected NDUFA13 Staining Pattern?

NDUFA13 is mainly mitochondrial and has one transmembrane segment at residues 30–51 in the inner membrane (UniProt Q9P0J0 topology/localisation). In paraffin IHC, expect granular cytoplasmic staining across many cell types, including strong staining in several glandular and neuronal populations (HPA: ubiquitous granular cytoplasmic profile; High in listed cells). The tissue IHC profile is Supported, with high consistency between antibody staining and RNA expression (HPA: reliability Supported).

What am I looking at on my slide?
Granular cytoplasmic staining in colon glandular cells.	A convincing positive pattern: colon glandular cells are High by HPA tissue IHC, and the granular cytoplasm matches its overall profile (HPA: colon; tissue profile). Score cellular staining against adjacent background rather than treating every brown deposit as positive (general IHC practice).
Predominantly nuclear staining with little granular cytoplasm.	Question specificity or interpretation: the usual location is mitochondrial (UniProt Q9P0J0 localisation; HPA: mitochondria supported). UniProt allows possible nuclear translocation after IFN/RA treatment, so a documented treated sample needs separate interpretation (UniProt Q9P0J0 localisation).
A sharply restricted cell population stains while expected glandular cells do not.	Investigate cross-reactivity, endogenous detection activity, and tissue identification (general IHC practice). HPA reports broad expression and High glandular staining in several listed tissues; an unlisted cell type alone is not evidence of off-target binding (HPA: tissue profile; listed High cells).
Uniform haze or precipitate obscures cell boundaries.	This cannot establish NDUFA13 localisation. The supported tissue profile is granular cytoplasmic staining (HPA: tissue profile). Review blocking, washes, reagent concentration, and chromogen development as general IHC variables; compare a matched no-primary control (general IHC practice).
No staining in colon glandular cells or another HPA-listed High population.	Treat the run as inconclusive before calling the sample negative: HPA reports High staining in colon glandular cells, among other listed populations (HPA: colon and listed High cells). Check tissue preservation, retrieval, antibody and detection steps with an appropriate positive control (general IHC practice).

💡Expected NDUFA13 appearanceCall a result positive when cellular granular cytoplasmic signal is clear in an HPA-listed High population, such as colon glandular cells (HPA: tissue profile; colon High); diffuse haze or dominant nuclear-only colour warrants review (HPA: tissue profile; UniProt Q9P0J0 localisation).

How each factor affects the staining
Compartment and resolution	NDUFA13 is an inner-membrane protein with one segment at residues 30–51 (UniProt Q9P0J0 topology). Routine chromogenic IHC supports a granular cytoplasmic readout, not visual assignment to the inner membrane itself (HPA: tissue profile; general IHC practice).
Choice of tissue comparator	HPA lists High glandular staining in colon, duodenum, appendix, breast and adrenal gland, and High staining in selected neuronal populations (HPA: listed High cells). Adipocytes are Low, not a validated negative control (HPA: adipose tissue Low; negative list empty).
Conditional nuclear localisation	UniProt says NDUFA13 localises mainly to mitochondria and may enter the nucleus after IFN/RA treatment (UniProt Q9P0J0 localisation). Record treatment context before interpreting nuclear staining; untreated tissue has no supplied target-specific nuclear expectation.
Isoforms and antibody validation	UniProt annotates 2 isoforms but supplies no isoform-specific staining pattern here (UniProt Q9P0J0 isoforms). HPA041213 is Supported for IHC and ICC; that status does not identify which isoform a given stain represents (HPA: HPA041213 validation).
IF/ICC Q&A: what should fluorescence show?	Expect mitochondrial localisation (HPA: ICC-IF mitochondria supported). HPA041213 is ICC Supported, with images listed for A-431, U-251MG, U2OS and sperm (HPA: ICC-IF; HPA041213 validation). Those ICC observations do not prescribe an IF protocol for paraffin sections.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
A listed High tissue shows no cellular staining.	The result conflicts with HPA's High call for that specific cell population (HPA: listed High cells); a missed IHC step or inadequate assay performance is possible (general IHC practice).	Confirm the expected cell population is present, then review retrieval, primary dilution, detection and a working positive control (general IHC practice). Do not infer NDUFA13-specific fixation sensitivity from this failure.
Signal is diffuse rather than granular.	The observed pattern conflicts with HPA's granular cytoplasmic profile (HPA: tissue profile). Excess reagent, incomplete washing or detection background can blur localisation (general IHC practice).	Compare with a no-primary control; review dilution, washing and chromogen development (general IHC practice). Score only clearly cellular signal with interpretable morphology.
Brown signal persists without primary antibody.	Primary-independent colour may arise from endogenous detection activity or pigment (general chromogenic IHC practice); it does not demonstrate NDUFA13.	Inspect the no-primary slide in the same tissue region; apply the detection system's appropriate endogenous-activity control and distinguish pigment from reaction product (general IHC practice).
Only nuclei stain strongly.	This differs from the usual mitochondrial localisation (UniProt Q9P0J0 localisation; HPA: ICC-IF mitochondria supported), although IFN/RA-associated nuclear translocation is possible (UniProt Q9P0J0 localisation).	Check whether IFN/RA treatment is documented; compare granular cytoplasmic signal and controls before assigning the nuclear colour to NDUFA13 (UniProt Q9P0J0 localisation; general IHC practice).
Adipocytes are faint beside stained cells.	Faint adipocyte staining is compatible with HPA's Low call; HPA supplies no negative tissue in this record (HPA: adipose tissue Low; negative list empty).	Use an HPA-listed High cell population to judge whether the run worked; do not set the positivity threshold by assuming adipocytes must be blank (HPA: listed High cells; adipose tissue Low).
An unexpected cell type stains more strongly than nearby expected cells.	Broad expression means unexpected cell staining alone does not prove cross-reactivity (HPA: ubiquitous tissue profile). Endogenous activity or nonspecific staining remains possible (general IHC practice).	Identify both cell populations, compare their morphology and the no-primary control, and seek independent confirmation before calling a new cell-specific pattern (general IHC practice).

Source: UniProt Q9P0J0 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing NDUFA13 staining

IHC analysis of GRIM19/NDUFA13 using anti-GRIM19/NDUFA13 antibody (A05981-2). GRIM19/NDUFA13 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-GRIM19/NDUFA13 Antibody (A05981-2) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

A05981-2 · IHCIHC analysis of GRIM19/NDUFA13 using anti-GRIM19/NDUFA13 antibody (A05981-2). GRIM19/NDUFA13 was detected in a paraffin-embedded section of

IF analysis of GRIM19/NDUFA13 using anti-GRIM19/NDUFA13 antibody (A05981-2). GRIM19/NDUFA13 was detected in an immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-GRIM19/NDUFA13 Antibody (A05981-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A05981-2 · IFIF analysis of GRIM19/NDUFA13 using anti-GRIM19/NDUFA13 antibody (A05981-2). GRIM19/NDUFA13 was detected in an immunocytochemical section of

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000186010-NDUFA13/tissue/cerebral+cortex#imid_10673839

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000186010-NDUFA13/tissue/colon#imid_10673773

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000186010-NDUFA13/tissue/liver#imid_10673782

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000186010-NDUFA13/tissue/kidney#imid_10673767

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041213; green = NDUFA13, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for NDUFA13 IHC & IF

🧪Run appendix first; its glandular cells should stain (HPA: High in appendix glandular cells). HPA detects NDUFA13 in all 44 scored tissues, so there is no validated negative tissue; use no-primary and isotype controls, and treat unstained cells within the positive slide as relative background rather than proven target-negative cells (HPA: no negative rows; detected in all 44 scored tissues).

Positive control tissue: Adrenal gland (Glandular cells, HPA High)

Negative control tissue: None in HPA: NDUFA13 is detected in all 44 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 NDUFA13 in A-431, U-251MG, U2OS, Sperm, with annotated localisation: Mitochondria (supported) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only slide and a rabbit IgG isotype control matched to the catalog antibody; confirm specificity with NDUFA13 knockout material or an immunizing-peptide block if the peptide is available (caption: rabbit primary antibody; standard IHC practice). In appendix, quench endogenous peroxidase and inspect immune-rich areas for DAB background (standard IHC practice).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A05981-2 paraffin-section caption does not state a fixative (caption: fixative unreported). Heat retrieval in EDTA at pH 8.0 was used for the shown IHC result, but its necessity has not been established (caption: heat-mediated EDTA retrieval). HPA supports mitochondrial ICC-IF localization, but the supplied evidence does not establish whether IF or frozen sections are easier than paraffin IHC; appendix peroxidase background should be checked with the controls (HPA: mitochondria, supported; standard IHC practice).

HPA tissue IHC evidence for NDUFA13

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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	Glandular cells	High	Protein (IHC)	HPA →
Breast	Glandular cells	High	Protein (IHC)	HPA →
Caudate	Neuronal cells	High	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
None in HPA: NDUFA13 is detected in all 44 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.

Section 3

Advanced NDUFA13 IHC Tips

Use the catalog antibody’s paraffin-section IHC evidence to optimize retrieval and detection, then judge staining against NDUFA13’s expected mitochondrial distribution.

What retrieval should I try first for NDUFA13 paraffin-section IHC?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 (datasheet A05981-2). The selected paraffin-section image used this condition before staining human lung cancer tissue (datasheet A05981-2). If staining is weak, compare retrieval duration on adjacent sections while holding antibody concentration and detection conditions constant (standard IHC practice). Include an untreated section to judge whether heating improves the granular cytoplasmic signal expected for NDUFA13 (HPA: ubiquitous granular cytoplasmic expression; standard IHC practice). Avoid treating signal gained only at tissue edges or in damaged areas as successful retrieval; evaluate intact cells throughout the section (standard IHC practice).

Could fixation explain weak NDUFA13 staining in my paraffin sections?

The selected IHC caption identifies paraffin-embedded tissue but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A05981-2). Record the fixative, fixation interval and processing history for each specimen before comparing staining across cases (standard IHC practice). Test adjacent sections with the same EDTA pH 8.0 retrieval and 2 μg/ml primary antibody conditions used in the selected image (datasheet A05981-2; standard IHC practice). Use a consistently processed positive-control section to separate specimen processing differences from a failed staining run (standard IHC practice). Interpret any fixation-related loss as an experimental finding for those specimens, not an established NDUFA13-specific effect (standard IHC practice).

Should NDUFA13 stain nuclei or appear as cytoplasmic granules?

Expect predominantly granular cytoplasmic staining in chromogenic IHC, consistent with the tissue profile and supported mitochondrial localisation (HPA: ubiquitous granular cytoplasmic expression; HPA subcellular: mitochondria supported). NDUFA13 mainly localises to the mitochondrial inner membrane, while nuclear translocation may occur after IFN/RA treatment (UniProt Q9P0J0). Assess nuclear staining separately from cytoplasmic staining rather than combining both into one positive score (standard IHC practice). If nuclear staining dominates an untreated specimen, inspect the negative controls and repeat staining before attributing it to translocation (UniProt Q9P0J0; standard IHC practice). Compare intact cells within the same section so tissue edges and damaged regions do not determine the localisation call (standard IHC practice).

How should isoforms and membrane topology affect antibody troubleshooting?

NDUFA13 has 2 annotated isoforms and a transmembrane segment spanning residues 30–51 (UniProt Q9P0J0). The supplied IHC caption does not identify the catalog antibody’s epitope or establish which isoforms it detects (datasheet A05981-2). Check the antibody’s documented immunogen or epitope before assigning a negative section to isoform absence (standard IHC practice). Compare retrieval conditions on adjacent sections because epitope accessibility in processed tissue must be determined empirically for this antibody (standard IHC practice). If an isoform-specific conclusion matters, validate it with an independent assay that distinguishes the isoforms; a shared staining pattern alone cannot make that assignment (standard IHC practice).

How can I check NDUFA13 localisation by multiplex IF?

For the separate IF/ICC assay, pair NDUFA13 with a marker identifying the expected cell population, such as glandular cells in colon tissue (HPA: high in colon glandular cells; standard IF practice). Choose fluorophores and detection channels after checking the specimen’s autofluorescence, and include single-label controls when assessing overlap (standard IF practice). NDUFA13 contains a transmembrane segment at residues 30–51, but the supplied record does not establish which membrane-facing side the antibody recognises (UniProt Q9P0J0; datasheet A05981-2). Set permeabilisation empirically against the documented epitope’s side when that information is available, while preserving mitochondrial structure (standard IF practice). Judge mitochondrial overlap against the supported subcellular location (HPA subcellular: mitochondria supported).

What should I check when DAB staining looks diffuse or widespread?

First compare the diffuse signal with the expected granular cytoplasmic pattern and an adjacent negative-control section (HPA: ubiquitous granular cytoplasmic expression; standard IHC practice). The selected image used 10% goat serum blocking, 2 μg/ml rabbit primary antibody overnight at 4°C, and peroxidase-based DAB detection (datasheet A05981-2). Check the peroxidase block and a no-primary control to identify endogenous enzyme activity or detection-system background (standard IHC practice). If background persists, assess washing and primary-antibody concentration on matched sections without changing retrieval simultaneously (standard IHC practice). Score only intact cellular staining that remains distinct from pigment, necrosis and tissue-edge deposits (standard IHC practice).

How should I quantify NDUFA13 chromogenic IHC across specimens? ⚠ ANSWER MARKED FOR VERIFICATION

Define the analysis compartment as granular cytoplasmic staining in the relevant intact cells before scoring slides (HPA: ubiquitous granular cytoplasmic expression; standard IHC practice). An H-score combines the percentage of cells at each intensity, while percentage-positive scoring records the stained fraction at a predefined threshold (standard IHC practice). For spatial comparisons, report positive-cell density per mm² of evaluable tissue and state how cells and area were selected (standard IHC practice). Normalise measurements to the counted cells or evaluable tissue area, and keep imaging, threshold and stain-development settings consistent across specimens (standard IHC practice). Record nuclear signal separately if it is under investigation (UniProt Q9P0J0; standard IHC practice).

How do I distinguish true NDUFA13 staining from artefact?

A credible IHC result shows cellular granular cytoplasmic staining consistent with the supported mitochondrial location (HPA: ubiquitous granular cytoplasmic expression; HPA subcellular: mitochondria supported). Compare staining in the cell population being studied with its morphology and the relevant tissue context; glandular cells show high expression in several listed tissues (HPA: tissue IHC profile; standard IHC practice). Exclude staining restricted to section edges, necrotic regions or extracellular deposits from the positive score (standard IHC practice). Check a no-primary control and the peroxidase block when DAB signal could reflect endogenous enzyme activity (standard IHC practice). Interpret predominantly nuclear staining cautiously, since UniProt describes nuclear translocation as possible after IFN/RA treatment (UniProt Q9P0J0).

Boster reagents

Best NDUFA13 / NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13 IHC Antibodies

A05981-2 has IHC images from human paraffin sections and IF images from HeLa cells and human lung cancer tissue (catalog image captions); its listed reactivity covers Human, Mouse and Rat (catalog).

Real IHC data

Anti-GRIM19/NDUFA13 Antibody ®

Cat # A05981-2

View product →

A05981-2 has IHC images from human lung cancer, skin squamous cell carcinoma and urothelial carcinoma paraffin sections, plus IF images from HeLa cells and human lung cancer tissue (catalog image captions). M05981-1 lists IHC and ICC/IF applications with Human reactivity, but provides no IHC or IF image captions (catalog).

Which to pick: Choose A05981-2 for tissue IHC because its own images document staining in human paraffin sections; the fixative is unreported (A05981-2 IHC image captions). Choose A05981-2 for IF/ICC when image evidence matters: its own IF captions show HeLa cells and human lung cancer tissue, while M05981-1 is a rabbit monoclonal, clone 19N74, with listed IHC and ICC/IF applications but no image captions (catalog). For cross-species work, A05981-2 lists Human, Mouse and Rat reactivity, versus Human for M05981-1; A05981-2’s IHC images document human samples only (catalog reactivity; A05981-2 IHC image captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q9P0J0 (NDUAD_HUMAN, NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13).
Human Protein Atlas. NDUFA13 tissue IHC expression (reliability: Supported).
Human Protein Atlas. NDUFA13 subcellular location (ICC-IF): Localized to the mitochondria..
Human Protein Atlas. NDUFA13 antibody validation summary (1 antibodies).
Moderate downregulation of NDUFA13 promotes cardiomyocytes proliferation and heart regeneration through the glycolysis/c‑Myc/Ccnd1 axis. International journal of molecular medicine 2026 — PMC13299234.
Mitochondrial DNA Depletion in Respiratory Chain-Deficient Parkinson Disease Neurons. Annals of neurology 2016 — PMC4819690.
PubMed PMID:10924506 — UniProt-cited evidence.
PubMed PMID:10931946 — UniProt-cited evidence.
PubMed PMID:10810093 — UniProt-cited evidence.

Immunohistochemistry protocol sheet for NDUFA13 (NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13). Expected localisation Granular cytoplasm (HPA tissue IHC); inner mitochondrial membrane expected (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05981-2). Validated IHC antibody: A05981-2 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 44 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 A05981-2); 2. Fixation: Image fixative and duration unreported (datasheet A05981-2); 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 A05981-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05981-2); 8. Primary antibody: Rabbit anti-NDUFA13, 2-5 μg/ml (datasheet A05981-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05981-2). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A05981-2); Caveat: Widespread staining limits negative-tissue controls (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope coverage is unspecified (UniProt); Regulation: Reduced in Crohn/UC intestinal mucosa (UniProt). Evidence: UniProt Q9P0J0, HPA tissue IHC · Supported, HPA ICC-IF · mitochondria., Datasheet A05981-2, PMC13299234. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NDUFA13/immunohistochemistry-ihc-guide.