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NRF1 / Nuclear respiratory factor 1 · IHC design guide
Design Immunohistochemistry for NRF1
Plan paraffin-section NRF1 IHC around the nuclear staining reported in most tissues (HPA tissue IHC). Use the tissue profile to select comparison sections and interpret the skeletal muscle discrepancy (HPA tissue IHC; UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NRF1 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 | Nuclear in most tissues (HPA tissue IHC) | |
| Staining pattern | Nuclear staining across most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01129-2) | |
| Positive control | Adrenal gland+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 compared paraffin sections. (selected-SKU IHC image A01129-2) | |
| Caveat | Skeletal myocytes were not detected despite reported strong muscle expression (HPA tissue IHC; UniProt) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | Four isoforms; check whether the epitope is shared (UniProt) |
Section 1
Recommended NRF1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with a published NRF1 protocol for breast cancer tissue microarrays (PMC5774493).
| Sample | Paraffin-embedded human ovary cancer tissue; fixative not specified (datasheet A01129-2) |
| Fixation | Image fixative and duration unreported (datasheet A01129-2); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01129-2) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01129-2) |
| Primary antibody | Rabbit anti-NRF1, 0.5-1μg/ml (datasheet A01129-2) |
| Primary incubation | Overnight at 4 °C (datasheet A01129-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01129-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NRF1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A01129-2); the published breast cancer protocol also used microwave citrate retrieval at pH 6.0 (PMC5774493).
Section 2
What Is the Expected NRF1 Staining Pattern?
NRF1 is a nuclear transcription factor with no transmembrane segment (UniProt Q16656: nucleus, topology). In paraffin-section IHC, expect predominantly nuclear staining across many tissues, including glandular cells in colon, duodenum and breast and glial cells in cerebral cortex (HPA: tissue IHC). HPA rates the tissue staining Approved, with medium consistency against RNA data and external verification pending (HPA: tissue IHC reliability).
| Predominantly nuclear staining of colon or duodenum glandular cells, or cerebral cortex glial cells, at a medium level (HPA: tissue IHC). | This matches the observed cell types and intensity in those tissues (HPA: tissue IHC) and the expected nuclear compartment (UniProt Q16656: nucleus). Compare nuclei with adjacent cells on the same section; the tissue pattern is Approved but awaits external verification (HPA: tissue IHC reliability). |
| Strong staining is confined to cytoplasm, with little or no nuclear signal (HPA: nucleoplasm is the supported ICC-IF location). | Treat a cytoplasm-dominant IHC result as a localisation mismatch and investigate possible artefact (UniProt Q16656: nucleus; HPA: subcellular ICC-IF). HPA also reports cytosol in ICC-IF, but labels that additional location uncertain; it does not establish cytoplasm-only paraffin IHC as the expected pattern (HPA: subcellular ICC-IF). |
| Prominent staining appears in skeletal-muscle myocytes or adipocytes (HPA: both cell types not detected by tissue IHC). | Check cell identity and controls before assigning the signal to NRF1; cross-reactivity or endogenous detection activity is possible (general IHC practice). UniProt reports strongest expression in skeletal muscle at the tissue level, while HPA reports myocytes not detected by IHC, so this discrepancy alone cannot validate or disprove a stain (UniProt Q16656: tissue specificity; HPA: skeletal-muscle IHC). |
| Diffuse color covers nuclei and surrounding tissue without a clear cellular boundary (general IHC practice). | An indistinct deposit is difficult to score as NRF1-positive nuclear staining (UniProt Q16656: nucleus; general IHC practice). Assess background with an appropriate negative control and review blocking, washes and chromogenic detection conditions (general IHC practice); HPA's tissue profile does not identify the cause of diffuse background (HPA: tissue IHC). |
| No nuclear signal appears in colon glandular cells despite intact tissue morphology (HPA: colon glandular cells medium by IHC). | A missing signal in this HPA-observed cell population warrants a run-level check before calling the specimen NRF1-negative (HPA: colon tissue IHC; general IHC practice). The HPA rating is Approved with medium RNA–staining consistency, so one negative result cannot by itself establish absence of NRF1 (HPA: tissue IHC reliability). |
💡Expected NRF1 appearanceCall the result consistent with NRF1 when colon or duodenum glandular cells show predominantly nuclear, medium-level staining (HPA: tissue IHC; UniProt Q16656: nucleus); strong cytoplasm-only staining or prominent adipocyte staining is a pattern to investigate (HPA: subcellular ICC-IF; HPA: adipocytes not detected by IHC).
| Tissue and cell choice (HPA: tissue IHC) | HPA reports medium staining in several glandular populations and brain glial cells, low staining in lung macrophages, and no detected staining in adipocytes or skeletal-muscle myocytes (HPA: tissue IHC). Choose comparison areas by cell type, since a tissue-wide label can hide differences among its cells (general IHC practice). |
| Antibody evidence (HPA: HPA029329 IHC Approved) | HPA lists HPA029329 as IHC Approved and the tissue profile as Approved, with medium agreement between staining and RNA expression and external verification pending (HPA: antibody validation; HPA: tissue IHC reliability). Interpret a discordant slide with that evidence limit in mind; no IHC-Enhanced status is supplied (HPA: antibody validation). |
| Isoforms and modifications (UniProt Q16656) | UniProt lists four NRF1 isoforms and five example CK2 phosphoserine sites, but the supplied record gives no antibody epitope or isoform coverage (UniProt Q16656: isoforms, modified residues). These annotations alone cannot predict which isoforms a stain detects or whether modification changes IHC signal (UniProt Q16656; general IHC interpretation). |
| Does IF/ICC show the same compartment? (HPA: subcellular ICC-IF) | In ICC-IF, HPA supports nucleoplasm as the main location and labels additional cytosol localisation uncertain; images are listed for A-431 and U-251MG (HPA: subcellular ICC-IF). This supports a nuclear localisation check, while the paraffin-section tissue and intensity expectations come from HPA tissue IHC (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known HPA-positive cells lack nuclear color (HPA: colon glandular cells medium by IHC). | A run or detection failure is possible; a single blank section cannot identify its cause (general IHC practice). | Confirm the positive-control section, then review antigen retrieval, antibody dilution, detection reagents and counterstain visibility (general IHC practice). No NRF1-specific retrieval condition or dilution is supplied (HPA: supplied IHC record). |
| Cytoplasmic signal overwhelms nuclear signal (UniProt Q16656: nucleus). | The stain conflicts with the supported main localisation; HPA's extra cytosol ICC-IF location is uncertain (HPA: subcellular ICC-IF). | Compare nuclear detail and background controls, then reassess staining specificity (general IHC practice). Do not use the uncertain ICC-IF cytosol observation to accept cytoplasm-only IHC (HPA: subcellular ICC-IF). |
| Strong signal appears in adipocytes or skeletal-muscle myocytes (HPA: not detected by IHC). | Cell identification, cross-reactivity or endogenous detection activity may explain unexpected color (general IHC practice). | Verify the stained cell type and inspect appropriate negative and detection controls (general IHC practice). Keep the UniProt skeletal-muscle expression statement distinct from HPA's myocyte IHC observation (UniProt Q16656: tissue specificity; HPA: skeletal-muscle IHC). |
| Diffuse chromogen obscures nuclei (general IHC practice). | Background from blocking, washes or detection chemistry is possible (general IHC practice). | Review negative-control staining and the blocking, wash and chromogen steps before scoring nuclear signal (general IHC practice). HPA's NRF1 profile does not establish a target-specific background mechanism (HPA: tissue IHC). |
| A low-staining tissue gives an equivocal result (HPA: lung macrophages low by IHC). | A weak HPA-observed signal can be difficult to separate from background on a single slide (HPA: tissue IHC; general IHC practice). | Compare the same cell population with a medium-staining tissue area and the run's negative control (HPA: tissue IHC; general IHC practice). Avoid treating an equivocal macrophage result alone as proof of absence (HPA: lung macrophages low by IHC). |
| The IHC pattern disagrees with ICC-IF images (HPA: tissue IHC; HPA: subcellular ICC-IF). | The records describe different preparations, and HPA labels the ICC-IF cytosol location uncertain (HPA: subcellular ICC-IF). | Score paraffin tissue against HPA's observed cell types and nuclear tissue profile; use ICC-IF to inform the supported main compartment (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing NRF1 staining
A01129-2 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (A01129-2). NRF1 was detected in paraffin-embedded section of human ovary cancer tissue. Heat
A01129-2 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (A01129-2). NRF1 was detected in paraffin-embedded section of mouse intestine tissue. Heat med
M01129-1 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (M01129-1). NRF1 was detected in paraffin-embedded section of human rectal cancer tissues. Hea
M01129-1 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (M01129-1). NRF1 was detected in paraffin-embedded section of human lung cancer tissues. Heat
M01129-1 · IFIF analysis of NRF1 using anti-NRF1 antibody (M01129-1). NRF1 was detected in immunocytochemical section of MCF7 cells. Enzyme antigen retri
M01129 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (M01129). NRF1 was detected in a paraffin-embedded section of human bladder cancer tissue. Hea
M01129 · IHCIHC analysis of NRF1 using anti-NRF1 antibody (M01129). NRF1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat
PA1948 · IHCAnti-NRF1 antibody, PA1948, IHC(P) IHC(P): Human Mammary Cancer Tissue
PA1948 · IHCAnti-NRF1 antibody, PA1948, IHC(P) IHC(P): Rat Intestine Tissue
PA1948 · IFAnti-NRF1 antibody, PA1948, ICC ICC: A549 Cell
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000106459-NRF1/tissue/cerebral+cortex#imid_8286708
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000106459-NRF1/tissue/colon#imid_8067198
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000106459-NRF1/tissue/liver#imid_8067191
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000106459-NRF1/tissue/kidney#imid_8067200
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029329; green = NRF1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA029329; green = NRF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NRF1 IHC & IF
🧪Run adrenal gland first: its glandular cells should show nuclear NRF1 staining (HPA: Medium in adrenal gland glandular cells; UniProt Q16656: nucleus). Use adipose tissue as the negative comparator (HPA: Not detected in adipocytes); on the adrenal slide, use cells without discernible nuclear signal to assess background, without assuming every nonglandular cell is NRF1-negative.
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NRF1 in A-431, U-251MG, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG control matched to the primary antibody’s clonality when known (selected-SKU IHC caption: rabbit primary). Use an NRF1 knockout specimen as a biological negative; for the caption’s SABC/DAB detection, check endogenous peroxidase and biotin background on the adrenal section (selected-SKU IHC caption: SABC/DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the exact A01129-2 tissue-IHC caption does not report a fixative. The paraffin-section example used heat retrieval in citrate buffer at pH 6 for 20 minutes; whether retrieval is required across specimens is unreported (selected-SKU IHC caption). Frozen-section or IF ease relative to paraffin IHC is unreported; assess adrenal background with the no-primary control because the caption’s SABC/DAB method can be affected by endogenous peroxidase or biotin (selected-SKU IHC caption: SABC/DAB).
HPA tissue IHC evidence for NRF1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NRF1 IHC Tips
Troubleshoot NRF1 staining in paraffin sections by checking nuclear localisation, retrieval, controls and cell type before comparing signal intensity (UniProt Q16656; HPA tissue IHC).
How should I optimise retrieval when NRF1 nuclear staining is weak?
Start with heat-mediated citrate buffer at pH 6 for 20 minutes in paraffin sections (datasheet A01129-2). If nuclear staining remains weak, compare modest changes in heating time on matched sections while keeping the antibody concentration and detection conditions fixed (standard IHC practice). The product image used 1 µg/mL primary antibody overnight at 4°C, so reproduce those conditions before attributing a weak result to retrieval (datasheet A01129-2). Compare nuclear signal with tissue background and a no-primary control; excessive retrieval can impair morphology and complicate nuclear scoring (standard IHC practice). NRF1 should predominantly localise to nuclei (UniProt Q16656; HPA subcellular).
Could fixation explain variable NRF1 staining between paraffin blocks?
The selected paraffin-section caption does not state a fixative, and target-specific NRF1 fixation sensitivity is unknown from the supplied evidence (datasheet A01129-2). Record each block’s fixative, fixation duration, processing history and section age before comparing staining across cases (standard IHC practice). Run sections from different blocks together using citrate at pH 6 for 20 minutes and a shared detection batch to reduce workflow variation (datasheet A01129-2; standard IHC practice). Assess nuclear preservation and staining in comparable cell populations, and avoid treating a poorly preserved region as evidence of absent NRF1 (UniProt Q16656; standard IHC practice). Neither tissue expression patterns nor NRF1 modifications establish a fixation effect (HPA tissue IHC; UniProt Q16656).
Should cytoplasmic staining count as positive NRF1 in chromogenic IHC?
Prioritise a nuclear pattern: NRF1 is annotated as nuclear, and tissue IHC reports nuclear expression in most tissues (UniProt Q16656; HPA tissue IHC). The subcellular atlas supports nucleoplasmic localisation while listing cytosol as an uncertain additional location, so isolated diffuse cytoplasmic DAB warrants caution (HPA subcellular). Compare the suspected signal with a no-primary control, nuclear counterstain and neighbouring well-preserved cells under the same retrieval and exposure conditions (standard IHC practice). Score nuclear and cytoplasmic staining separately, noting cell type and section region rather than combining both compartments into one positive fraction (standard IHC practice). A missing transmembrane segment does not establish a tissue-specific cytoplasmic pattern (UniProt Q16656).
Can this IHC stain distinguish NRF1 isoforms or phosphorylation states?
NRF1 has 4 annotated isoforms—Long, Short, 3 and 4—but the supplied product caption gives no epitope sequence or isoform coverage (UniProt Q16656; datasheet A01129-2). It therefore cannot establish which isoform contributes to nuclear DAB in a paraffin section (UniProt Q16656; datasheet A01129-2). CK2-associated phosphoserines are annotated at residues 39, 44, 46, 47 and 52, yet the caption provides no phospho-specific validation (UniProt Q16656; datasheet A01129-2). Obtain the antibody’s immunogen or epitope information and validate any isoform or modification claim with an appropriate independent assay before interpreting staining differences (standard IHC practice). Keep retrieval and detection identical across the sections being compared (standard IHC practice).
How can I check NRF1 localisation by multiplex IF?
For a separate IF experiment, pair NRF1 with a marker identifying the expected cell population and a nuclear counterstain, then assess overlap at the single-cell level (standard IF practice). NRF1 is primarily nucleoplasmic, while its additional cytosolic location is uncertain, so score those compartments separately (HPA subcellular). Choose fluorophores in channels with low tissue autofluorescence, and inspect unstained and single-stain controls before interpreting faint cytosolic signal (standard IF practice). Because NRF1 has no transmembrane segment and the expected epitope is intracellular, include permeabilisation that permits antibody access to nuclei; optimise its strength for morphology (UniProt Q16656; standard IF practice). The paraffin IHC caption provides no IF fixation validation (datasheet A01129-2).
What causes widespread DAB or stromal background around NRF1-positive nuclei?
First distinguish nuclear DAB from diffuse stromal pigment using the counterstain, a no-primary control and matched imaging settings (UniProt Q16656; standard IHC practice). The selected workflow uses 10% goat serum, a biotinylated secondary antibody, a streptavidin–biotin complex and DAB, so assess nonspecific secondary binding and endogenous biotin (datasheet A01129-2; standard IHC practice). Include a peroxidase block and inspect a no-primary section for residual enzyme-dependent colour; these are general chromogenic controls (standard IHC practice). If background persists, titrate the primary from the documented 1 µg/mL condition and optimise blocking and washes on matched sections (datasheet A01129-2; standard IHC practice). Exclude folds, edges and damaged tissue from interpretation (standard IHC practice).
How should I quantify NRF1 when cell composition varies across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region of interest before scoring, because NRF1 staining is reported across many tissues and cell types (HPA tissue IHC; standard IHC practice). Report nuclear percentage positive and intensity, or a nuclear H-score from 0–300, using the same threshold and counterstain assessment for every slide (standard IHC practice). Normalise positive nuclei to all evaluable nuclei of the specified cell population, or report positive-cell density per mm² of viable analysed tissue (standard IHC practice). Keep retrieval, DAB development and image acquisition consistent, and exclude necrosis, folds and tissue edges from the denominator (standard IHC practice). Report cytoplasmic signal separately because its NRF1 localisation remains uncertain (HPA subcellular).
When is an apparent NRF1-positive region more likely to be artefact?
A convincing result places DAB in intact nuclei of identifiable cells, consistent with NRF1’s nuclear annotation and the reported tissue-IHC pattern (UniProt Q16656; HPA tissue IHC). Treat signal confined to section edges, folds, necrotic regions or no-primary controls as suspect and inspect the counterstain before scoring (standard IHC practice). Diffuse cytoplasmic colour alone needs independent support because nucleoplasmic localisation is supported while cytosolic localisation is uncertain (HPA subcellular). Compare like cell populations: the atlas reports medium staining in several glandular populations but no detectable staining in skeletal-muscle myocytes, despite a separate transcript-level statement of strongest skeletal-muscle expression (HPA tissue IHC; UniProt Q16656). Residual endogenous peroxidase can also create DAB signal (standard IHC practice).
Boster reagents
Best NRF1 / Nuclear respiratory factor 1 IHC Antibodies
The catalog shows NRF1 staining in paraffin sections from human, mouse and rat tissues, plus IF in human MCF7 cells and ICC in A549 cells (catalog image captions).
Real IHC data
Anti-NRF1 Antibody ®
Cat # A01129-2
Real IHC data
Anti-NRF1 Antibody ® (monoclonal, 2G4)
Cat # M01129-1
Real IHC data
Anti-NRF1 Rabbit Monoclonal Antibody
Cat # M01129
Real IHC data
Anti-Nuclear respiratory factor 1 NRF1 Antibody ®
Cat # PA1948
A01129-2 has IHC images of human ovary cancer, mouse intestine and spleen, and rat heart; M01129-1 has human rectal, lung, mammary and testis cancer IHC images plus MCF7 IF (catalog image captions). M01129 has human bladder, breast, clear cell renal cell carcinoma and colon cancer IHC images; PA1948 has human mammary and intestinal cancer and rat intestine IHC images plus A549 ICC (catalog image captions).
Which to pick: For paraffin tissue IHC, choose rabbit monoclonal M01129 if its documented EDTA retrieval at pH 8.0 and 1:50 dilution suit the assay (M01129 IHC image caption; catalog: clone ADGI-14). For IF/ICC, choose mouse monoclonal M01129-1: its MCF7 IF image uses 2 μg/ml, and IF and ICC are listed applications (M01129-1 IF image caption; catalog: clone 2G4 and applications). For IHC across species, A01129-2 has paraffin images from human, mouse and rat with citrate retrieval at pH 6; the captions do not report the fixative (A01129-2 IHC image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q16656 (NRF1_HUMAN, Nuclear respiratory factor 1).
- Human Protein Atlas. NRF1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NRF1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. NRF1 antibody validation summary (1 antibodies).
- NRF1 and NRF2 mRNA and Protein Expression Decrease Early during Melanoma Carcinogenesis: An Insight into Survival and MicroRNAs. Oxidative medicine and cellular longevity 2019 — PMC6794976.
- NRF1 is upregulated by docosahexaenoic acid to ameliorate MASH through the inhibition of ER stress. Cell death & disease 2026 — PMC12811613.
- Increased expression of mitochondrial transcription factor A and nuclear respiratory factor-1 predicts a poor clinical outcome of breast cancer. Oncology letters 2018 — PMC5774493.
- TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells. Scientific reports 2016 — PMC4827396.
- PubMed PMID:8253388 — UniProt-cited evidence.
- PubMed PMID:8034649 — UniProt-cited evidence.
- PubMed PMID:7629110 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NRF1 (Nuclear respiratory factor 1). Expected localisation Nuclear in most tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01129-2). Validated IHC antibody: A01129-2 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovary cancer tissue; fixative not specified (datasheet A01129-2); 2. Fixation: Image fixative and duration unreported (datasheet A01129-2); 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: Citrate pH 6 , 20 min (datasheet A01129-2); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01129-2); 8. Primary antibody: Rabbit anti-NRF1, 0.5-1μg/ml (datasheet A01129-2); 9. Primary incubation: Overnight at 4 °C (datasheet A01129-2). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A01129-2); Caveat: Skeletal myocytes were not detected despite reported strong muscle expression (HPA tissue IHC; UniProt); Isoform / epitope: Four isoforms; check whether the epitope is shared (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q16656, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A01129-2, PMC5774493. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NRF1/immunohistochemistry-ihc-guide.