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NR5A2 / Nuclear receptor subfamily 5 group A member 2 · IHC design guide
Design Immunohistochemistry for NR5A2
Plan paraffin-section NR5A2 IHC around widespread nuclear staining (HPA tissue IHC) and the catalog antibody’s 2–5 μg/ml IHC range (datasheet A01332-1). Compare staining with appropriate controls while accounting for its low consistency with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NR5A2 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 staining in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01332-1) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Low consistency between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Abundant in pancreas; lower in liver (UniProt) | |
| Isoform / epitope | 4 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended NR5A2 IHC & IF Protocols
The catalog antibody protocol uses EDTA pH 8.0 heat retrieval (datasheet A01332-1). These 3 articles provide published NR5A2 IHC workflows (PMC6489909; PMC4696807; PMC3573348).
| Sample | Paraffin-embedded human parotid acinar cell carcinoma tissue; fixative not specified (datasheet A01332-1) |
| Fixation | Image fixative and duration unreported (datasheet A01332-1); 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 A01332-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01332-1) |
| Primary antibody | Rabbit anti-NR5A2, 2-5 μg/ml (datasheet A01332-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01332-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A01332-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NR5A2-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Ubiquitous nuclear expression. No signal in the no-primary control. |
💡Decision noteFor paraffin sections, start with EDTA pH 8.0 heat retrieval (datasheet A01332-1). If optimization is needed, compare the published Tris-EDTA or formamide conditions (PMC3573348).
Section 2
What Is the Expected NR5A2 Staining Pattern?
NR5A2 is a nuclear and chromosome-associated protein with no transmembrane segment (UniProt O00482: location and topology). In paraffin-section IHC, expect nuclear staining in cells documented by HPA, including adipocytes, glandular cells and hematopoietic cells (HPA: High in these cell types). HPA describes ubiquitous nuclear expression, but rates its tissue IHC evidence Approved with low consistency between antibody staining and RNA expression (HPA: tissue IHC reliability and profile).
| Distinct chromogenic signal overlaps nuclei in HPA-documented cells, while surrounding tissue remains relatively clear. | This fits the expected compartment (UniProt O00482: nucleus; HPA: ubiquitous nuclear expression). HPA reports High staining in adipocytes and in glandular cells from several tissues (HPA: tissue IHC). Judge the signal against nuclear morphology and controls; HPA's Approved rating carries a low staining–RNA consistency caveat (HPA: reliability description). |
| Strong staining is predominantly cytoplasmic, membranous or extracellular, with little nuclear signal. | Treat this as a possible staining artefact or off-target signal because NR5A2 is nuclear/chromosome-associated and has no transmembrane segment (UniProt O00482: location and topology). Check whether the apparent signal follows tissue pigment, section edges or detection background using routine IHC controls (standard IHC practice). |
| Chromogen appears mainly in a cell population outside the HPA-documented examples, especially without convincing nuclear staining. | Consider antibody cross-reactivity or endogenous detection activity (standard IHC practice). Cell identity alone cannot establish an error: HPA describes ubiquitous nuclear expression and supplies no negative cell-type list (HPA: tissue IHC profile). Compare localisation and appropriate detection controls before calling the population NR5A2-positive (UniProt O00482: nucleus; standard IHC practice). |
| Weak, diffuse colour covers many compartments or obscures nuclear boundaries. | Diffuse background does not establish NR5A2 expression; the expected readout is nuclear (UniProt O00482: location; HPA: nuclear IHC profile). Review background controls, blocking, antibody concentration and chromogen development as general IHC variables (standard IHC practice). HPA's High tissue calls should not be used to excuse indiscriminate colour (HPA: tissue IHC levels). |
| No nuclear signal appears in an HPA-reported High cell population, such as adipocytes in adipose tissue. | The run may lack a detectable positive signal, but one negative section does not establish biological absence (HPA: High in adipocytes; HPA: low staining–RNA consistency). First assess tissue preservation and the assay's positive and detection controls (standard IHC practice). Interpret any remaining negative result in light of HPA's reliability caveat (HPA: tissue IHC reliability). |
💡Expected NR5A2 appearanceCall a positive result when clear nuclear chromogen is present in identifiable HPA-documented cells, with potentially High staining in listed populations; cytoplasmic-only colour or diffuse colour lacking nuclear boundaries is suspect (UniProt O00482: nucleus; HPA: High tissue IHC calls; standard IHC practice).
| Subcellular location | Score nuclear localisation as the principal IHC pattern (UniProt O00482: nucleus and chromosome; HPA: ubiquitous nuclear expression). No transmembrane segment or signal peptide is listed, so a membrane or secreted pattern has no support here (UniProt O00482: topology and processing). |
| Tissue and cell context | HPA reports High IHC staining in adipocytes, adrenal/appendix/breast glandular cells, marrow hematopoietic cells, bronchial respiratory epithelium, caudate glia and cerebellar granular-layer cells (HPA: tissue IHC). These examples guide slide review; the supplied HPA extract lists no negative tissues or cells (HPA: tissue IHC). |
| Confidence in the observed pattern | The HPA tissue IHC assessment is Approved, while antibody staining has low consistency with RNA expression (HPA: reliability description). The listed antibodies HPA005455 and HPA017067 are IHC Approved; neither is marked Enhanced in the supplied record (HPA: antibody validation). Avoid treating any single stained population as independently confirmed by RNA (HPA: reliability description). |
| Isoforms and epitope coverage | UniProt lists four NR5A2 isoforms (UniProt O00482: isoforms 1–4). The supplied record gives no antibody epitope or isoform-specific IHC validation, so it cannot establish which isoforms contribute to a stain (UniProt O00482: isoforms; HPA: supplied antibody validation). |
| IF/ICC Q&A: should puncta be expected? | HPA's ICC-IF summary places NR5A2 in nuclear speckles but labels the main location uncertain (HPA: subcellular ICC-IF). For this paraffin-section IHC guide, assess nuclear staining; do not require resolvable speckles for a positive chromogenic result (UniProt O00482: nucleus; HPA: tissue IHC profile). |
| Situation | Likely cause | Next action |
|---|---|---|
| Nuclear signal is absent from HPA-reported High adipocytes or glandular cells. | A weak or failed IHC run is possible; HPA's staining–RNA consistency caveat also limits certainty for a single specimen (HPA: High tissue calls and reliability description). | Check a documented positive section, detection reagents and counterstained nuclear morphology; review antigen-retrieval and antibody-incubation settings as general workflow variables (standard IHC practice). Do not infer NR5A2-specific retrieval sensitivity from these sources (HPA: tissue IHC; UniProt O00482). |
| Most colour lies outside nuclei. | Non-nuclear staining conflicts with the reported NR5A2 location and may reflect nonspecific staining or detection background (UniProt O00482: nucleus; standard IHC practice). | Compare with a no-primary control and a documented positive section; assess where chromogen falls relative to nuclei (standard IHC practice; HPA: nuclear IHC profile). |
| Unexpected cells stain strongly. | Cross-reactivity or endogenous detection activity is possible, but HPA reports ubiquitous nuclear expression and provides no negative cell-type list (standard IHC practice; HPA: tissue IHC profile). | Confirm cell identity and nuclear localisation; use an appropriate detection control before assigning NR5A2 expression (standard IHC practice; UniProt O00482: nucleus). |
| Diffuse background makes individual nuclei hard to score. | Blocking, antibody concentration or chromogen development may contribute to background in chromogenic IHC (standard IHC practice). | Check background controls and optimise those general assay variables so nuclear boundaries can be judged (standard IHC practice; HPA: nuclear IHC profile). |
| Two slides show different proportions of positive cells. | HPA's Approved tissue IHC assessment includes low consistency between antibody staining and RNA expression; the extract does not provide a universal positive-cell threshold (HPA: reliability and tissue IHC). | Compare equivalent cell populations, section quality and control performance before interpreting a biological difference (standard IHC practice). Report the observed nuclear distribution rather than an unsupported cutoff (UniProt O00482: nucleus; HPA: tissue IHC). |
| ICC-IF shows punctate nuclei while chromogenic IHC appears uniformly nuclear. | HPA calls nuclear speckles in ICC-IF uncertain, whereas its tissue IHC profile is broadly nuclear (HPA: subcellular ICC-IF and tissue IHC). | Assess each image by its own stated evidence: nuclear localisation supports IHC interpretation, and speckle resolution is not required for chromogenic scoring (UniProt O00482: nucleus; HPA: tissue IHC profile and ICC-IF uncertainty). |
Experiment evidence
IHC & IF images showing NR5A2 staining
A01332-1 · IHCIHC analysis of NR5A2 using anti-NR5A2 antibody (A01332-1). NR5A2 was detected in a paraffin-embedded section of human parotid acinar cell c
A01332-1 · IHCIHC analysis of NR5A2 using anti-NR5A2 antibody (A01332-1). NR5A2 was detected in a paraffin-embedded section of human bladder urothelial ca
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000116833-NR5A2/tissue/cerebral+cortex#imid_2192728
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000116833-NR5A2/tissue/colon#imid_2192638
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000116833-NR5A2/tissue/liver#imid_2192655
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000116833-NR5A2/tissue/kidney#imid_2192640
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005455; green = NR5A2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA005455; green = NR5A2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NR5A2 IHC & IF
🧪Run breast first and assess nuclear staining in glandular cells (HPA: High in breast glandular cells; UniProt O00482: nucleus). HPA detects NR5A2 in all 45 scored tissues, so use no-primary and isotype controls in place of a negative tissue; cells used as internal background checks should lack convincing nuclear DAB signal, but their NR5A2-negative status requires independent validation (HPA: no negative tissue rows; UniProt O00482: nucleus).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: NR5A2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NR5A2 in A-431, A-549, U2OS, with annotated localisation: Nuclear speckles (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only section, an isotype-matched rabbit IgG control, and a matched NR5A2 knockout specimen as a biological negative (selected A01332-1 caption: rabbit primary and peroxidase-conjugated anti-rabbit secondary). Block endogenous peroxidase and inspect any residual DAB signal in the breast section against the no-primary control (selected A01332-1 caption: peroxidase detection with DAB).
⚠️Feasibility: The selected paraffin-section example uses heat retrieval in EDTA at pH 8.0, followed by 2 μg/ml primary antibody overnight at 4°C; its fixative is unreported (selected A01332-1 tissue-IHC caption). No target-specific fixation window or fixation effect is reported in the supplied evidence, and the paraffin-section caption does not establish fixation (selected A01332-1 tissue-IHC caption). The evidence does not establish whether frozen sections or IF are easier; for breast IHC, distinguish nuclear glandular-cell staining from endogenous peroxidase background using the no-primary section (HPA: High in breast glandular cells; UniProt O00482: nucleus; selected A01332-1 caption: DAB detection). The selected A01332-1 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A01332-1).
HPA tissue IHC evidence for NR5A2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: NR5A2 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced NR5A2 IHC Tips
Troubleshoot NR5A2 staining in paraffin sections by checking retrieval, nuclear localisation, controls, and scoring before interpreting chromogenic signal.
How should I retrieve NR5A2 antigen when nuclear staining is weak?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01332-1). The selected antibody's tissue image used this retrieval before overnight incubation at 4°C with 2 μg/ml primary antibody (caption A01332-1). If nuclear signal is weak, compare adjacent sections across recorded heating times while keeping antibody concentration and DAB development constant (standard IHC practice). Assess preservation of tissue structure alongside nuclear signal, because excessive heating can damage sections and make apparent gains difficult to interpret (standard IHC practice).
Could fixation explain inconsistent NR5A2 staining between paraffin blocks?
The selected tissue-IHC caption does not state a fixative, so target-specific fixation sensitivity for this antibody is unknown (caption A01332-1). Record each block's fixative, time before fixation, fixation duration, and processing history before attributing differences to NR5A2 abundance (standard IHC practice). Compare adjacent sections using the documented EDTA retrieval at pH 8.0 and the same 2 μg/ml primary concentration, with a shared reference block in each run (caption A01332-1; standard IHC practice). If staining still differs, report the processing differences and verify tissue integrity before interpreting a biological change (standard IHC practice).
Which staining compartment should count as NR5A2 positive?
Score convincing nuclear staining in intact cells: NR5A2 is annotated in the nucleus and on chromosomes, with no transmembrane segment (UniProt O00482). HPA describes ubiquitous nuclear tissue staining and a possible nuclear-speckle pattern, but marks the speckle assignment uncertain (HPA tissue IHC; HPA subcellular). Examine the hematoxylin counterstain to distinguish nuclear DAB from pigment overlying nuclei, and score compartments separately if both appear (standard IHC practice). Predominantly cytoplasmic or membrane staining warrants review of the negative controls and detection conditions before it is attributed to NR5A2 (UniProt O00482; standard IHC practice).
Can this stain distinguish NR5A2 isoforms or reveal a masked epitope?
NR5A2 has 4 listed isoforms, and its nuclear receptor ligand-binding domain spans residues 300–539 of the 541-residue canonical sequence (UniProt O00482). The supplied caption does not identify the antibody's epitope or establish which isoforms it detects, so nuclear DAB cannot assign an isoform (caption A01332-1). If an expected region stains weakly, first compare retrieval and section quality against a concurrently processed reference section (datasheet A01332-1; standard IHC practice). Any proposed isoform-specific interpretation needs independently documented epitope coverage or an orthogonal isoform assay; staining intensity alone cannot establish it (standard IHC practice).
How should I investigate NR5A2 localisation by multiplex IF?
For a separate IF investigation, pair NR5A2 with an acinar-cell marker when examining pancreas, where NR5A2 is reported as abundant (UniProt O00482). Select spectrally separated fluorophores, consider a far-red channel where tissue autofluorescence is troublesome, and image unstained and single-label controls before interpreting overlap (standard IF practice). Because NR5A2 is nuclear and lacks a transmembrane segment, use controlled permeabilisation to give antibody access to the nuclear epitope, then check that nuclear morphology remains intact (UniProt O00482; standard IF practice). The paraffin IHC caption provides no IF fixation or permeabilisation settings, so establish those conditions within the IF experiment (caption A01332-1).
What should I check when DAB obscures nuclear NR5A2 staining?
The selected paraffin-section image used 10% goat-serum blocking, 2 μg/ml primary antibody overnight at 4°C, and an HRP-based DAB readout (caption A01332-1). Include a primary-omission control and check for endogenous peroxidase activity before assigning diffuse brown signal to NR5A2 (standard IHC practice). If background persists, titrate primary concentration around the documented setting, review blocking and washes, and hold DAB development time consistent across comparison sections (caption A01332-1; standard IHC practice). Evaluate nuclear contrast against the counterstain rather than treating increased overall brown intensity as improved target detection (UniProt O00482; standard IHC practice).
How can I compare NR5A2 IHC across samples reliably? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and score nuclear staining only, consistent with NR5A2's annotated localisation (UniProt O00482). Report the percentage of evaluable nuclei at each intensity from 0–3, or calculate a nuclear H-score from 0–300 by summing intensity times percentage (standard IHC practice). Normalize counts to evaluable nuclei within the same annotated tissue compartment, and exclude damaged edges and necrotic regions using a rule set before scoring (standard IHC practice). Keep retrieval, antibody concentration, DAB development, and image thresholds consistent; HPA reports low agreement between antibody staining and RNA data, so avoid treating a small score difference as definitive biology (caption A01332-1; HPA tissue IHC; standard IHC practice).
How do I separate true NR5A2 signal from artefact?
A plausible positive has staining within intact nuclei, consistent with NR5A2's nuclear and chromosome localisation (UniProt O00482). Compare the stained cell type with the tissue context: UniProt reports abundant expression in pancreas and lower expression in liver, while HPA describes widespread nuclear staining with low agreement between staining and RNA data (UniProt O00482; HPA tissue IHC). Treat staining concentrated at cut edges, in necrotic areas, or outside nuclei as a reason to inspect morphology and controls before scoring (standard IHC practice). A primary-omission control and endogenous-peroxidase check help identify detection-related DAB signal; replicate nuclear staining in comparable intact regions strengthens the interpretation (standard IHC practice).
Boster reagents
Best NR5A2 / Nuclear receptor subfamily 5 group A member 2 IHC Antibodies
One human-reactive anti-NR5A2 antibody has real chromogenic IHC data from paraffin-embedded human carcinoma sections; no IF/ICC data are supplied (catalog applications and image captions).
Real IHC data
Anti-NR5A2 Antibody ®
Cat # A01332-1
A01332-1 will render with its own IHC figure from a paraffin-embedded human parotid acinar cell carcinoma section (catalog figure caption). Additional catalog captions show IHC staining in paraffin-embedded human bladder urothelial carcinoma and colorectal adenocarcinoma sections (catalog image captions).
Which to pick: For human tissue IHC, choose A01332-1: its paraffin-section caption reports EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody; the fixative is unreported (catalog figure caption). No IF/ICC or cross-species option is documented, because A01332-1 lists IHC and human reactivity only for those choices (catalog applications and reactivity). A01332-1 is a rabbit antibody, and its clonality is unreported (catalog host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00482 (NR5A2_HUMAN, Nuclear receptor subfamily 5 group A member 2).
- Human Protein Atlas. NR5A2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NR5A2 subcellular location (ICC-IF): Localized to the nuclear speckles..
- Human Protein Atlas. NR5A2 antibody validation summary (2 antibodies).
- Nr5a2 promotes tumor growth and metastasis of gastric cancer AGS cells by Wnt/beta-catenin signaling. OncoTargets and therapy 2019 — PMC6489909.
- Dysfunction of Liver Receptor Homolog-1 in Decidua: Possible Relevance to the Pathogenesis of Preeclampsia. PloS one 2015 — PMC4696807.
- NR5A2 gene affects the overall survival of LUAD patients by regulating the activity of CSCs through SNP pathway by OCLR algorithm and immune score. Heliyon 2024 — PMC11004709.
- Comparison of the transcriptomes of long-term label retaining-cells and control cells microdissected from mammary epithelium: an initial study to characterize potential stem/progenitor cells. Frontiers in oncology 2013 — PMC3573348.
- PubMed PMID:9786908 — UniProt-cited evidence.
- PubMed PMID:10359768 — UniProt-cited evidence.
- PubMed PMID:11595170 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NR5A2 (Nuclear receptor subfamily 5 group A member 2). Expected localisation Nuclear staining in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01332-1). Validated IHC antibody: A01332-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human parotid acinar cell carcinoma tissue; fixative not specified (datasheet A01332-1); 2. Fixation: Image fixative and duration unreported (datasheet A01332-1); 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 A01332-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01332-1); 8. Primary antibody: Rabbit anti-NR5A2, 2-5 μg/ml (datasheet A01332-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01332-1). Watch for: Fixation: Keep fixation conditions consistent across sections (standard IHC practice; not target-specific); Caveat: Low consistency between staining and RNA expression (HPA tissue IHC); Isoform / epitope: 4 isoforms; epitope coverage is unspecified (UniProt); Regulation: Abundant in pancreas; lower in liver (UniProt). Evidence: UniProt O00482, HPA tissue IHC · Approved, HPA ICC-IF · nuclear speckles., Datasheet A01332-1, PMC6489909, PMC4696807. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NR5A2/immunohistochemistry-ihc-guide.