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ESRRG / Estrogen-related receptor gamma · IHC design guide
Design Immunohistochemistry for ESRRG
This guide covers paraffin-section ESRRG staining, controls and interpretation, with stomach glandular and kidney tubule cells as reported positives (HPA tissue IHC). It flags the mainly cytoplasmic tissue pattern and uncertain agreement between staining and RNA data (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ESRRG 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 | Mainly cytoplasmic in tissue IHC; annotated nuclear (HPA tissue IHC; UniProt) | |
| Staining pattern | Mainly cytoplasmic; stomach glands and kidney tubules stain (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A01470-1) | |
| Positive control | Stomach+1 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01470-1) | |
| Caveat | Staining–RNA discordance makes specificity uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 5 isoforms; verify epitope coverage (UniProt) |
Section 1
Recommended ESRRG IHC & IF Protocols
The catalog antibody uses citrate pH 6 retrieval (datasheet A01470-1). The published IHC protocols below cover mouse embryos, gallbladder tissues, and patient spinal motor neuron sections (PMC3033182; PMC12066295; PMC7885352).
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A01470-1) |
| Fixation | Image fixative and duration unreported (datasheet A01470-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A01470-1) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01470-1) |
| Primary antibody | Rabbit anti-ESRRG, 0.5-1μg/ml (datasheet A01470-1) |
| Primary incubation | Overnight at 4 °C (datasheet A01470-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01470-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ESRRG-positive staining in glandular cells of stomach (HPA tissue IHC: Medium). HPA tissue profile: Mainly cytoplasmic expression in few tissues. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6 retrieval for the catalog antibody (datasheet A01470-1); a published ESRRG IHC protocol also used high-pH retrieval (PMC7885352).
Section 2
What Is the Expected ESRRG Staining Pattern?
ESRRG is a nuclear receptor with no transmembrane segment (UniProt P62508: nucleus, topology). In paraffin-section IHC, HPA reports medium staining in stomach glandular cells and kidney tubule cells, but describes its overall tissue pattern as mainly cytoplasmic and its IHC reliability as Uncertain because staining and RNA data have low consistency (HPA tissue IHC). Interpret compartment and cell identity together; the observed cytoplasmic pattern does not by itself establish specific ESRRG staining.
| Stomach glandular cells or kidney tubule cells show medium chromogenic staining, with identifiable nuclear signal. | These cell populations match HPA's reported IHC positives, and nuclear signal fits UniProt localization (HPA tissue IHC; UniProt P62508: nucleus). Treat the result as a candidate positive: HPA rates tissue IHC Uncertain, so morphology and control sections still matter. |
| Signal is exclusively cytoplasmic, with nuclei remaining clear in the cells being assessed. | HPA describes tissue staining as mainly cytoplasmic, while UniProt places ESRRG in the nucleus and HPA ICC-IF places it in the nucleoplasm (HPA tissue IHC; UniProt P62508; HPA subcellular). Record the mismatch; cytoplasmic staining alone cannot settle target specificity. |
| Strong staining appears in an unexpected cell population, including cells HPA reports as Not detected. | Compare the stained cells with section morphology before assigning identity. HPA reports adipocytes and adrenal glandular cells as Not detected in tissue IHC (HPA tissue IHC). Cross-reactivity or endogenous chromogen-producing activity are possible explanations to test (general IHC practice). |
| Brown signal spreads across nuclei, cytoplasm, and surrounding tissue without a clear cell-level pattern. | This distribution is hard to score as ESRRG because the expected biological compartment is nuclear and the HPA tissue result is uncertain (UniProt P62508: nucleus; HPA tissue IHC: Uncertain). Examine a primary-antibody omission control for detection-system background (general IHC practice). |
| No convincing signal appears in stomach glandular cells or kidney tubule cells. | Both are HPA-reported medium IHC populations, so a blank section prompts a check of tissue identity and assay performance (HPA tissue IHC). It does not prove ESRRG absence: HPA calls tissue IHC Uncertain, and its reported positives are observations, not guaranteed controls. |
💡Expected ESRRG appearanceA candidate positive is cell-associated medium staining in stomach glandular or kidney tubule cells, ideally including nuclei; widespread staining of unrelated cells or control sections suggests a false positive (HPA tissue IHC: Medium, Uncertain; UniProt P62508: nucleus; general IHC practice).
| Compartment evidence | UniProt lists the nucleus, and HPA ICC-IF approves nucleoplasmic localization, but HPA tissue IHC reports mainly cytoplasmic staining (UniProt P62508; HPA subcellular; HPA tissue IHC). This disagreement makes a cytoplasmic-only paraffin-section result an interpretation question, not a confirmed localization. |
| Tissue and antibody evidence | HPA reports medium staining in stomach glandular and kidney tubule cells, yet assigns overall tissue IHC Uncertain reliability; antibody HPA044678 is also IHC Uncertain (HPA tissue IHC; HPA antibodies). Keep the reliability qualification beside any proposed positive tissue. |
| Isoforms and epitope coverage | UniProt lists 5 ESRRG isoforms (UniProt P62508: isoforms 1–5). The supplied record gives no antibody epitope, so it cannot establish which isoforms an IHC stain detects; avoid explaining a tissue difference by isoform recognition without separate evidence. |
| Antigen retrieval | Retrieval conditions are a general paraffin-section IHC variable (general IHC practice). No ESRRG-specific retrieval response or fixation sensitivity is supplied; optimize against appropriate control sections without attributing a stronger or weaker ESRRG signal to a documented target-specific effect. |
| IF/ICC Q&A: where should ESRRG appear? | Nucleoplasm is the approved HPA ICC-IF location, with images listed for A-431, SH-SY5Y, and U2OS; HPA044678 is ICC Approved (HPA subcellular; HPA antibodies). These ICC-IF findings inform compartment interpretation but do not establish the paraffin-section IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The putative positive stomach or kidney section is blank. | A section, retrieval, antibody, or detection problem is possible (general IHC practice); HPA's medium observations are not guaranteed positive controls because tissue IHC is Uncertain (HPA tissue IHC). | Confirm the expected glandular or tubular cells are present, then review reagent controls and the documented IHC conditions before interpreting the blank result (general IHC practice). |
| Only cytoplasm stains in the proposed positive cells. | HPA reports mainly cytoplasmic tissue staining, which conflicts with nuclear UniProt and approved nucleoplasmic ICC-IF localization (HPA tissue IHC; UniProt P62508; HPA subcellular). | Score nuclear and cytoplasmic compartments separately, retain the discordance in the report, and seek independent support before calling cytoplasmic signal specific (general IHC practice). |
| Cells outside the expected population stain strongly. | Cross-reactivity or endogenous detection activity may produce signal in an unexpected population (general IHC practice); HPA reports some populations as Not detected (HPA tissue IHC). | Recheck cell identity on the counterstained section and compare primary-antibody omission and detection controls; evaluate endogenous activity according to the detection chemistry used (general IHC practice). |
| Diffuse chromogen obscures cell boundaries. | Nonspecific antibody binding or detection background can prevent compartment-level scoring (general IHC practice). | Compare the primary-antibody omission control, review blocking and wash conditions, and adjust antibody or detection exposure using the assay documentation (general IHC practice); do not score diffuse color as ESRRG. |
| Staining differs between adjacent areas of the same section. | Section handling or uneven reagent coverage can create a spatial pattern unrelated to cell identity (general IHC practice). | Inspect whether the pattern follows tissue architecture or a section edge, then repeat with uniform reagent coverage if needed (general IHC practice); compare the same cell type across the section. |
| Nuclear staining is difficult to distinguish from the counterstain. | Overlapping chromogen and counterstain can obscure subcellular assignment (general IHC practice), which matters because ESRRG is nuclear in UniProt (UniProt P62508: nucleus). | Review the slide at higher magnification and adjust counterstain intensity in a repeat section if needed (general IHC practice); report an unresolved compartment as indeterminate. |
Experiment evidence
IHC & IF images showing ESRRG staining
A01470-1 · IHCIHC analysis of ESRRG using anti-ESRRG antibody (A01470-1). ESRRG was detected in paraffin-embedded section of mouse brain tissue . Heat med
A01470-1 · IHCIHC analysis of ESRRG using anti-ESRRG antibody (A01470-1). ESRRG was detected in paraffin-embedded section of rat cardiac muscle tissue. He
A01470-1 · IFIF analysis of ESRRG using anti-ESRRG antibody (A01470-1). ESRRG was detected in immunocytochemical section of A549 cell. Enzyme antigen ret
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196482-ESRRG/tissue/cerebral+cortex#imid_12098543
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196482-ESRRG/tissue/colon#imid_12095149
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196482-ESRRG/tissue/liver#imid_12095158
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000196482-ESRRG/tissue/kidney#imid_12095151
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044678; green = ESRRG, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA044678; green = ESRRG, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ESRRG IHC & IF
🧪Run kidney first: tubular cells should show nuclear ESRRG staining (HPA: Medium in kidney tubular cells; UniProt P62508: nucleus). Run adipose tissue as the negative, assessing adipocytes (HPA: Not detected in adipocytes); on the kidney slide, any neighboring cells without nuclear DAB signal should retain counterstain alone, without assuming a specific cell type is ESRRG-negative (standard IHC interpretation).
Positive control tissue: Stomach (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ESRRG in A-431, SH-SY5Y, U2OS, with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit IgG isotype control matched to the primary antibody’s clonality, and ESRRG knockout tissue as a biological negative (A01470-1 caption: rabbit primary; standard IHC controls). For kidney, quench endogenous peroxidase and check for endogenous biotin background when using the caption’s SABC/DAB detection (A01470-1 caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: An ESRRG-specific fixation window or fixation effect is unreported, and the exact A01470-1 paraffin-section caption does not state a fixative (A01470-1 caption). Heat retrieval in citrate buffer at pH 6 for 20 minutes was used in that mouse brain example; whether ESRRG staining depends on retrieval is unreported (A01470-1 caption). The supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC; kidney sections need background checks for endogenous biotin and peroxidase with SABC/DAB detection (A01470-1 caption; standard IHC practice).
HPA tissue IHC evidence for ESRRG
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — 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 |
|---|---|---|---|---|
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ESRRG IHC Tips
Troubleshoot ESRRG staining in paraffin sections by checking retrieval, compartment, antibody specificity, and whether the measured signal fits the tissue evidence.
How should I retrieve ESRRG when nuclear staining is weak?
Start with heat-mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01470-1). The illustrated paraffin-section workflow then used 10% goat serum and 1 µg/mL primary antibody overnight at 4°C (datasheet A01470-1). If staining remains weak, compare retrieval duration on matched sections while holding antibody concentration and detection constant (standard IHC practice). Evaluate whether signal improves in nuclei, since ESRRG is assigned to the nucleus and nucleoplasm (UniProt P62508; HPA subcellular). Excessive retrieval can damage morphology, so assess tissue preservation alongside signal (standard IHC practice).
Could fixation explain inconsistent ESRRG staining across my paraffin sections?
The selected ESRRG paraffin-section caption does not state a fixative, so target-specific fixation sensitivity is unknown (datasheet A01470-1). Record the fixative, fixation duration, tissue thickness, and processing history for each section before comparing staining (standard IHC practice). Keep those variables consistent across experimental and control sections, then apply the documented citrate pH 6 retrieval for 20 minutes (datasheet A01470-1). If archived blocks differ in processing, run matched positive and negative staining controls with each batch and interpret intensity cautiously (standard IHC practice). Do not assign weak staining to an ESRRG-specific fixation effect without a controlled comparison.
Should I accept mainly cytoplasmic ESRRG staining in my sections?
Prioritise nuclear staining when assessing ESRRG because UniProt assigns it to the nucleus and HPA reports approved nucleoplasmic localisation (UniProt P62508; HPA subcellular). HPA tissue IHC instead describes mainly cytoplasmic staining in a few tissues and rates its staining reliability uncertain because antibody staining has low consistency with RNA expression (HPA tissue IHC). Compare nuclear and cytoplasmic signal separately within identified cell populations, using the same counterstain and exposure conditions across sections (standard IHC practice). Treat isolated cytoplasmic DAB as provisional until it survives antibody omission, detection-only, and independent specificity checks (standard IHC practice).
Can this antibody distinguish ESRRG isoforms in paraffin sections?
ESRRG has 5 listed isoforms, but the supplied caption gives no antibody epitope or isoform cross-reactivity map (UniProt P62508; datasheet A01470-1). Therefore, a positive chromogenic signal cannot identify a particular isoform from this evidence alone. Check the antibody’s mapped immunogen against isoform sequences before making an isoform-specific claim, and validate any proposed distinction with an independent assay (standard antibody-validation practice). The listed ligand-binding domain spans residues 233–457, and serine 45 is a recorded modified residue; neither fact establishes which epitope this antibody recognises (UniProt P62508). Score staining as ESRRG-associated until specificity is established.
How can IF help resolve ambiguous ESRRG staining seen by chromogenic IHC?
Use IF on matched material to assess whether ESRRG overlaps a marker for the expected cell population, such as tubular cells when investigating the kidney signal reported by HPA (HPA tissue IHC). Choose a fluorophore whose emission is well separated from the specimen’s autofluorescence, and include single-colour and no-primary controls to check bleed-through and background (standard IF practice). For the expected nuclear or nucleoplasmic signal, optimise permeabilisation to give the antibody access to intracellular epitopes; ESRRG has no transmembrane segment, while this antibody’s epitope is unspecified (UniProt P62508; HPA subcellular; datasheet A01470-1). Compare the IF compartment with the DAB pattern, treating discordance as a reason for further validation.
How do I reduce diffuse DAB background without losing ESRRG signal?
Begin with the caption’s 10% goat-serum block and 1 µg/mL primary antibody overnight at 4°C as a documented starting point (datasheet A01470-1). Include a no-primary section and a detection-only control to distinguish primary-dependent staining from secondary or chromogen background (standard IHC practice). Because the illustrated method uses a biotinylated secondary, streptavidin–biotin complex, and DAB, assess endogenous biotin and peroxidase contributions where relevant (datasheet A01470-1; standard IHC practice). Optimise blocking, washing, and primary concentration one variable at a time while checking that expected nuclear signal remains interpretable (UniProt P62508; standard IHC practice).
What should I quantify when ESRRG staining varies among cell populations? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because ESRRG is assigned to the nucleus while HPA tissue IHC reports a conflicting mainly cytoplasmic pattern (UniProt P62508; HPA tissue IHC). For chromogenic sections, report the percentage of positive nuclei and an intensity-weighted H-score, or positive-cell density per mm² when cell abundance itself is the endpoint (standard IHC quantification practice). Normalise each measure to the number of evaluable cells or viable tissue area in the same annotated compartment (standard IHC quantification practice). Apply one staining threshold across matched batches, and report cytoplasmic staining separately rather than combining it with the nuclear score.
When is an ESRRG-positive section convincing rather than artefactual?
A convincing result shows reproducible signal in an identified cell population and a plausible nuclear compartment, consistent with the UniProt nuclear and HPA nucleoplasmic assignments (UniProt P62508; HPA subcellular). Kidney tubular cells and stomach glandular cells have medium HPA tissue-IHC staining, but that tissue profile carries an uncertain reliability rating (HPA tissue IHC). Review edge staining, necrotic regions, endogenous peroxidase signal, and staining in no-primary controls before accepting brown DAB deposits as specific (standard IHC practice). Cytoplasmic-only staining or signal in unexpected cells needs independent antibody or orthogonal evidence, especially given the reported disagreement between HPA antibody staining and RNA expression (HPA tissue IHC).
Boster reagents
Best ESRRG / Estrogen-related receptor gamma IHC Antibodies
A01470-1 has IHC images from paraffin sections of mouse brain, rat cardiac muscle and human stomach, plus an IF/ICC image from A549 cells (catalog image captions).
Real IHC data
Anti-strogen Related Receptor gamma/ESRRG Antibody ®
Cat # A01470-1
A01470-1 will render with its mouse brain paraffin-section IHC figure (catalog IHC image caption). Its other captions show IHC in rat cardiac muscle and human stomach paraffin sections and IF/ICC in A549 cells (catalog image captions).
Which to pick: Choose A01470-1 for paraffin-section IHC, supported by its own IHC captions; the fixative is unreported (catalog IHC image captions). Choose the same SKU for IF/ICC based on its application list and A549 image, or for human, mouse and rat work based on its stated reactivity (catalog applications; catalog IF image caption; catalog reactivity). Clonality is unreported (catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P62508 (ERR3_HUMAN, Estrogen-related receptor gamma).
- Human Protein Atlas. ESRRG tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. ESRRG subcellular location (ICC-IF): Localized to the nucleoplasm..
- Human Protein Atlas. ESRRG antibody validation summary (1 antibodies).
- Esrrg functions in early branch generation of the ureteric bud and is essential for normal development of the renal papilla. Human molecular genetics 2011 — PMC3033182.
- Bilateral renal agenesis/hypoplasia/dysplasia (BRAHD): postmortem analysis of 45 cases with breakpoint mapping of two de novo translocations. PloS one 2010 — PMC2928268.
- Deciphering ERR family genes as prognostic and immunological biomarkers through pan-cancer analysis with validation in gallbladder cancer. Frontiers in oncology 2025 — PMC12066295.
- Nucleolar stress in C9orf72 and sporadic ALS spinal motor neurons precedes TDP-43 mislocalization. Acta neuropathologica communications 2021 — PMC7885352.
- PubMed PMID:10072763 — UniProt-cited evidence.
- PubMed PMID:10048485 — UniProt-cited evidence.
- PubMed PMID:9676434 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ESRRG (Estrogen-related receptor gamma). Expected localisation Mainly cytoplasmic in tissue IHC; annotated nuclear (HPA tissue IHC; UniProt). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A01470-1). Validated IHC antibody: A01470-1 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Stomach (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A01470-1); 2. Fixation: Image fixative and duration unreported (datasheet A01470-1); 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: Citrate pH 6 , 20 min (datasheet A01470-1); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01470-1); 8. Primary antibody: Rabbit anti-ESRRG, 0.5-1μg/ml (datasheet A01470-1); 9. Primary incubation: Overnight at 4 °C (datasheet A01470-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01470-1); Caveat: Staining–RNA discordance makes specificity uncertain (HPA tissue IHC); Isoform / epitope: 5 isoforms; verify epitope coverage (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P62508, HPA tissue IHC · Uncertain, HPA ICC-IF · nucleoplasm., Datasheet A01470-1, PMC3033182, PMC12066295. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ESRRG/immunohistochemistry-ihc-guide.