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EDNRA / Endothelin-1 receptor · IHC design guide
Design Immunohistochemistry for EDNRA
Plan chromogenic EDNRA IHC on paraffin sections with the IHC-validated antibody at 0.5–1 μg/mL (datasheet A01828-4). Interpret general cytoplasmic staining, including endothelial cells, cautiously because staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
EDNRA Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic in tissue (HPA tissue IHC); membrane (UniProt) | |
| Staining pattern | General cytoplasmic staining, including endothelial cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A01828-4) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+2 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent (standard IHC practice; not target-specific) | |
| Caveat | Low consistency between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Isoform expression varies by tissue (UniProt) | |
| Isoform / epitope | 5 isoforms; check extracellular vs cytoplasmic epitope (UniProt) |
Section 1
Recommended EDNRA IHC & IF Protocols
The catalog antibody's IHC-P protocol (datasheet A01828-4) appears alongside four published EDNRA IHC protocols (PMC9066206; PMC13398129; PMC10526862; PMC8810813).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet A01828-4) |
| Fixation | Image fixative and duration unreported (datasheet A01828-4); 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 A01828-4); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A01828-4) |
| Primary antibody | Rabbit anti-EDNRA, 0.5-1μg/ml (datasheet A01828-4) |
| Primary incubation | Overnight at 4 °C (datasheet A01828-4) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01828-4) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | EDNRA-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: General cytoplasmic expression, endothelial cells including. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A01828-4). A mouse carotid artery study used citric acid retrieval (PMC8810813).
Section 2
What Is the Expected EDNRA Staining Pattern?
EDNRA is a seven-transmembrane cell-membrane receptor (UniProt P25101 topology and subcellular location). In paraffin-section IHC, assess staining in the identified cells: HPA reports medium staining in adrenal glandular cells, bronchial respiratory epithelial cells and cerebellar granular-layer cells (HPA tissue IHC). HPA also describes general cytoplasmic staining, including endothelial cells, but rates its tissue IHC evidence Uncertain because staining and RNA expression have low consistency (HPA tissue IHC).
| Cell borders are stained in an identifiable glandular or respiratory epithelial population. | A membrane-associated pattern fits EDNRA's seven-transmembrane topology (UniProt P25101 topology). Adrenal glandular and bronchial respiratory epithelial cells have medium staining in the HPA tissue record (HPA tissue IHC). Score the named cell population and its staining distribution; a stained field alone does not establish EDNRA specificity because HPA rates tissue IHC Uncertain (HPA tissue IHC). |
| Staining is confined to nuclei, or forms isolated deposits unrelated to cell outlines. | A nuclear-only pattern does not match EDNRA's annotated cell-membrane location (UniProt P25101 subcellular location). Discrete deposits may reflect chromogen precipitate (general IHC practice). Recheck morphology and the detection controls before calling these cells positive. Cytoplasmic staining by itself needs more care: HPA describes it in tissue IHC, while its ICC-IF cytosol assignment is uncertain (HPA tissue IHC; HPA subcellular). |
| Strong staining appears in a cell population expected to be negative in the chosen reference section. | HPA reports cholangiocytes in liver and red-pulp cells in spleen as not detected in its tissue IHC examples (HPA tissue IHC). Review cell identification and a control without primary antibody; unexpected color may arise from antibody cross-reactivity or endogenous detection activity (general IHC practice). An HPA negative example is a comparison point, not proof that every specimen of that tissue must be negative (HPA tissue IHC: Uncertain). |
| Color spreads across the section without a reproducible cell or compartment pattern. | Diffuse staining cannot be assigned confidently to EDNRA's membrane compartment (UniProt P25101 subcellular location). In chromogenic IHC, incomplete blocking, insufficient washing or overly concentrated antibody can increase background (general IHC practice). Compare the no-primary control and inspect whether any signal remains specifically associated with identifiable cells before scoring (general IHC practice). |
| No cells stain in an adrenal or bronchial section chosen as a positive reference. | HPA records medium staining in adrenal glandular and bronchial respiratory epithelial cells (HPA tissue IHC). First confirm that the relevant cells are present, then check the antibody's documented IHC-P conditions and the detection run (general IHC practice). Because HPA tissue IHC reliability is Uncertain, an unstained reference does not by itself establish biological absence or a failed assay (HPA tissue IHC). |
💡Expected EDNRA appearanceCall a result provisionally positive when identifiable adrenal glandular or bronchial respiratory epithelial cells show cell-associated staining, preferably at the membrane and potentially with cytoplasmic signal, at an intensity comparable to HPA's medium examples; isolated nuclear color, precipitate or uniform background is a suspect positive (UniProt P25101 subcellular location; HPA tissue IHC: Medium, Uncertain; general IHC practice).
| Compartment and accessible sequence | EDNRA has 7 membrane-spanning segments and extracellular and cytoplasmic regions (UniProt P25101 topology). The supplied record gives no antibody epitope, so topology alone cannot predict whether a particular retrieval or permeabilisation condition improves this antibody's IHC signal. |
| Tissue and cell selection | HPA's medium examples name specific cells, including cerebellar granular-layer cells, whereas liver cholangiocytes are not detected (HPA tissue IHC). Compare like cell populations rather than assigning one expected intensity to an entire organ; HPA's tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Antibody validation | Both listed antibodies have Uncertain IHC status, HPA013774 and CAB018957 (HPA antibodies). Treat the tissue pattern as a working expectation and seek concordance across morphology and controls; the HPA record does not establish an IHC-enhanced antibody. |
| Isoforms and processing | UniProt lists 5 EDNRA isoforms, a signal peptide at residues 1–20 and a receptor chain at 21–427 (UniProt P25101 isoforms and processing). Without the antibody epitope, these annotations cannot identify which isoforms the stain detects or justify a shedding-based explanation for missing staining. |
| IF/ICC Q: what localization should I expect? | A: HPA supports plasma-membrane localization and marks cytosol uncertain in ICC-IF; images are listed for U-251MG and U2OS (HPA subcellular). That result helps interpret compartment, but it does not supply a paraffin IHC protocol or establish that cytoplasmic tissue staining is specific. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive-reference cells are absent from the section. | The sampled field may lack the cell population used for comparison (general IHC practice). | Locate the relevant glandular or epithelial cells before judging the run; HPA's medium examples refer to adrenal glandular and bronchial respiratory epithelial cells specifically (HPA tissue IHC). |
| Reference cells are present but the slide has no signal. | A missed detection step or unsuitable antibody conditions are possible (general IHC practice); HPA reference staining is uncertain (HPA tissue IHC). | Check the catalog antibody's documented IHC-P procedure, reagent performance and detection controls (general IHC practice). Do not infer EDNRA-specific fixation sensitivity from this result; none is supplied. |
| The no-primary control develops color. | Endogenous detection activity or nonspecific reagent deposition may contribute to chromogenic signal (general IHC practice). | Address the detected background with the appropriate blocking and washing controls, then repeat the comparison (general IHC practice). Avoid assigning the control's color to EDNRA. |
| Staining is diffuse or obscures cell boundaries. | High background can follow insufficient blocking or washing, or an overly concentrated primary antibody (general IHC practice). | Optimize those general IHC conditions against a no-primary control and score only resolved cellular signal (general IHC practice). No EDNRA-specific dilution or retrieval setting is provided here. |
| Only nuclei or extracellular specks look positive. | Those appearances conflict with EDNRA's annotated cell-membrane location; specks may be precipitate (UniProt P25101 subcellular location; general IHC practice). | Inspect adjacent morphology and repeat detection controls before accepting the stain; seek a reproducible cell-associated distribution in the HPA-listed populations (HPA tissue IHC; general IHC practice). |
| Cytoplasmic staining appears without a clear membrane rim. | HPA describes general cytoplasmic tissue staining, but its ICC-IF cytosol localization is uncertain and tissue IHC reliability is uncertain (HPA tissue IHC; HPA subcellular). | Record the compartment separately from intensity, compare the named reference cells and controls, and report the pattern cautiously (general IHC practice; HPA tissue IHC: Uncertain). |
Experiment evidence
IHC & IF images showing EDNRA staining
A01828-4 · IHCIHC analysis of Endothelin A Receptor/ET-A/ EDNRA using anti-Endothelin A Receptor/ET-A/ EDNRA antibody (A01828-4). Endothelin A Receptor/ET
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000151617-EDNRA/tissue/cerebral+cortex#imid_4806444
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000151617-EDNRA/tissue/colon#imid_4806378
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000151617-EDNRA/tissue/liver#imid_4806379
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000151617-EDNRA/tissue/kidney#imid_4806388
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB018957; green = EDNRA, blue = nucleus, red = microtubules.
HPA · ICC-IF U-251MGHuman Protein Atlas immunofluorescence, U-251MG, antibody CAB018957; green = EDNRA, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for EDNRA IHC & IF
🧪Run adrenal gland first and score glandular cells for EDNRA staining (HPA: Medium in adrenal gland glandular cells). Use liver cholangiocytes as the negative comparator (HPA: Not detected in liver cholangiocytes); on the adrenal slide, use cells outside the glandular compartment to assess background without assuming they are EDNRA-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 EDNRA in U-251MG, U2OS, with annotated localisation: Plasma membrane (supported), Cytosol (uncertain) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a nonimmune rabbit IgG isotype control matched to the primary’s concentration and clonality where applicable (caption: rabbit primary), and a verified EDNRA-knockout biological comparator. For chromogenic detection, block endogenous peroxidase and check for endogenous biotin background in the adrenal section (caption: streptavidin–biotin detection with DAB).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A01828-4 tissue-IHC caption does not state the fixative. That paraffin-section example uses heat-mediated EDTA retrieval at pH 8.0 (caption), but whether EDNRA staining depends on that retrieval condition is unreported; neither frozen sections nor IF is established here as easier. In adrenal gland, compare DAB staining with the no-primary section to distinguish signal from any endogenous pigment (standard IHC practice).
HPA tissue IHC evidence for EDNRA
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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | Glial cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced EDNRA IHC Tips
Troubleshoot EDNRA staining in paraffin sections by checking retrieval, compartment, controls, and cell-specific scoring before interpreting chromogenic signal.
Which retrieval conditions should I try first for EDNRA IHC?
Start with heat-mediated retrieval in EDTA pH 8.0 for paraffin sections (datasheet A01828-4). The selected tissue image used that retrieval before incubation with 1 μg/ml catalog antibody overnight at 4°C (caption A01828-4). Keep section thickness, heating time, and cooling conditions consistent across test and control sections, then compare membrane-associated staining and tissue preservation (standard IHC practice; UniProt P25101 subcellular location). If staining remains weak, test citrate pH 6.0 as a fallback on matched sections, changing only the retrieval condition and recording any rise in diffuse background (standard IHC practice).
How should I troubleshoot fixation when EDNRA staining varies between sections?
The selected EDNRA image documents a paraffin section but does not state its fixative, so target-specific fixation sensitivity is unknown (caption A01828-4). Record fixative, fixation duration, specimen thickness, and processing history for each case; compare sections processed together before attributing intensity differences to EDNRA abundance (standard IHC practice). If historical fixation details are missing, run the catalog antibody and a no-primary control on matched sections using EDTA pH 8.0 retrieval (datasheet A01828-4; standard IHC practice). Assess morphology and staining across the section, because uneven preservation can distort chromogenic comparisons even when the antibody incubation is identical (standard IHC practice).
What staining pattern is plausible for EDNRA in tissue?
Prioritise membrane-associated signal because EDNRA is a 7-transmembrane receptor annotated at the cell membrane (UniProt P25101 topology and subcellular location). Some intracellular signal may be plausible, but the HPA subcellular record supports plasma membrane localisation while rating cytosol localisation uncertain (HPA subcellular). Interpret diffuse cytoplasmic DAB cautiously: HPA describes general cytoplasmic tissue staining, yet rates its tissue-IHC reliability uncertain because staining and RNA data have low consistency (HPA tissue IHC). Compare cell borders, cytoplasm, and nearby unstained structures at the same exposure and magnification; use a no-primary section to assess nonspecific chromogen deposition (standard IHC practice).
Could EDNRA isoforms or epitope position explain inconsistent staining?
EDNRA has 5 annotated isoforms, so check the catalog antibody’s immunogen sequence against the isoform relevant to the specimen before assigning absent staining to absent protein (UniProt P25101 isoforms; standard IHC practice). The mature receptor spans residues 21–427, with extracellular and cytoplasmic loops plus a cytoplasmic C-terminal region (UniProt P25101 processing and topology). Glycosylation is annotated at residues 29 and 62, and phosphoserine at 425; these annotations alone do not establish antibody recognition of any modified state (UniProt P25101 modifications). If the immunogen is undisclosed, report epitope and isoform coverage as unresolved, and compare matched positive and low-expression controls (standard IHC practice; HPA tissue IHC).
How can I cross-check EDNRA localisation with multiplex IF?
Use IF as a complementary localisation check, pairing EDNRA with a marker for the cell population under study, such as an endothelial marker when testing endothelial staining (HPA tissue IHC; standard IF practice). Choose a fluorophore channel with low tissue autofluorescence, and inspect unstained and single-label controls before interpreting overlap (standard IF practice). Match permeabilisation to the antibody epitope: an extracellular epitope can be tested without detergent, while cytoplasmic loops or the C-terminal region generally require permeabilisation (UniProt P25101 topology; standard IF practice). Compare any membrane signal with chromogenic IHC cautiously, because HPA supports plasma membrane localisation but rates cytosol localisation uncertain (HPA subcellular).
How do I reduce diffuse or vascular background in EDNRA IHC?
First compare the primary-stained section with a no-primary control, and check whether DAB accumulates in damaged areas, section edges, or vascular spaces (standard IHC practice). The selected image used 10% goat serum blocking, a biotinylated secondary, a streptavidin-biotin complex, and DAB development (caption A01828-4). Apply a peroxidase block and assess endogenous biotin when using that detection scheme; these are general chromogenic IHC controls, not evidence of EDNRA expression (standard IHC practice). If diffuse staining persists, titrate primary concentration around the documented 1 μg/ml condition and compare specific membrane-associated signal against background on matched sections (caption A01828-4; standard IHC practice).
What is a defensible way to quantify EDNRA IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and region before scoring, then record the percentage of positive cells and staining intensity using a consistent H-score rubric (standard IHC practice). If studying vessels, report positive cells or stained area per mm² and normalise to vessel count or analysed tissue area, respectively (standard IHC practice). Score membrane-associated and diffuse cytoplasmic signal separately because membrane localisation is supported while cytosol localisation is uncertain (UniProt P25101 subcellular location; HPA subcellular). Exclude folds, necrosis, and section edges using the same rules across cases, and document observer agreement because the HPA tissue-IHC profile has uncertain reliability (standard IHC practice; HPA tissue IHC).
When should I call EDNRA staining a true positive?
Call a finding credible when reproducible signal occurs in morphologically intact cells with a plausible membrane-associated pattern and exceeds matched no-primary background (UniProt P25101 subcellular location; standard IHC practice). Treat isolated nuclear staining, edge-only staining, and DAB deposits over necrosis or endogenous enzyme activity as artefact candidates until controls resolve them (standard IHC practice). Check the identified cell type carefully: HPA reports endothelial cells among stained populations, but labels its tissue-IHC profile uncertain owing to low staining–RNA consistency (HPA tissue IHC). A negative or positive region alone is insufficient proof; compare anatomy, controls, and an independent expression measure where available (standard IHC practice; HPA tissue IHC).
Boster reagents
Best EDNRA / Endothelin-1 receptor IHC Antibodies
The IHC-validated anti-EDNRA antibody has real staining data from human lung cancer paraffin sections; no IF image is provided (catalog: A01828-4 IHC image caption; IF image list).
Real IHC data
Anti-Endothelin A Receptor/ET-A/EDNRA Antibody ®
Cat # A01828-4
A01828-4 is listed for human IHC, and its image shows staining in a paraffin section of human lung cancer tissue (catalog: A01828-4 applications, reactivity, IHC image caption). A01828-4 has no IF image or listed IF/ICC application (catalog: A01828-4 applications, IF image list).
Which to pick: Choose A01828-4 for human tissue IHC: its own image documents a paraffin section, and the catalog lists an IHC dilution of 0.5–1 μg/ml (catalog: A01828-4 IHC image caption, dilution). The caption does not report the fixative (catalog: A01828-4 IHC image caption). No SKU in the payload is listed for IF/ICC or species beyond human, so there is no supported IF/ICC or cross-species pick (catalog: A01828-4 applications, reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P25101 (EDNRA_HUMAN, Endothelin-1 receptor).
- Human Protein Atlas. EDNRA tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. EDNRA subcellular location (ICC-IF): Localized to the plasma membrane and cytosol..
- Human Protein Atlas. EDNRA antibody validation summary (2 antibodies).
- Comprehensive analysis of expression signature and immune microenvironment signature of biomarker Endothelin Receptor Type A in stomach adenocarcinoma. Journal of Cancer 2022 — PMC9066206.
- EDNRA Forms a Positive Feedback Loop with the Hippo/YAP Axis to Drive Triple-Negative Breast Cancer Progression. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13398129.
- EDNRA-Expressing Mesenchymal Cells Are Expanded in Myeloma Interstitial Bone Marrow and Associated with Disease Progression. Cancers 2023 — PMC10526862.
- Andrographolide Promotes Interaction Between Endothelin-Dependent EDNRA/EDNRB and Myocardin-SRF to Regulate Pathological Vascular Remodeling. Frontiers in cardiovascular medicine 2021 — PMC8810813.
- PubMed PMID:1719979 — UniProt-cited evidence.
- PubMed PMID:1659806 — UniProt-cited evidence.
- PubMed PMID:1652463 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for EDNRA (Endothelin-1 receptor). Expected localisation Cytoplasmic in tissue (HPA tissue IHC); membrane (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A01828-4). Validated IHC antibody: A01828-4 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 lung cancer tissue; fixative not specified (datasheet A01828-4); 2. Fixation: Image fixative and duration unreported (datasheet A01828-4); 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 A01828-4); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A01828-4); 8. Primary antibody: Rabbit anti-EDNRA, 0.5-1μg/ml (datasheet A01828-4); 9. Primary incubation: Overnight at 4 °C (datasheet A01828-4). Watch for: Fixation: Keep paraffin-section fixation consistent (standard IHC practice; not target-specific); Caveat: Low consistency between staining and RNA expression (HPA tissue IHC); Isoform / epitope: 5 isoforms; check extracellular vs cytoplasmic epitope (UniProt); Regulation: Isoform expression varies by tissue (UniProt). Evidence: UniProt P25101, HPA tissue IHC · Uncertain, HPA ICC-IF · plasma membrane and cytosol., Datasheet A01828-4, PMC9066206, PMC13398129. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/EDNRA/immunohistochemistry-ihc-guide.