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EDNRA / Endothelin-1 receptor · IHC design guide
Design Immunohistochemistry for EDNRA
Plan chromogenic EDNRA IHC in paraffin sections using 0.5–1 μg/ml of the catalog antibody (datasheet A01828-4). Compare the observed cytoplasmic tissue pattern (HPA tissue IHC) with EDNRA’s annotated membrane location (UniProt), and interpret staining cautiously given uncertain IHC reliability (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 | Tissue cytoplasmic (HPA tissue IHC); molecular 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 fixation consistent across paraffin sections (standard IHC practice; not target-specific) | |
| Caveat | Low staining–RNA concordance; IHC reliability uncertain (HPA tissue IHC) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | 5 isoforms; check extracellular vs cytoplasmic epitope coverage (UniProt) |
Section 1
Recommended EDNRA IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A01828-4) is accompanied by 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 heat-mediated EDTA pH 8.0 retrieval for the catalog antibody (datasheet: A01828-4). Use the reported citric acid conditions when reproducing the carotid artery study (PMC8810813).
Section 2
What Is the Expected EDNRA Staining Pattern?
EDNRA is a seven-transmembrane receptor assigned to the cell membrane (UniProt P25101 topology). In tissue IHC, HPA reports general cytoplasmic staining, including endothelial cells, and medium staining in selected glandular and neural cells (HPA tissue IHC). Treat that appearance as provisional: HPA rates the tissue staining Uncertain because antibody staining and RNA expression show low consistency (HPA tissue IHC).
| Membrane-associated staining in glandular cells of adrenal gland or appendix, with some cytoplasmic signal. | This is plausible EDNRA staining: both cell populations have Medium HPA tissue IHC staining, while UniProt assigns EDNRA to the cell membrane (HPA tissue IHC; UniProt P25101). Cytoplasmic signal needs caution because HPA describes general cytoplasmic tissue staining, but its tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Predominantly nuclear staining, without a convincing membrane-associated component. | A nuclear pattern conflicts with UniProt’s cell-membrane assignment and HPA ICC-IF’s supported plasma-membrane location (UniProt P25101; HPA subcellular). First assess it as possible nonspecific staining or a detection artefact (general IHC practice). HPA’s uncertain cytosol call does not support nuclear localisation (HPA subcellular). |
| Strong staining in a cell population recorded as Not detected, such as liver cholangiocytes. | That result disagrees with the listed HPA tissue observation; adipose-tissue adipocytes and splenic red-pulp cells are also listed as Not detected (HPA tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic activity and check controls (general IHC practice). A discordant HPA result alone cannot prove absence of EDNRA because tissue IHC reliability is Uncertain (HPA tissue IHC). |
| Uniform colour across cells and extracellular areas, obscuring cell boundaries. | This distribution offers little compartment or cell-type information; diffuse signal can reflect nonspecific binding, incomplete blocking, or detection background (general IHC practice). It cannot resolve the difference between UniProt’s membrane assignment and HPA’s general cytoplasmic tissue profile (UniProt P25101; HPA tissue IHC). |
| No staining in adrenal-gland or appendix glandular cells that were selected as positive comparators. | HPA records Medium staining in these cells, so a blank comparator calls for a run-level check (HPA tissue IHC). Review antibody working conditions, retrieval and detection controls using the antibody’s own validated IHC-P instructions (general IHC practice). The HPA profile is Uncertain, so one negative slide is not decisive evidence that EDNRA is absent (HPA tissue IHC). |
💡Expected EDNRA appearanceCall a slide provisionally positive when selected HPA Medium cells, such as adrenal-gland or appendix glandular cells, show cell-associated staining with a plausible membrane component; isolated nuclear colour or uniform background is suspect (HPA tissue IHC; UniProt P25101; general IHC practice).
| Membrane topology and epitope position (UniProt P25101 topology) | EDNRA has seven membrane-spanning segments and extracellular and cytoplasmic regions (UniProt P25101). Interpret an antibody’s pattern against its stated immunogen when that information is available; the supplied record does not identify an antibody epitope (general IHC practice). |
| Tissue profile and its reliability (HPA tissue IHC) | HPA lists Medium staining in several specified cell populations, yet rates tissue IHC Uncertain because staining and RNA data have low consistency (HPA tissue IHC). Use those populations as provisional comparators and assess cell identity and compartment together (general IHC practice). |
| Subcellular evidence across applications (HPA subcellular; UniProt P25101) | UniProt assigns cell membrane; HPA ICC-IF supports plasma membrane and marks cytosol uncertain (UniProt P25101; HPA subcellular). HPA’s tissue IHC profile is generally cytoplasmic, including endothelial cells, so ICC-IF localisation cannot certify a cytoplasmic chromogenic IHC signal (HPA tissue IHC; HPA subcellular). |
| Isoforms and processing (UniProt P25101) | UniProt lists five isoforms, a signal peptide at residues 1–20, and a mature chain at 21–427 (UniProt P25101). Isoform recognition depends on the antibody epitope; this record supplies no antibody-specific coverage, so avoid assigning a stained cell to an isoform (general IHC practice). |
| Glycosylation annotations (UniProt P25101) | Two glycosylation sites are annotated at residues 29 and 62 (UniProt P25101). Their presence does not establish an IHC retrieval requirement or a target-specific fixation effect; those effects are unreported in the supplied sources (UniProt P25101; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparator is blank while the rest of the slide appears technically intact. | Antibody conditions, retrieval or detection may need review (general IHC practice); HPA Medium staining is an uncertain expectation (HPA tissue IHC). | Check the antibody’s validated IHC-P conditions and run controls, then compare adrenal-gland or appendix glandular cells again (general IHC practice; HPA tissue IHC). Do not infer EDNRA absence from this result alone (HPA tissue IHC: Uncertain). |
| Every tissue shows broad, similar chromogenic colour. | Nonspecific binding or endogenous detection activity may obscure cell-specific staining (general IHC practice). | Inspect a no-primary control and the relevant enzyme-blocking step; reassess localisation after background is controlled (general IHC practice). A broad signal does not match a cell-resolved interpretation of the HPA observations (HPA tissue IHC). |
| Colour is concentrated in nuclei rather than along cell borders. | The compartment conflicts with EDNRA’s membrane assignment and HPA’s supported plasma-membrane ICC-IF location (UniProt P25101; HPA subcellular). | Review the no-primary control, chromogen distribution and antibody conditions before scoring (general IHC practice). Keep nuclear staining separate from any plausible cytoplasmic signal, which HPA rates uncertain in ICC-IF (HPA subcellular). |
| A presumed negative population stains strongly. | Cross-reactivity or endogenous detection activity is possible (general IHC practice); the chosen population may also be a poor comparator if its identity is unclear. | Confirm cell identity and compare with the precise HPA entry, such as liver cholangiocytes listed as Not detected (HPA tissue IHC). Use a no-primary control and interpret disagreement cautiously because HPA tissue IHC reliability is Uncertain (general IHC practice; HPA tissue IHC). |
| Vascular staining appears inconsistent between specimens. | HPA describes endothelial-cell cytoplasmic staining, while UniProt reports no expression in umbilical vein endothelial cells; these statements concern different evidence and cell contexts (HPA tissue IHC; UniProt P25101). | Record vessel and cell identity before comparing sections; do not treat every endothelial population as an interchangeable positive control (HPA tissue IHC; UniProt P25101). Evaluate chromogenic background with run controls (general IHC practice). |
| IF/ICC: should a cytosolic fluorescent signal count as EDNRA positive? | HPA ICC-IF supports plasma membrane but calls cytosol uncertain; its tissue IHC cytoplasmic profile is also rated Uncertain (HPA subcellular; HPA tissue IHC). | Give greater weight to a cell-associated plasma-membrane pattern and assess controls before calling cytosolic fluorescence specific (HPA subcellular; general IF practice). Use the separate IF/ICC guide for that application; chromogenic IHC observations alone do not validate an IF result (HPA tissue IHC; general IF practice). |
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 assess staining in its glandular cells (HPA: Medium in adrenal gland glandular cells). Use liver cholangiocytes as the negative tissue (HPA: Not detected in liver cholangiocytes); on the adrenal slide, use adjacent cells without specific staining to judge background, without assuming a particular cell type is 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 concentration-matched rabbit IgG isotype control, and EDNRA-knockout tissue as a biological negative where available (selected-SKU caption: rabbit primary; standard IHC practice). For the caption’s SABC–DAB detection, block endogenous peroxidase and assess endogenous biotin background on the adrenal section (selected-SKU caption: SABC–DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected-SKU paraffin-section caption does not state a fixative. The reported IHC procedure uses heat retrieval in EDTA at pH 8.0 and 1 µg/ml primary antibody overnight at 4°C; retrieval dependence and whether frozen sections or IF are easier have not been established (selected-SKU caption). Adrenal sections require attention to nonspecific chromogenic background when using the reported SABC–DAB method (selected-SKU caption; 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 scoring against the receptor’s membrane localization (UniProt P25101).
Which retrieval conditions should I try first for EDNRA in paraffin sections?
Use heat mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A01828-4). The selected tissue image used this retrieval before incubation with 1 µg/ml primary antibody overnight at 4°C (caption A01828-4). If staining is weak, compare retrieval duration across matched sections while keeping antibody concentration and detection conditions fixed (standard IHC practice). Excessive heat can damage morphology, so assess tissue integrity alongside staining rather than choosing the strongest chromogen signal alone (standard IHC practice). Interpret any gain against a negative control and the expected membrane distribution of this seven transmembrane receptor (UniProt P25101 topology).
How should I troubleshoot fixation when EDNRA staining varies between paraffin samples?
Target specific fixation sensitivity is unknown because the selected paraffin tissue caption does not state its fixative (caption A01828-4). Record fixative, fixation duration, tissue thickness, and processing history for each sample before comparing signal across blocks (standard IHC practice). Where material permits, compare similarly processed sections with the same pH 8.0 EDTA retrieval and 1 µg/ml antibody condition (caption A01828-4). Check whether weak staining coincides with poor morphology or uneven staining across the section, which can reveal processing variation (standard IHC practice). Do not infer a fixation effect from tissue expression patterns or receptor topology alone (HPA tissue IHC; UniProt P25101 topology).
What EDNRA staining pattern should I expect in tissue sections?
Prioritize staining at cell borders because EDNRA is annotated at the cell membrane and contains 7 transmembrane segments (UniProt P25101). Cytoplasmic staining needs caution: the subcellular atlas supports plasma membrane localization but labels cytosol localization uncertain (HPA subcellular). Tissue IHC reports general cytoplasmic expression, including endothelial cells, yet assigns the staining uncertain reliability because it poorly matches RNA data (HPA tissue IHC). Compare staining within defined cell populations; adrenal glandular cells and bronchial respiratory epithelial cells have reported medium staining (HPA tissue IHC). Review membrane detail at high magnification and compare adjacent negative control sections before assigning diffuse cytoplasmic signal to EDNRA (standard IHC practice).
Could isoforms or epitope position explain inconsistent EDNRA staining?
EDNRA has 5 annotated isoforms, so confirm the catalog antibody’s immunogen sequence before claiming that a stain detects every isoform (UniProt P25101; standard IHC practice). The supplied product caption does not identify its epitope, leaving isoform coverage and epitope orientation unresolved for this antibody (caption A01828-4). EDNRA has extracellular residues 21–80, cytoplasmic residues 373–427, and seven membrane segments that constrain where an epitope can lie (UniProt P25101 topology). Glycosylation sites at residues 29 and 62 warrant consideration if a mapped epitope overlaps that region, without establishing an effect on this antibody (UniProt P25101). Compare the mapped immunogen with isoform sequences before interpreting discordant tissue staining as isoform selective (standard IHC practice).
How can I check an EDNRA IHC finding with multiplex immunofluorescence?
Use immunofluorescence as a separate confirmation experiment, because the selected product caption describes chromogenic paraffin IHC rather than an IF protocol (caption A01828-4). Pair EDNRA with a marker for the population being assessed, such as an endothelial marker when testing the reported endothelial staining (HPA tissue IHC; standard IF practice). Choose a far red EDNRA fluorophore if the specimen has strong shorter wavelength autofluorescence, and include single stain controls for spectral bleed through (standard IF practice). If the antibody epitope is cytoplasmic, test permeabilisation; an extracellular epitope may be accessible without it, but this antibody’s epitope is unspecified (UniProt P25101 topology; caption A01828-4). Compare membrane signal with matched negative controls before relating IF intensity to chromogenic IHC (HPA subcellular; standard IF practice).
How do I reduce diffuse or vascular background in EDNRA chromogenic IHC?
The selected image used 10% goat serum blocking, a biotinylated secondary, streptavidin biotin complex, and DAB (caption A01828-4). For excess chromogen, inspect a no primary control to separate secondary or detection background from antibody dependent staining (standard IHC practice). Include a peroxidase block before DAB development and consider endogenous biotin when using the caption’s biotin based detection system (standard IHC practice; caption A01828-4). Titrate primary antibody around the caption’s 1 µg/ml condition and use consistent washes across matched sections (caption A01828-4; standard IHC practice). Judge reductions in background against preserved cell border signal, the better supported EDNRA location (HPA subcellular; UniProt P25101).
How should I score EDNRA staining across tissue groups? ⚠ ANSWER MARKED FOR VERIFICATION
Define the target cell population and viable tissue area before scoring, then apply one chromogen threshold across comparably processed sections (standard IHC practice). Report an H-score from staining intensity and percentage of positive cells, or report positive cells per mm² when cell counts suit the question (standard IHC practice). Normalize percentages to all evaluable cells of the same type and density to viable tissue area, excluding folds and necrotic regions (standard IHC practice). Record membrane associated and diffuse cytoplasmic staining separately because membrane localization is supported while cytosol localization is uncertain (HPA subcellular). Treat comparisons as assay dependent until controls establish specificity, given the uncertain tissue IHC reliability reported for EDNRA (HPA tissue IHC).
How can I distinguish genuine EDNRA staining from artefact?
Give greatest weight to reproducible cell border staining in intact cells, consistent with EDNRA’s cell membrane annotation and 7 transmembrane segments (UniProt P25101). Diffuse cytoplasmic color alone is less persuasive because cytosol localization is uncertain and tissue IHC staining has uncertain reliability (HPA subcellular; HPA tissue IHC). Check whether signal concentrates at section edges, folds, or necrotic regions, and compare those areas with intact tissue (standard IHC practice). Use a no primary control to identify endogenous enzyme or detection system staining when interpreting DAB deposits (standard IHC practice). Finally, identify the stained cell type explicitly: the atlas reports medium staining in adrenal glandular cells but no detection in liver cholangiocytes (HPA tissue IHC).
Boster reagents
Best EDNRA / Endothelin-1 receptor IHC Antibodies
One human-reactive anti-EDNRA antibody has IHC data from a paraffin-embedded human lung cancer section (A01828-4 image caption); no IF figure is supplied (catalog: IF images).
Real IHC data
Anti-Endothelin A Receptor/ET-A/EDNRA Antibody ®
Cat # A01828-4
A01828-4 is listed for human IHC (catalog: A01828-4 applications and reactivity). Its IHC figure shows a paraffin-embedded human lung cancer section (A01828-4 image caption).
Which to pick: Choose A01828-4 for human paraffin-section IHC; its figure used EDTA pH 8.0 retrieval and 1 μg/ml primary antibody, but did not report the fixative (A01828-4 image caption). No listed SKU has documented IF/ICC validation (catalog: A01828-4 applications and IF images). No listed SKU supports a cross-species choice because A01828-4 lists human reactivity only (catalog: A01828-4 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 Tissue cytoplasmic (HPA tissue IHC); molecular 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 fixation consistent across paraffin sections (standard IHC practice; not target-specific); Caveat: Low staining–RNA concordance; IHC reliability uncertain (HPA tissue IHC); Isoform / epitope: 5 isoforms; check extracellular vs cytoplasmic epitope coverage (UniProt); Regulation: 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.