Big Endothelin-1 ELISA Assay


The Big Endothelin-1 ELISA Assay Kit is for the determination of Big Endothelin-1 in serum and plasma. The Eagle Biosciences Big Endothelin-1 ELISA Assay Kit is for research use only and not to be used in diagnostic procedures.

SKU: BI-20082H Categories: , ,

Big Endothelin-1 ELISA Assay

Big Endothelin-1 ELISA Assay Developed and Manufactured in Austria by Biomedica

Size: 1×96 wells
Sensitivity: 0.02 pmol/ml
Dynamic Range: 0.1 – 3 pmol/L
Incubation Time: 5.5 hours
Sample Type: serum, plasma
Sample Size: 50 µL
Alternative Names: Big ET-1 ELISA, Human Big Endothelin-1 ELISA, Human Big ET-1 ELISA
For Research Use Only

Controls Included

Values From Apparently Healthy Individuals (n=41):
Serum: Median: 0.09 pmol/l
Each laboratory should establish its own reference range for the samples under investigation.
Do not change sample type during studies.

Assay Principle

The Big Endothelin-1 ELISA Assay kit is a sandwich enzyme immunoassay for the quantitative determination of Big Endothelin-1 in human serum and plasma samples. In a first step, standard/control/sample and detection antibody (monoclonal mouse anti- human Big Endothelin-1 antibody) are pipetted into the wells of the microtiter strips, which are pre-coated with polyclonal sheep anti-human Big Endothelin-1 antibody. Big Endothelin-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. After a washing step, which removes all non-specifically bound and unbound material, the conjugate (streptavidin-HRP) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (tetramethylbenzidine, TMB) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of Big Endothelin-1. This color change is detectable with a standard microtiter plate reader. The concentration of Big Endothelin-1 in the sample is determined directly from the dose response curve.
Areas of Interest

  • prognostic value in heart failure and acute myocardial infarction
  • renal insufficiency
  • during and after graft rejection
  • atherosclerosis
  • pulmonary hypertension and scleroderma

Related Products

Mouse Anti Endothelin Receptor A IgG Antibody ELISA Assay Kit
Endostatin ELISA Assay Kit
NT-proBNP ELISA Assay Kit
Learn more about Endothelin at Eagle Biosciences Biomarker Spotlight page dedicated to Endothelin here: EagleBio Spotlight: Endothelin

Additional Information

Assay Background

Big Endothelin-1 (BigET) is a peptide of 38 amino acids and is the precursor of Endothelin-1 (ET), represented by amino acids 1-21. ET is a potent vasoconstrictor and is produced by vascular endothelial cells. Accordingly it has a wide tissue distribution. The cleavage of BigET by Endothelin Converting Enzyme (ECE) leads to ET and to a C-terminal fragment. Both BigET and ET are strong independent predictors of survival in patients with congestive heart failure, and identify a population with a very high risk mortality. The half-life of ET (1-21) in plasma is less than one minute, whereas clearance of BigET is much slower. BigET can therefore be determined more easily.

Assay Principle

  1. All reagents and samples must be at room temperature (18-26°C) before use in the assay.
  2. Mark position for BLANK/STD (Standards)/SAMPLE/CTRL (Control) on the supplied protocol sheet.
  3. Take microtiter strips out of the aluminum bag, take a minimum of one well as Blank. Store unused strips with desiccant at 2-8°C in the aluminum bag. Strips are stable until expiry date stated on the label.
  4. Add 50 µl STD/SAMPLE/CTRL (Standard/Sample/Control) in duplicate into respective well, except blank.
  5. Add 200 µl AB (Detection antibody) into each well, except blank, swirl gently.
  6. Cover tightly and incubate overnight (16-24 hours) at room temperature (18-26°C).
  7. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer), remove remaining WASHBUF by hitting plate against paper towel after the latest wash.
  8. Add 200 µl CONJ (Conjugate) into each well.
  9. Cover tightly and incubate 1 hour at room temperature (18-26°C).
  10. Aspirate and wash wells 5x with 300 µl diluted WASHBUF (Wash buffer), remove remaining WASHBUF by hitting plate against paper towel after the last wash.
  11. Add 200 µl SUB (Substrate) into each well.
  12. Incubate for 30 minutes at room temperature (18-26°C) in the dark.
  13. Add 50 µl STOP (Stop solution) into each well, shake well.
  14. Measure absorbance immediately at 450 nm with reference 630 nm, if available.

Package Inserts

Please note: All documents above are for reference use only and should not be used in place of the documents included with this physical product. If digital copies are needed, please contact us.

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