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#27169 Endothelin-3 Assay Kit - IBL (Product On Request)

  • #27169 Endothelin-3 ELISA Kit
  • #27169 Endothelin-3 ELISA Kit
Intended Use：
Research reagents
Measuring Method：
ELISA
Sample Types：
Human
Measuring Samples：
EDTA-plasma, Serum, Tissue extract, Cell culture supernatant
Measurement Range：
0.78 ~ 50 pg/mL
Package Size1：
96 Well

※ The product indicated as "Research reagents" in the column Intended Use cannot be used
　 for diagnostic nor any medical purpose.
※ The datasheet listed on this page is sample only. Please refer to the datasheet
　 enclosed in the product purchased before use.

Product Overview

Product Overview

Product Code 27169
Product Name Endothelin-3 Assay Kit - IBL (Product On Request)
Intended Use Research reagents
Measuring Method ELISA
Conjugate HRP
Species Human
Measuring Samples EDTA-plasma, Serum, Tissue extract, Cell culture supernatant
Measurement Range 0.78 ~ 50 pg/mL
Primary Reaction Overnight at 2 - 8 ℃
Secondary Reaction 30 minutes at 2 - 8 ℃
Sensitivity 0.36 pg/mL
Specificity Compound Cross Reactivity
Endothelin-3 100.0%
Endothelin-1 7.50%
Endothelin-1 (1-31) ≦0.1%
Endothelin-2 (1-31) ≦0.1%
Human Big Endothelin-1 ≦0.1%
Rat Big Endothelin-1 0.60%
Storage Condition 2 - 8 ℃
Poisonous and Deleterious Substances Not Applicable
Cartagena Not Applicable
Measuring Service Not Available
Package Size 1 96 Well

Product Description

Product Description

Endothelins (ETs) are isopeptides produced by vascular endothelium having potent vasoconstriction activity. The peptides are encoded by three separate genes and processed to yield 39 residue Big Endothelin (Big ET) molecule, which are further processed to the 21 amino acid sequences termed Endothelin-1 (ET-1), Endothelin-2 (ET-2) and Endothelin-3 (ET-3). All of members of the endothelin family contain two essential disulfide bridges and size conserved amino acid residues at the C-terminus. The ETs are produced by a variety of issues in vivo, including lung, kidney, brain, pituitary and placenta.

※ Regarding the method of extruction, please refer to the section "Attention for sample handling" under "PRECAUTION FOR INTENDED USE AND/OR HANDLING" on page 2 of the datasheet.

Please refer to the following publications for sample pre-treatment and the extraction process.

Plasma
A sandwich-type enzyme immunoassay to detect immunoreactive big-endothelin-1 in plasma. N Suzuki et al. J Immunol Methods
. 1990 Mar 9;127(2):165-70.
A sensitive sandwich-enzyme immunoassay for human endothelin. N Suzuki et al. J Immunol Methods. 1989 Mar 31;118(2):245-50
Serum
Serum endothelin-1 concentrations and cold provocation in primary Raynaud's phenomenon. M R Zamora et al. Clinical Trial Lancet. 1990 Nov 10;336(8724):1144-7.
Granulocyte
Specific sandwich-type enzyme immunoassays for smooth muscle constricting novel 31-amino acid endothelins. N Okishima et al. Biochem Biophys Res Commun. 1999 Mar 5;256(1):1-5.

FAQ

FAQ

  • Question
    Q.Why extraction and condensation process using Sep-Pack is required for measuring Endothelin-3 (ET-3) blood samples?
    Is the process essential for the assay?
  • Answer
    A.Measuring ET-3 in blood sample is quite difficult due to very low concentration of ET-3 (reference approx. 0.48pg/mL in normal subject blood sample).
    The extraction and condensation process using Sep-Pack is essential for the assay.
    The assay will be failed without the process.
    The ET-3 in blood sample become much lower than the original concentration after the process and it depends on the collection rate.
    Therefore, it is recommended to prepare larger volume 5mL-10mL of blood sample for ET-3 measurement.
    Please refer to "Attention for sample handling" on page 2 of the datasheet for additional information.
  • Question
    Q.Which region are recognized by the antibodies used by this kit?
  • Answer
    A.The capture antibody (Anti-ET 15-21) recognizes the region at C-terminal of ET-3 and the detection antibody specifically recognizes the region at N-terminal of ET-3.
  • Question
    Q.How should I conduct Endothelin extraction process?
  • Answer
    A.Please refer to the datasheet enclosed the product package and/or the datasheet listed on the product page.
    As several publications can be found on the page (product description), please refer to them.
    https://www.ibl-japan.co.jp/en/search/product/detail/id=3869
  • Question
    Q.Is extraction process essential for blood samples?
  • Answer
    A.Yes, it is essential. Please follow the datasheet for the extraction process using column such as SepPak.
  • Question
    Q.I have only 0.5mL blood samples. Is it possible to measure it using this ELISA?
  • Answer
    A.Yes, it is possible, however, in general speaking, the concentration of Endothelin in blood sample is low conc., the concentration could be much lower after the extraction and process. Please well consider the risk of lack of concentration prior start the measurement.
    Our recommendation is more than 2mL blood samples.
  • Question
    Q.Is composition of EIA buffer of each ELISA kit all same? Can it be mixed to use?
    ELISA common FAQ
  • Answer
    A.No it isn't. As constitute of each EIA buffer is different, it cannot be mixed with other lots or EIA buffers contained in other kind of ELISA kits.
  • Question
    Q.What is the composition of concentrated wash buffer?
    ELISA common FAQ
  • Answer
    A.It contains ordinary Tween and phosphate buffer (0.05% Tween-20 in PB).
  • Question
    Q.What is the feature of the plate?
    ELISA common FAQ
  • Answer
    A.We use plate that is flat bottom and removable strip type plate (8wellx 12 strips).
  • Question
    Q.Can I re-use standard after reconstitution?
    ELISA common FAQ
  • Answer
    A.Not recommended to re-use standard after reconstitution. Please use it at once after the reconstitution.
    Please note that there are some exceptions. One time freeze-thaw the standard is acceptable for use after reconstitution for some ELISAs.
    Please check the details on each product datasheet.
  • Question
    Q.What is different between reagent blank and test sample blank?
    ELISA common FAQ
  • Answer
    A.Reagent blank means a well is only added EIA buffer and the purpose is confirming whether the Test sample value is influenced by lack of washing process or other operations. Test sample blank means a well is added EIA buffer and HRP antibody and the purpose is to calculate the background.
  • Question
    Q.How many samples can be measured by this kit?
    ELISA common FAQ
  • Answer
    A.The pre-coated plate contained in our ELISA kit is 96 wells plate. We recommend to use 16 wells (2 slits) for standard and 80 wells (10 slits) for 40 samples in duplicate.
  • Question
    Q.What is LOD (Limit of Detection)?
    ELISA common FAQ
  • Answer
    A.It (LOD) is defined as sensitivity that is calculated using the NCCSL method. Please refer to a datasheet of each product.
  • Question
    Q.What is LOQ (Limit of Quantification)?
    ELISA common FAQ
  • Answer
    A.It (LOQ) is the lowest value of measurement (standard) range. Please refer to a datasheet of each product.
  • Question
    Q.What is the definition of Over Night (O/N) reaction?
    ELISA common FAQ
  • Answer
    A.It means that the reaction is required more than 16 hours unless otherwise specifically defined it on a datasheet of each ELISA product.
  • Question
    Q.What is the specification of quality control for ELISA product release?
    ELISA common FAQ
  • Answer
    A.The information of specification is available on individual lot specific CoA. Please contact us with your reference lot number for obtaining of specific CoA.
  • Question
    Q.What is the number (e.g. 432143214321) at the edge of strips of the plate?
    ELISA common FAQ
  • Answer
    A.According to the plate maker (ThermoFisher), it does not have any specific meaning as it is just the number of molds.
  • Question
    Q.How to wash an ELISA plate?
    ELISA common FAQ
  • Answer
    A.Washing it by an auto-washer is highly recommended.
    If it is not available, please refer to the demo video (only 2 mins) using a washing bottle.
  • Question
    Q.The wells turned black during the test with the kit.
    ELISA common FAQ
  • Answer
    A.It is possible that the wells were not washed sufficiently during the washing process after the HRP-labeled antibody reaction.
    Be sure to wash the wells enough times as described in the data sheet with washing buffer of more than 350 µL.

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