Human CRN (Corin) ELISA Kit (HUES01329)
- Product Type:
- ELISA Kit
- 96 Assays
- ELISA Type:
- ATC2, Lrp4, PEE5, TMPRSS10, Atrial Natriuretic Peptide-Converting Enzyme
- Tested Sample Types:
- Serum, plasma and other biological fluids
|Detection Range:||78.13-5000 pg/mL|
|Sample Volume Required Per Well:||100µL|
|Sample Type:||Serum, plasma and other biological fluids|
|Specificity:||This kit recognizes Human CRN in samples. No significant cross-reactivity or interference between Human CRN and analogues was observed.|
This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human CRN. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human CRN and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human CRN, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human CRN. The concentration of Human CRN in samples can be calculated by comparing the OD of the samples to the standard curve.
|UniProt Protein Function:||CORIN: Serine-type endopeptidase involved in atrial natriuretic peptide hormone (NPPA) processing. Converts through proteolytic cleavage the non-functional propeptide NPPA into the active hormone, thereby regulating blood pressure in heart and promoting natriuresis, diuresis and vasodilation. Proteolytic cleavage of pro-NPPA also plays a role in female pregnancy by promoting trophoblast invasion and spiral artery remodeling in uterus. Also acts as a regulator of sodium reabsorption in kidney. May also process pro-NPPB the B-type natriuretic peptide. Defects in CORIN are the cause of pre-eclampsia/eclampsia 5 (PEE5); also known as gestational proteinuric hypertension. Preeclampsia is a pregnancy-associated disease with maternal symptoms but placental origin. Unlike most other human disorders, it impacts 2 individuals, the mother and her child, both of whom can be severely affected. Belongs to the peptidase S1 family. 2 isoforms of the human protein are produced by alternative splicing.|
|UniProt Protein Details:|
Protein type:EC 3. 4. 21. -; Membrane protein, integral; Cell surface; Protease
Chromosomal Location of Human Ortholog: 4p12
Cellular Component: plasma membrane
Molecular Function:serine-type endopeptidase activity
Biological Process: female pregnancy; peptide hormone processing; regulation of blood pressure; regulation of systemic arterial blood pressure by atrial natriuretic peptide
Disease: Preeclampsia/eclampsia 5
|NCBI Summary:||This gene encodes a member of the type II transmembrane serine protease class of the trypsin superfamily. Members of this family are composed of multiple structurally distinct domains. The encoded protein converts pro-atrial natriuretic peptide to biologically active atrial natriuretic peptide, a cardiac hormone that regulates blood volume and pressure. This protein may also function as a pro-brain-type natriuretic peptide convertase. Multiple alternatively spliced transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jun 2013]|
|NCBI GenInfo Identifier:||317373348|
|NCBI Gene ID:||10699|
|NCBI Accession:||Q9Y5Q5. 2|
|UniProt Secondary Accession:||Q9Y5Q5,Q2TBD2, Q4W5E5, Q4W5G6, Q9UHY2, B0ZBE3,|
|UniProt Related Accession:||Q9Y5Q5|
|Molecular Weight:||113,297 Da|
|NCBI Full Name:||Atrial natriuretic peptide-converting enzyme|
|NCBI Synonym Full Names:||corin, serine peptidase|
|NCBI Official Symbol:||CORIN|
|NCBI Official Synonym Symbols:||CRN; ATC2; Lrp4; PEE5; TMPRSS10|
|NCBI Protein Information:||atrial natriuretic peptide-converting enzyme|
|UniProt Protein Name:||Atrial natriuretic peptide-converting enzyme|
|UniProt Synonym Protein Names:||Corin; Heart-specific serine proteinase ATC2; Pro-ANP-converting enzyme; Transmembrane protease serine 10|
|Protein Family:||Atrial natriuretic peptide-converting enzyme|
|UniProt Gene Name:||CORIN|
|UniProt Entry Name:||CORIN_HUMAN|
As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human CRN were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human CRN were tested on 3 different plates, 20 replicates in each plate.
|Intra-assay Precision||Inter-assay Precision|
|C V (%)||5.94||4.13||4.00||6.70||4.62||3.21|
The recovery of Human CRN spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
|Sample Type||Range (%)||Average Recovery (%)|
|EDTA plasma (n=5)||91-103||97|
|Cell culture media (n=5)||88-101||96|
Samples were spiked with high concentrations of Human CRN and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.
|Serum (n=5)||EDTA plasma (n=5)||Cell culture media (n=5)|
An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.
|Micro ELISA Plate(Dismountable)||8 wells ×12 strips||-20°C, 6 months|
|Reference Standard||2 vials|
|Concentrated Biotinylated Detection Ab (100×)||1 vial, 120 µL|
|Concentrated HRP Conjugate (100×)||1 vial, 120 µL||-20°C(shading light), 6 months|
|Reference Standard & Sample Diluent||1 vial, 20 mL||4°C, 6 months|
|Biotinylated Detection Ab Diluent||1 vial, 14 mL|
|HRP Conjugate Diluent||1 vial, 14 mL|
|Concentrated Wash Buffer (25×)||1 vial, 30 mL|
|Substrate Reagent||1 vial, 10 mL||4°C(shading light)|
|Stop Solution||1 vial, 10 mL||4°C|
|Plate Sealer||5 pieces|
|Product Description||1 copy|
|Certificate of Analysis||1 copy|
- Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
- Aliquot 100 µL of standard solutions into the standard wells.
- Add 100 µL of Sample / Standard dilution buffer into the control (zero) well.
- Add 100 µL of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
- Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37 °C.
- Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
- Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.