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5-Hydroxyeicosatetraenoic Acid (5-HETE), General, ELISA Kit

Cloud-Clone ELISA Kits from ARP American Research Products, Inc.

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Product Code: CED739GE


Product Details Protocol

General 5-HETE ELISA Kit :: 5-Hydroxyeicosatetraenoic Acid ELISA Kit (CED739Ge)

General 5-Hydroxyeicosatetraenoic Acid (5-HETE) ELISA Kit

Product Code:CED739Ge

5-Hydroxyeicosatetraenoic Acid ELISA Kit

Method:Competitive ELISA
Sensitivity:The minimum detectable dose of this kit is typically less than 3.95ng/mL
Detection Range:9.88-800ng/mL.
The standard curve concentrations used for the ELISA's were 800ng/mL, 266.67ng/mL, 88.89ng/mL, 29.63ng/mL, 9.88ng/mL
Sample Types:Serum, plasma and other biological fluids
Specificity:This assay has high sensitivity and excellent specificity for detection of 5-Hydroxyeicosatetraenoic Acid (5-HETE).
No significant cross-reactivity or interference between 5-Hydroxyeicosatetraenoic Acid (5-HETE) and analogues was observed.
Precision:Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level 5-Hydroxyeicosatetraenoic Acid (5-HETE) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level 5-Hydroxyeicosatetraenoic Acid (5-HETE) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Stability:The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Procedure Summary:1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37oC;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50µL Stop Solution. Read at 450 nm immediately.
Test Principle:This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to 12-Hydroxyeicosatetraenoic Acid (12-HETE) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled 12-Hydroxyeicosatetraenoic Acid (12-HETE) and unlabeled 12-Hydroxyeicosatetraenoic Acid (12-HETE) (Standards or samples) with the pre-coated antibody specific to 12-Hydroxyeicosatetraenoic Acid (12-HETE). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of 12-Hydroxyeicosatetraenoic Acid (12-HETE) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of 12-Hydroxyeicosatetraenoic Acid (12-HETE) in the sample.
Assay Length:2.5 hours
Intended Use:For Research Use Only
This product is sold for laboratory research use or further manufacturing only and should not be used for human therapeutic or diagnostic applications. The information presented is believed to be accurate; however, said information and products are offered without warranty or guarantee since the ultimate conditions of use and the variability of the materials treated are beyond our control. Nothing disclosed herein is to be construed as a recommendation to use our products in violation of any patents. Under no circumstances shall ARP American Research Products, Inc. be liable for damages, whether consequential, compensatory, incidental or special, strict liability or negligence, breach of warranty or any other theory arising out of the use of the products available from ARP American Research Products, Inc. Nothing contained herein warrants that the use of the products will not infringe on the claims of any patents covering the product itself or the use thereof in combination with other products or in the operation of any process.

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