Catalog No: RE3022H
Human CRH(Corticotropin Releasing Hormone) ELISA Kit
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48T

$299.00

96T

$399.00

Product Details

Species:

Human

Alternative Name:

CRF, Corticoliberin

Uniport ID:

P06850

Assay Type:

Competitive

unit:

pg/mL

Standard:

250pg/mL

Sensitivity:

2.35 pg/mL

Detection Range:

3.91-250pg/mL

Assay Time:

1.5H

Sample Type:

Serum, Plasma, Tissue homogenate and Other biological samples;Sample Volume=50μL

Research:

Endocrinology;Neuro science;Hormone metabolism;

Test Principle:

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with Human CRH. During the reaction, Human CRH in the sample or standard competes with a fixed amount of Human CRH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to Human CRH. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of Human CRH in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Technical Data

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.
pg/mL OD Corrected
250 0.159
125 0.336
62.5 0.541
31.25 0.851
15.63 1.193
7.82 1.575
3.91 1.835
0 2.466

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level were tested on 3 different plates, 20 replicates in each plate, respectively.

Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean(ng/mL) 11.36 51.6 92.18 1.95 42.79 81.13
Standard deviation 0.3 2.67 5 0.82 2.77 4.2
C V (%) 2.85 5.66 3.4 3.06 5 2.6

Rate of recovery

The recovery of spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
Sample Type Range(%) Average Recovery(%)
Serum (n=8) 85-97 91
EDTA plasma (n=8) 85-97 91
Cell culture media (n=8) 83-99 95

Linear

Samples were spiked with high concentrations of target proteins 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)
1:2 Range (%) 95-103 88-104 89-97
Average (%) 102 98 94
1:4 Range (%) 80-96 88-104 85-94
Average (%) 87 100 87
1:8 Range (%) 93-106 86-102 87-101
Average (%) 99 101 98
1:16 Range (%) 87-98 80-93 82-95
Average (%) 89 89 88

Assay Procedures

Description