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Mouse CRH(Corticotropin Releasing Hormone) ELISA Kit

Citations(5)Uniprot : Q8CIT0

Cat.No.:E-EL-M0351
Reactivity: Mouse

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96T
48T
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96T*5
96T*10

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Product Details

Properties

Assay type	Competitive-ELISA
Format	96T/48T
Assay time	2.0h
Detection range	0.31-20 ng/mL
Sensitivity	0.19 ng/mL
Sample type &Sample volume	serum, plasma and other biological fluids; 50μL
Specificity	This kit recognizes CRH in samples. No significant cross-reactivity or interference between CRH and analogues was observed.
Reproducibility	Both intra-CV and inter-CV are < 10%.
Application	This ELISA kit applies to the in vitro quantitative determination of CRH concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with CRH. During the reaction, CRH in the sample or standard competes with a fixed amount of CRH on the solid phase supporter for sites on the Biotinylated Detection Ab specific to 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 CRH in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.

Item	Specifications	Storage
Micro ELISA Plate(Dismountable)	96T: 8 wells ×12 strips 48T: 8 wells ×6 strips	-20℃, 6 months
Reference Standard	96T: 2 vials 48T: 1 vial
Concentrated Biotinylated Detection Ab (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×)	96T: 1 vial, 120 μL 48T: 1 vial, 60 μL	-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent	1 vial, 20 mL	2-8°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	2-8℃(Protect from light)
Stop Solution	1 vial, 10 mL	2-8℃
Plate Sealer	5 pieces
Manual	1 copy
Certificate of Analysis	1 copy

Technical Data

Precision

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

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (ng/mL)	1.10	3.20	7.00	1.20	3.50	7.60
Standard deviation	0.10	0.20	0.30	0.10	0.20	0.20
CV (%)	9.09	6.25	4.29	8.33	5.71	2.63

Recovery

The recovery of CRH 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)	94-108	100
EDTA plasma(n=8)	92-108	98
Cell culture media(n=8)	88-101	94

Target Information

Database Links	SwissProt: Q8CIT0
Synonyms	CRF, Corticoliberin
Research Area	Cancer, Neuroscience, Signal transduction

Assay Procedures

	1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
	2. Aspirate and wash the plate for 3 times
	3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
	4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
	5. Add 50μL Stop Solution
	6. Read the plate at 450nm immediately. Calculation of the results

Citations

CHINESE JOURNAL OF CHEMISTRY (2021) IF: 6
Uncarialins J‐M from Uncaria rhynchophylla and their anti‐depression mechanism in unpredictable chronic mild stress‐induced mice via activating 5‐HT1A receptor

DOI: 10.1002/cjoc.202000652

Sample: lung
PHYTOMEDICINE (2020) IF: 4.268
Uncaria rhynchophylla ameliorates unpredictable chronic mild stress-induced depression in mice via activating 5-HT1A receptor: Insights from transcriptomics

DOI: 10.1016/j.phymed.2020.153436

Sample: serum
Frontiers in Cellular and Infection Microbiology (2017) IF: 4.3
Microbiota Modulate Anxiety-Like Behavior and Endocrine Abnormalities in Hypothalamic-Pituitary-Adrenal Axis

DOI: 10.3389/fcimb.2017.00489

Sample: Tissue homogenate
Scientific Reports (2021) IF: 4.38
The role of inflammatory cytokines in anemia and gastrointestinal mucosal injury induced by foot electric stimulation

DOI: 10.1038/s41598-021-82604-7

PMID: 33542312
Sample: Serum,Tissue homogenate(hypothalamus,hypophysis )
JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY (2018) IF: 2.478
High activity of the endogenous opioid system and acute but not chronic stress influence experimental colitis development in mice

DOI: 10.26402/jpp.2018.5.11

Sample: plasma

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