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Rat CRH(Corticotropin Releasing Hormone) ELISA Kit

  • Cat.No.:E-EL-R0270

  • Reactivity: Rat

To Purchase E-EL-R0270

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

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.
ItemSpecificationsStorage
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) 1.00 2.70 8.90 1.10 2.70 8.20
Standard deviation 0.10 0.10 0.40 0.10 0.10 0.40
CV (%) 10.00 3.70 4.49 9.09 3.70 4.88

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) 84-99 91
EDTA plasma(n=8) 86-98 91
Cell culture media(n=8) 89-99 94

Linearity

Samples were spiked with high concentrations of Rat CRH and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

SynonymsCRF, Corticoliberin

Assay Procedures

elisa assay procedure 1

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

elisa assay procedure 2

2. Aspirate and wash the plate for 3 times

elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

elisa assay procedure 5

5. Add 50μL Stop Solution

elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Citations

From now on, if you have published a paper by using any of our products since 1/1/2019, fill out the “Elabscience Publication Reward Application Form”carefully and send it to orders@elabscience.com, we will get back to you with the reward after we confirm it ASAP!

View more details about our Publication Reward >>

  1. Drug Metabolism and Disposition (2018) IF: 4.242
    Activation of 5-HT1A receptors in the hypothalamic paraventricular nuclei (PVN) negatively regulates cytochrome P450 expression and activity in rat liver.

    DOI: 10.1124/dmd.117.079632

    PMID: 29555828

  2. Neurogastroenterology & Motility (2019) IF: 3.803
    Metformin Prevents Colonic Barrier Dysfunction by Inhibiting Mast Cell Activation in Maternal Separation-Induced IBS-like Rats

    DOI: 10.1111/nmo.13556

    PMID: 30740845

    Sample: Plasma,Cell culture supernatant
  3. Applied Physiology, Nutrition, and Metabolism (2020) IF: 2.522
    Anxiolytic and antioxidant effects of Euterpe oleracea Mart. (A?aí) seed extract in the adult rat offspring submitted to periodic maternal separation

    DOI: 10.1139/apnm-2020-0099

    PMID: 32516542

    Sample: Plasma
  4. International Journal of Clinical And Experimental Medicine (2017) IF: 1.069
    Higher Intravenous Anesthesia Dose Protects Memory In Young Rats And Is Associated With Decreased Release Of Corticotropin-releasing Factor.
    Sample: Tissue homogenate
  5. Heliyon (2019)
    Adverse effects of noise stress on glucose homeostasis and insulin resistance in Sprague-Dawley rats

    DOI: 10.1016/j.heliyon.2019.e03004

    PMID: 31890958

    Sample: Serum
  6. NigEr?an Society for Experimental Biology (2017)
    Pregnancy and Fetal Outcomes Following Metformin Use in Diabetic Rats

    DOI: 10.1530/endoabs.50.P354

    PMID: 99999999

    Sample: Plasma
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