The cookie settings on this website are set to 'allow all cookies' to give you the very best experience. Please click Accept Cookies to continue to use the site.

Horse ACTH(adrenocorticotropic hormone) ELISA Kit

(No reviews yet) Write a Review
SKU:
HRFI0010
$749

Description

Product name

Horse ACTH(adrenocorticotropic hormone) ELISA Kit

Catalogue No.

HRFI0010

Alias

ACTH ELISA Kit

Detection method

Competitive ELISA, Coated with Antigen

Size

96T

Range

15.625-1000pg/ml

Sensitivity

< 9.375pg/ml

Storage

4'C for 6 months

Recovery

Matrices listed below were spiked with certain level of and the recovery rates were calculated by comparing the measured value to the expected amount of in samples.

Matrix Recovery range(%) Average(%)
serum(n=5) 88-103 93
EDTA plasma(n=5) 86-101 95
heparin plasma(n=5) 86-98 92
Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-101% 89-99% 91-105% 85-98%
EDTA plasma(n=5) 88-101% 82-100% 86-101% 82-99%
heparin plasma(n=5) 87-97% 85-100% 83-88% 90-100%
CV(%)

Intra-Assay: CV<8%
Inter-Assay: CV<10%

Note

For Research Use Only

Datasheets

ELISA Kit Technical ManualELISA Kit Technical ManualMSDS

Protocol

Competitive ELISA Protocol

The below protocol is a sample protocol for a competitive ELISA kits. Competitive ELISA kits allow for the detection and quantification of an analyte in a sample. This allows the researcher to calculate the amount of analyte present in their sample. This can be very useful when looking for increases in protein concentration, phosphorylation of proteins or decreases on protein concentrations.Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37 °C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.

Competitive ELISA Protocol

ELISA Kit Technical Manual

Procedure:

1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Add Sample and Biotin-detection antibody: Add 50µL of Standard, Blank, or Sample per well. The blank well is added with Sample / Standard dilution buffer. Immediately add 50 µL of Biotin-detection antibody working solution to each well. Cover with the Plate sealer we provided. Gently tap the plate to ensure thorough mixing. Incubate for 45minutes at 37°C. (Solutions are added to the bottom of micro ELISA plate well, avoid inside wall touching and foaming to the best of your ability.)
3.Wash: Aspirate each well and wash, repeating the process three times Wash by filling each well with Wash Buffer (approximately 350µL) using a squirt bottle, multi-channel pipette, manifold dispenser or automated washer. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and pat it against thick clean absorbent paper.
4.HRP-Streptavidin Conjugate(SABC): Add 100µL of SABC working solution to each well. Cover with a new Plate sealer. Incubate for 30minutes at 37°C.
5.Wash: Repeat the aspiration/wash process for five times.
6.TMB Substrate: Add 90µL of TMB Substrate to each well. Cover with a new Plate sealer. Incubate for about 15-20 minutes at 37°C. Protect from light. The reaction time can be shortened or extended according to the actual color change, but not more than 30minutes. When apparent gradient appeared in standard wells, you can terminate the reaction.
7.Stop: Add 50µL of Stop Solution to each well. Color turn to yellow immediately. The adding order of stop solution should be as the same as the substrate solution.
8.OD Measurement: Determine the optical density (OD Value) of each well at once, using a microplate reader set to 450 nm. You should open the microplate reader ahead, preheat the instrument, and set the testing parameters.
View AllClose

Additional Information

Reactivity:
Horse
ELISA Type:
Competitive
View AllClose