CLIA Kit for Adrenocorticotropic Hormone (ACTH)

Brand

Cloud-Clone

short description

Chemiluminescent immunoassay for Antigen Detection.
Research focus: Endocrinology; Hormone metabolism; Urology; ;

spezie

Rattus norvegicus (Rat)

Cross Reaction

This assay has high sensitivity and excellent specificity for detection of Adrenocorticotropic Hormone (ACTH).
No significant cross-reactivity or interference between Adrenocorticotropic Hormone (ACTH) and analogues was observed.

detection target

Adrenocorticotropic Hormone
coating antibody: Mouse

alias

Corticotropin; Adrenocorticotropin

sensitivity

The minimum detectable dose of this kit is typically less than 4.52 pg/mL

detection range

11.72-3000 pg/mL

certificate

ISO 9001:2008, ISO 13485:2003

sample type

serum, plasma and other biological fluids

pack size

48T, 96T, 5x96T, 10x96T

Reproducibility

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Adrenocorticotropic Hormone (ACTH) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Adrenocorticotropic Hormone (ACTH) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Assay time

2h

storage and Stability

All the kits should stored according to the manual.
The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree upon receipt while the others should be at 4 degree.
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.

Principle of the Elisa

Competitive Inhibition
Test principle: The microplate provided in this kit has been pre-coated with a monoclonal antibody specific to Adrenocorticotropic Hormone (ACTH). A competitive inhibition reaction is launched between biotin labeled Adrenocorticotropic Hormone (ACTH) and unlabeled Adrenocorticotropic Hormone (ACTH) (Standards or samples) with the pre-coated antibody specific to Adrenocorticotropic Hormone (ACTH). 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 Adrenocorticotropic Hormone (ACTH) in the sample. Then the mixture of substrate A and B is added to generate glow light emission kinetics. Upon plate development, the intensity of the emitted light is reverse proportional to the Adrenocorticotropic Hormone (ACTH) level in the sample or standard.

Kit Content

Pre-coated, ready to use 96-well strip plate: 1
Plate sealer for 96 wells: 4
Standard: 2
Standard Diluent: 1x20mL
Detection Reagent A: 1
Assay Diluent A: 1x12mL
Detection Reagent B: 1x120µ L
Assay Diluent B: 1x12mL
Reagent Diluent: 1x300µ L
Stop Solution: 1x6mL
TMB Substrate: 1x9mL
Wash Buffer (30 x concentrate): 1x20mL
Instruction manual:1

Assay procrdure summery

1. Prepare all reagents, samples and standards;
2. Add 50µ L standard or sample to each well.
        And then add 50µ L prepared Detection Reagent A immediately.
        Shake and mix. Incubate 1 hour at 37° C;
3. Aspirate and wash 3 times;
4. Add 100µ L prepared Detection Reagent B. Incubate 30 minutes at 37° C;
5. Aspirate and wash 5 times;
6. Add 100µ L Substrate Solution. Incubate 10 minutes at 37° C;
7. Read RLU value immediately.