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AICAR(AICA Riboside) ELISA Kit (UNES00064)

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SKU:
UNES00064
$567

Description

AICAR(AICA Riboside) ELISA Kit

This ELISA kit uses Competitive-ELISA as the method. The microtiter plate provided in this kit has been pre-coated with AICAR. During the reaction, AICAR in the sample or standard competes with a fixed amount of  AICAR on the solid phase supporter for sites on the Biotinylated Detection Ab specific to  AICAR. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by adding Stop Solution and the color change is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of   AICAR in the samples is then determined by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityUniversal
Detection MethodColormetric
Detection Range12.50-800 pg/mL
Sensitivity7.50 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes AICAR in samples. No significant cross-reactivity or interference between AICAR and analogues was observed.

Typical 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.

Concentration
(pg/mL)
O.D Average Corrected
8002.457
2.497
2.4772.418
4001.562
1.614
1.5881.529
2000.935
0.921
0.9280.869
1000.448
0.45
0.4490.39
500.246
0.24
0.2430.184
250.173
0.147
0.160.101
12.500.106
0.116
0.1110.052
00.051
0.067
0.059--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)42.3976.75374.4244.8175.75400.25
Standard deviation2.233.2219.512.723.3617.97
C V (%)5.264.205.216.074.444.49

Recovery

The recovery of AICAR 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=5)86-9991
EDTA plasma (n=5)86-9993
Cell culture media (n=5)87-9893

Linearity

Samples were spiked with high concentrations of AICAR 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)
1:2Range (%)92-10997-11597-112
Average (%)100105105
1:4Range (%)89-10582-9783-93
Average (%)968888
1:8Range (%)86-9883-9885-98
Average (%)929092
1:16Range (%)92-10781-9382-96
Average (%)988688

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells ×12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100×)1 vial, 120 uL
Concentrated HRP Conjugate (100×)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25×)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy
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Additional Information

Reactivity:
Universal
ELISA Type:
Sandwich
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