ELISA Kit for Fibroblast Growth Factor Receptor 1 (FGFR1)

CD331; FLT2, KAL2; CEK; FLG; BFGFR, N-SAM,Basic Fibroblast Growth Factor Receptor 1; Fms-Related Tyrosine Kinase-2; Pfeiffer Syndrome

Specificity

This assay has high sensitivity and excellent specificity for detection of Fibroblast Growth Factor Receptor 1 (FGFR1).
No significant cross-reactivity or interference between Fibroblast Growth Factor Receptor 1 (FGFR1) and analogues was observed.

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Fibroblast Growth Factor Receptor 1 (FGFR1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Fibroblast Growth Factor Receptor 1 (FGFR1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Stability

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.

Reagents and materials provided

ReagentsQuantityReagentsQuantity
Pre-coated, ready to use 96-well strip plate1Plate sealer for 96 wells4
Standard2Standard Diluent1×20mL
Detection Reagent A1×120µLAssay Diluent A1×12mL
Detection Reagent B1×120µLAssay Diluent B1×12mL
TMB Substrate1×9mLStop Solution1×6mL
Wash Buffer (30 × concentrate)1×20mLInstruction manual1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Fibroblast Growth Factor Receptor 1 (FGFR1). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Fibroblast Growth Factor Receptor 1 (FGFR1). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Fibroblast Growth Factor Receptor 1 (FGFR1), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Fibroblast Growth Factor Receptor 1 (FGFR1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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