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Mouse IFN-γ(Interferon Gamma) ELISA Kit

Elisa Kit

Mouse IFN-γ(Interferon Gamma) ELISA Kit

Catalogue No BSEK-414M
Species Mouse
Size 96 T | 48 T
Price POR
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SKU: BSEK-414M Categories: ,
Sensitivity 18.75pg/ml
Range 31.25-2000pg/ml (1ng=8430mIU)
Alternative Name Interferon gamma ELISA Kit, IFN-gamma ELISA Kit, IFNG ELISA Kit
UniProt ID P01580
Sample Type Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
Detection Method Sandwich ELISA, Double Antibody
Detection Wavelength OD450
Reaction Duration 4 hours
Storage 2-8°C(Sealed), Don't cryopreserve
Specificity Specifically binds with IFN-γ , no obvious cross reaction with other analogues.
Principle This kit is based on Double antibody-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with anti IFN-γ antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with IFN-γ bound to precoated antibody. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of IFN-γ in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.

 

 

 

This product is detected by QC department and meets performance required in the manual. (Laboratory Humidity: 20%-60%; Temperature: 18°C -25°C; Equilibrate TMB substrate to 37°C before staining. After adding into the ELISA wells, incubate for 15min at 37°C in dark.)

Due to different assay environments and operations, assay data below and standard curve are provided for reference. Experimenters should establish standard curve according to their own assay.

STD.(pg/ml) OD-1 OD-2 Average
0 0.059 0.062 0.06
31.25 0.104 0.108 0.106
62.5 0.149 0.155 0.152
125 0.217 0.226 0.222
250 0.382 0.398 0.39
500 0.646 0.673 0.66
1000 1.299 1.352 1.325
2000 2.297 2.391 2.344

Add a certain amount of IFN-γ into the sample. Calculate the recovery by comparing the measured value with the expected amount of IFN-γ in the sample.

Sample Type Recovery Range(%) Average(%)
serum(n=10) 91-100 97
EDTA plasma(n=10) 87-105 96
Heparin plasma(n=10) 85-102 92

Dilute the sample with a certain amount of IFN-γ at 1:2, 1:4 and 1:8 to get the recovery range.

Sample Type 1:2 1:4 1:8
serum(n=10) 89-99% 89-99% 84-100%
EDTA plasma(n=10) 91-100% 83-92% 80-93%
Heparin plasma(n=10) 85-105% 82-99% 82-87%

Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate.

Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates.

Item Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/ml) 62.47 235.1 1003.7 60.14 240.8 988.9
Standard deviation 3.52 12.06 51.99 3.06 12.09 53.2
CV(%) 5.63 5.13 5.18 5.09 5.02 5.38

Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data.

ELISA kit(n=5) 37°C for 1 month 2-8°C for 6 months
Average(%) 80 95-100