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 FOS antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with FOS 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 FOS in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E002
Lyophilized Standard
1vial
2vial
Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 month at -20°C
E003
Biotin-labeled Antibody(Concentrated, 100X)
60ul
120ul
2-8°C (Avoid Direct Light)
E034
HRP-Streptavidin Conjugate(SABC, 100X)
60ul
120ul
E024
TMB Substrate
5ml
10ml
E039
Sample Dilution Buffer
10ml
20ml
2-8°C
E040
Antibody Dilution Buffer
5ml
10ml
E049
SABC Dilution Buffer
5ml
10ml
E026
Stop Solution
5ml
10ml
E038
Wash Buffer(Concentrated, 25X)
15ml
30ml
E006
Plate Sealer
3 pieces
5 pieces
E007
Product Description
1 copy
1 copy
Recovery
Add a certain amount of FOS into the sample. Calculate the recovery by comparing the measured value with the expected amount of FOS in the sample.
Sample Type
Recovery Range(%)
Average(%)
Cell lysate(n=10)
90-105
95
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of FOS and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample
1:2
1:4
1:8
serum(n=5)
89-105%
86-95%
86-102%
EDTA plasma(n=5)
88-100%
82-101%
83-99%
heparin plasma(n=5)
85-100%
94-100%
86-100%
Sensitivity
< 0.188ng/ml
Standard Curve
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.(ng/ml)
OD-1
OD-2
Average
0
0.082
0.087
0.084
0.313
0.261
0.274
0.266
0.625
0.385
0.405
0.393
1.25
0.586
0.616
0.598
2.5
0.801
0.842
0.817
5
1.21
1.272
1.235
10
1.716
1.804
1.751
20
2.135
2.244
2.179
Assay Procedure Summary
Step 1:Add 100ul standard or sample into each well, seal the plate and statically incubate for 90 minutes at 37°C.
Washing:Wash the plate twice without immersing.
Step 2:Add 100ul biotin-antibody working solution, seal the plate and statically incubate for 60 minutes at 37°C.
Washing:Wash the plate three times and immerse for 1min each time.
Step 3:Add 100ul HRP-Streptavidin Conjugate (SABC) working solution, seal the plate and statically incubate for 30 minutes at 37°C.
Washing:Wash the plate five times and immerse for 1min each time.
Step 4:Add 90ul TMB substrate solution, seal the plate and statically incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 5:Add 50ul stop solution. Read at 450nm immediately and calculate.
Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips(Calibration is required before use.)
Sterile tubes and Eppendorf tubes with disposable tips
Absorbent paper and loading slot
Deionized or distilled water
UniProt ID
P01100
Antigen
FOS
Applications
ELISA
Applications Notes
FOS ELISA Kit allows for the in vitro quantitative determination of FOS concentrations in serum, plasma, tissue homogenates and other biological fluids.
Precision
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.