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 IL-17 antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with IL-17 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 IL-17 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 IL-17 into the sample. Calculate the recovery by comparing the measured value with the expected amount of IL-17 in the sample.
Sample Type
Recovery Range(%)
Average(%)
serum(n=10)
90-102
97
EDTA plasma(n=10)
90-103
96
Heparin plasma(n=10)
90-104
98
Linearity
Dilute the sample with a certain amount of IL-17 at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type
1:2
1:4
1:8
serum(n=10)
88-98%
90-103%
85-104%
EDTA plasma(n=10)
82-99%
82-99%
84-97%
Heparin plasma(n=10)
89-98%
81-100%
92-100%
Specificity
Specifically binds with IL-17 , no obvious cross reaction with other analogues.
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.(pg/ml)
OD-1
OD-2
Average
0
0.142
0.146
0.144
6.25
0.191
0.197
0.194
12.5
0.357
0.367
0.362
25
0.464
0.478
0.471
50
0.844
0.868
0.856
100
1.279
1.317
1.298
200
1.745
1.795
1.77
400
2.138
2.2
2.169
Assay Procedure Summary
Step 1: Take out the required plate wells, add 50ul Cap/Det Ab into each well, then add 50ul Standard or Sample into individual well. (When adding standard or sample, the disposable tip lightly touches the liquid level. Change the disposable tips for different samples and standards.) Gently tap the plate for 10s to ensure thorough mixing then static incubate for 60 minutes at 37°C.
Washing: Wash the plate twice without immersion.
Step 2: Add 100ul HRP-Streptavidin (orange) into each well, seal the plate and static incubate for 30 minutes at 37°C.
Washing: Wash the plate five times without immersion.
Step 3: Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. (Accurate TMB visualization control is required.)
Step 4: Add 50ul stop solution. Read at 450nm immediately and calculate.
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.
Item
Intra-assay Precision
Inter-assay Precision
Sample
1
2
3
1
2
3
n
20
20
20
20
20
20
Mean (pg/ml)
12.66
48.71
204.7
12.74
50.17
200.3
Standard deviation
0.6
2.31
9.52
0.59
2.28
9.37
CV(%)
4.72
4.75
4.65
4.65
4.55
4.68
Stability
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
Notes
For Research Use Only
Usage Notes
Microplate reader (wavelength: 450nm) 37°C incubator (CO2 incubator for cell culture is not recommenced.) Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose) Precision single (0.5-10uL, 5-50uL, 20-200uL, 200-1000uL) 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
HS-IL-17 Accquant ELISA Kit allows for the in vitro quantitative determination of IL-17 concentrations in serum, plasma, tissue homogenates and other biological fluids.
Storage
2-8°C(Sealed), Don't cryopreserve.
Reviews of Human HS-IL-17 (High sensitive Interleukin 17) Accquant ELISA Kit