The Avelumab ELISA Kit (SKU: GXEL00253) is a quantitative competitive ELISA for the measurement of Avelumab in plasma, serum. The assay has a detection range of 62.5 - 4,000 ng/mL and a sensitivity of 18.5 ng/mL.
This assay employs the quantitative competitive enzyme immunoassay technique. Recombinant Human CD274 has been pre-coated onto a microplate. Standards or samples are premixed with biotin-labeled antibody and then pipetted into the wells. Avelumab in the sample competitively binds to the pre-coated protein with biotin-labeled Avelumab. After washing away any unbound substances, Streptavidin-HRP is added to the wells. Following a wash to remove any unbound enzyme reagent, a substrate solution is added to the wells and color develops in inversely proportion to the amount of Avelumab bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Name:
Avelumab ELISA Kit
SKU:
GXEL00253
Size:
96 Assays
Reactivity:
Human
Assay Type:
Competitive
Detection Method:
Colorimetric (450 nm)
Sensitivity:
18.5 ng/mL
Range:
62.5 - 4,000 ng/mL
Precision:
Intra-assay CV ≤ 7.5%; inter-assay CV ≤ 10.4%
Recovery:
88-89% (average, by sample type and dilution)
Sample Type:
Plasma, Serum
UniProt:
Q9NZQ7
Storage:
2-8°C for 12 months.
Stability:
When the kit was stored at the recommended temperature for 12 months, the signal intensity decreased by less than 30%. For unopened kits, if you want to prolong the storage time, please store the Standard, Detection A, Detection B and Microplate at - 20 °C, the rest reagents should be store at 4°C.
Note:
For Research Use Only. Not for diagnostic procedures.
Precision:
Intra 1
Intra 2
Intra 3
Inter 1
Inter 2
Inter 3
n
16
16
16
24
24
24
Mean (ng/mL)
1724.8
427.8
100.8
2011.1
499.0
112.5
Standard deviation
129.8
26.1
5.1
162.1
30.0
11.8
CV (%)
7.5
6.1
5.1
8.1
6.0
10.4
Recovery:
Sample
Dilution
Average (%)
Range (%)
Human serum (n=3)
1:10
88
81-105
Human serum (n=3)
1:20
89
81-95
Linearity:
Sample
Dilution
Average (%)
Range (%)
Human serum (n=3)
1:2
100
95-104
Human serum (n=3)
1:4
105
103-108
Human serum (n=3)
1:8
103
89-115
Human serum (n=3)
1:16
99
86-110
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Step
Procedure
1
Preparation: Bring all reagents and samples to room temperature and prepare reagents, working standards and samples as directed. Assay all standards and samples in duplicate.
2
Standard & Sample Incubation: Add 100 µL premixed solution (50 µL diluted standard/sample + 50 µL Detection A working solution) to each well; incubate for 1 hour at 37°C. Wash 3 times with Wash Buffer (300 µL).
3
Detection: Add 100 µL of Detection B (working solution) to each well; incubate for 30 minutes at 37°C. Wash 5 times with Wash Buffer (300 µL).
4
Color Development: Add 100 µL of Color Reagent to each well; incubate for 15 minutes at 37°C; protect from light.
5
Stop & Read: Add 50 µL of Stop Solution to each well, then read the optical density at 450 nm within 10 minutes (correction 630 nm or 620 nm).
Sample Type
Protocol
Serum
Use a serum separator tube (SST) and allow samples to clot for 30 minutes at room temperature before centrifugation for 15 minutes at 1000 x g. Remove serum and assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Plasma
Collect plasma using EDTA, heparin, or citrate as an anticoagulant. Centrifuge for 15 minutes at 1000 x g within 30 minutes of collection. Assay immediately or aliquot and store samples at ≤ -20°C. Avoid repeated freeze-thaw cycles.
Other Sample Types
For more information about how to process other sample types, (e.g., body fluids, breast milk & more), please contact our Tech Support Team at techsupport@assaygenie.com.
