This kit is based on Double antigen-Sandwich ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with antigen. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antigen. Then, it binds with Anti-Enfortumab vedotin-ejfv Ab bound to precoated antigen. 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 Anti-Enfortumab vedotin-ejfv Ab in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
- Product Name
- Anti-Enfortumab vedotin-ejfv antibody ELISA Kit
- Alias
- Anti-Enfortumab vedotin-ejfv Ab ELISA Kit
- Catalogue No.
- EU6695
- Size
- 96T
- Species
- Universal
- UniProt ID
- Q96NY8
- Sample Type
- Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples
- Detection Method
- Sandwich ELISA, Double antigen
- Detection Wavelength
- OD450
- Reaction Duration
- 4 hours
- Range
- 1.563-100ng/ml
- Sensitivity
- 0.938ng/ml
- Storage
- 2-8°C(Sealed), Don't cryopreserve.
- Specificity
- Specifically binds with Anti-Enfortumab vedotin-ejfv Ab , no obvious cross reaction with other analogues.
- Required Instruments and Reagents
-
- 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 (0.5-10μL, 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
- 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-antigen 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.
- 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 0.067 0.07 0.068 1.563 0.12 0.126 0.123 3.125 0.246 0.259 0.252 6.25 0.39 0.41 0.4 12.5 0.644 0.677 0.66 25 1.005 1.056 1.03 50 1.468 1.543 1.505 100 2.117 2.225 2.171 
- Recovery
-
Add a certain amount of Anti-Enfortumab vedotin-ejfv Ab into the sample. Calculate the recovery by comparing the measured value with the expected amount of Anti-Enfortumab vedotin-ejfv Ab in the sample.
Sample Type Recovery Range(%) Average(%) serum(n=10) 86-104 96 EDTA plasma(n=10) 86-102 91 Heparin plasma(n=10) 94-99 96 - Linearity
-
Dilute the sample with a certain amount of Anti-Enfortumab vedotin-ejfv Ab at 1:2, 1:4 and 1:8 to get the recovery range.
Sample Type 1:2 1:4 1:8 serum(n=10) 91-104% 82-99% 82-98% EDTA plasma(n=10) 86-99% 86-98% 91-100% Heparin plasma(n=10) 85-101% 83-97% 88-95% - 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 (ng/ml) 3.07 13.65 49.21 3.14 13.23 50.52 Standard deviation 0.14 0.78 2.81 0.15 0.69 3.34 CV(%) 4.68 5.68 5.72 4.68 5.22 6.61 - 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 2-8°C for 12 months Average(%) 80 95-100 85-98