DM1 ADC ELISA 试剂盒

(1) Prior to use allow all reagents to come to room temperature. Reagents from different kit lot numbers should not be combined or interchanged.
(2) ELISA Wash Concentrate must be diluted to working solution prior to use. Please see REAGENTS section for details.
(3) Using EDI Calibrators: Reconstitute calibration stock 30702 with 0.5 mL DI-water. Dilute the reconstituted calibration stock (30702) 1:X* using the zero calibrator (30701) to obtain a level-8 standard at 1 g/mL. Further create calibrator level seven to two by 1:2 serial dilutions to obtain these calibrators with concentrations of 0.5 g/mL, 0.25 g/mL, 0.125 g/mL, 0.063 g/mL, 0.031 g/mL, 0.016 g/mL. Assay calibrators should be used within 2 hours and should be stored below -20 C. Do not exceed 3 freeze-thaw cycles.
(5) Place a sufficient number of Anti-DM1 antibody coated microwell strips in a holder to determine calibrators and unknown samples in duplicates.

(1) Add 100 µL of calibrators and test samples into the designated microwells.
(2) Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 30 minutes at 400 to 450 rpm.
(3) Immediately add 25 µL of HRP Conjugated DM1 (cat# 30683) to each well. (Note: no wash step before add the HRP Conjugated DM1)
(4) Seal the plate wells securely, cover with foil or other material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 2 hr. 10 minutes at 400 to 450 rpm.
(5) Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents. Alternatively, an automated microplate washer can be used.
(6) Add 100 µL of ELISA HRP Substrate into each of the wells.
(7) Cover the plate with aluminum foil or other material to avoid exposure to light. Incubate plate static, at room temperature for 20 minutes.
(8) Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
(9) Read the absorbance at 450 nm.

It is recommended to use a 4-parameter or log-logit calibration curve fitting.
1. Calculate the average absorbance for each pair of duplicate test results.
2. The calibration curve is generated by the corrected absorbance of all calibration levels on the ordinate against the calibrator concentration. Appropriate computer assisted data reduction programs should be used for the calculation of results. The antibody-DM1 conjugate concentrations for the test samples are read directly from the calibration curve using their respective corrected absorbance.