Intact MMAE ADC ELISA

Additional Information

Assay Procedure

1. Bring all reagents to room temperature (18-25°C) before use. Mix gently without causing foam.
2. Add 25 µL of calibrators and diluted 1:100 test samples into the designated microwells. Tap the plate gently.
3. Immediately add 100 µL of Assay Buffer
4. Seal the plate wells securely, cover with foil or similar material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 1 hour 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.
6. Alternatively, an automated microplate washer can be used.
7. Add 100 µL of MMAE Tracer Antibody to each well. Tap the plate gently.
8. Seal the plate wells securely, cover with foil or similar material to protect from light, and rotate on an ELISA plate shaker (small orbit radius) for 30 minutes at 400 to 450 rpm.
9. Wash each well 5 times by dispensing 350 µL of working wash solution into each well and then completely aspirating the contents.
10. Alternatively, an automated microplate washer can be used.
11. Add 100 µL of ELISA HRP Substrate into each of the wells.
12. Cover the plate with aluminum foil or similar material to avoid exposure to light. Incubate plate static, at room temperature for 20 minutes.
13. Immediately add 100 µL of ELISA Stop Solution into each of the wells. Mix gently.
14. Read the absorbance at 450 nm with reference filter at 620 nm.

Typical Standard Curve

Specificity

This Eagle Biosciences Intact MMAE ADC ELISA Assay Kit doesn’t show any cross reactivity to DM1-ADC. This assay does cross react to MMAF-ADC.   The Eagle Biosciences Intact MMAE ADC ELISA Assay Kit is a highly sensitive “sandwich” test kit is intended for use in the quantitative determination of antibody MMAE conjugate level in serum or plasma of human, mouse, rat, primate, etc.