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EdU Staining Using Kit Click-IT EdU HCS Assay

Author: Amit Sharma, updated date: , view: 292, Q&A: 0
Tags: Cell Culture and EdU


Day 1. 

Treat cells with 1X EdU from 1000x stock. . Add 0.2 ul/2 ml complete media.

Day 2. 

  1. Wash cells with 1x PBS

  2. Fixing: add 500 ul of 4% PFA. Incubate at RT for 15 min.

  3. Wash 1x PBS X2

  4. Add 0.3% Triton X-100, incubate at RT FOR 10 min.

  5. Wash 1x PBS X2

  6. Prepare following Rn.

  7. 1x click IT buffer:               1.7 ml
    CuSO4 (Component D):         80 ul
    Alexa Fluor (Component B)   5ul
    1x ClickIT buffer additive     200ul
    Final vol                                       2 ml 

  8. Add 250 ul/well incubate at RT 30 min.

  9. Wash cells with 100 ul of rn. rinse buffer

  10. Add DAPI

  11. Mount

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