Guava Nexin Reagent (for early and late apoptosis detection for Cells culture)
Materials and Reagents
1. Guava Nexin Reagent (Millipore, Cat. N 4500-0450)
2. Guava ViaCount CDR (Millipore, Cat. N 4700-0050)
3. Guava System with Guava CytoSoft Software or equivalent cytometry system with ability
to detect PE and 7-AAD fluorescence.
Preparation
o Allow Guava Nexin Reagent to warm to room temperature (about 30 min)
Wash cells with 10 ml of 1X PBS
Remove PBS and place in 50 ml tubes
Dilute the ViaCount CDR 1:3 with PBS
For a T-75 cm2 flask, add 3 ml of diluted ViaCount CDR and incubate at 37C until
cells being to detach (3-5 min)
o Add 6 ml of medium with serum and pipet repeatedly to release cells from the flask
bottom
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Procedure
1. Pipette 100 l of each sample into appropriate tube/well (cell concentration between 2X104
and 1X105 cel/sample)
2. Add 100 l Guava Nexin Reagent to each tube/well
3. Incubate for 20 min at RT in the dark
4. Acquire samples on a Guava System
Updated 1/25/11 by RK