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Company Source SC Mouse SC SC SC BD SC Bethyl Mouse Goat Mouse Mouse Mouse
P Ab Cell GαM NC CD 90 DLK-1 CD-133 CK-8 G1/G2 NC DαG NC h-alb
Conjugation Type FITC ----------------? FITC ----Alexa 647 FITC FITC C UC UC UC UC C C UC C C C
S Ab N/A GαM FITC GαM FITC GαM FITC GαM FITC N/A N/A DαG FITC N/A N/A N/A
Saponin Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
10 BD CD-271 11 CD31/EpCAM 12 CD34/CD45 Protocol 1. Saponin treatment 2. Unconjugated 3. Conjugated 1. Saponin a. Cells on Ice b. add 1 ml saponin c. 1500 rpm immediately for 5 minutes d. Put back on ice e. Washing with saponin only
2. Unconjuated a. Add 5ul Primary ab b. rotate manually whole set of tubes for mixing c. incubate in 37⁰C/RT for 30 minutes d. add unsterile PBS 1 ml e. 1500 rpm for 5 minutes f. add 5ul of secondary ab g. rotate manually whole set of tubes for mixing h. seal with aluminum foil—optional i. put in +4⁰C 25 minutes incubation j. read 3. Conjugated a. Add 5ul Primary ab (conj) b. rotate manually whole set of tubes for mixing c. seal with aluminum foil—optional d. put in +4⁰C 25 minutes incubation e. read