Cell Signaling Technology
Ezh2 (D2C9) XP® Rabbit mAb
#5246
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We are performing experiments to develop new proteomics screening techniques using CRISPR. This Cell Signaling Rabbit Ezh2 antibody detects small amounts of endogenous Ezh2 with great ease at the recommended (1:1000) dilution.
Western Blot
Cell lysates from different mouse and human cell lines
1:1000 Dilution in BSA, overnight incubation in cold room
5% non-fat milk in TBS-T
Anti-Rabbit-HRP, 1:3000, 1hour incubation at Room Temperature
NA
Bio-Rad Clarity HRP substrate
Protein extract from cells was prepared in RIPA lysis buffer supplied with PMSF, protease inhibitor mixture, and sodium orthovanadate. Lysates were quantified using a BCA assay kit (Thermos). 20 micrograms of proteins were subjected to SDS-PAGE and transferred onto PVDF membranes and blocked with 5% non-fat milk in TBS-T (20 mMTris, 500 mMNaCl, and 0.1% Tween 20) at room temperature for 1 h with rocking. The membranes were probed with primary Antibody (1/1000 dilution) overnight at 4°C. After washing with TBS-T, the membranes were incubated with HRP-conjugated secondary Antibodies in 5% non-fat milk/TBS-T at room temperature for 1 h. The protein–Antibody complex was detected by Bio-Rad Clarity ECL substrate. The antibody detected endogenous levels of Ezh2 in a variety of cell types, but detection in human cells was better than in mouse cells.
None
Works great in many cell types
I would use this antibody for detection of low endogenous levels of Ezh2 in a variety of cell types