RIPAB+ EZH2 - RIPAB AND PRIMER SET

General description

GST fusion protein corresponding to amino acids 353-451 of human EZH2.

Application

RNA Binding Protein Immunoprecipitation:
RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG or Anti-EZH2 antibody and the TM=["MAGNA RIP"] RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of EZH2-associated RNA was verified by qPCR using NEAT1 Positive Control Primers.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa S3 cells (4 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG or Anti-EZH2, clone AC22 antibody and the TM=["MAGNA CHIP"] G Kit (Cat. # 17-611).

Immunoprecipitation Analysis: HeLa nuclear cell extract was Immunoprecipitated with either anti-EZH2, clone AC22 or mouse IgG and then resolved by electrophoresis, transferred to PVDF and probed with Cat. # 05-1319 anti-EZH2, clone BD43 (1:1000 dilution).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. # AP124P) and a chemiluminescence detection system.

Components

Anti-EZH2, clone AC22: Part No. CS203195 Normal Mouse IgG: Part No. CS200621 NEAT1 Positive Control Primers: Part No. CS216139

Quality

RNA Binding Protein Immunoprecipitation: RIP Lysate prepared from HeLa cells (2 X 10E7 cell equivalents per IP) were subjected to immunoprecipitation using 5 µg of either a normal mouse IgG , or Anti-EZH2 antibody and the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Cat. # 17-700). Successful immunoprecipitation of EZH2-associated RNA was verified by qPCR using NEAT1 Positive Control Primers. Please refer to the Magna RIP (Cat. # 17-700) or EZ-Magna RIP (Cat. # 17-701) protocol for experimental details.

Physical properties

~85 kDa

Physical form

Anti-EZH2, clone AC22 (mouse monoclonal IgG2a): Part No. CS203195. One vial containing 50 μg of protein G purified antibody in 50 μL 0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH7.4 before the addition of glycerol to 30%. Store at -20°C.
Normal Mouse IgG: Part No. CS200621. One vial containing 125 µg purified mouse IgG in 125 µL storage buffer containing 0.1% sodium azide. Store at -20°C.
NEAT1 Positive Control Primers: Part No. CS216139. One vial containing 75 μL of 5 μM of each primer specific for the human lncRNA NEAT1. Store at -20°C.
FOR: CTT CCT CCC TTT AAC TTA TCC ATT CAC
REV: CTC TTC CTC CAC CAT TAC CAA CAA TAC

Storage and Stability

Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.