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03-900 RIPAb+™ EZH2, clone AC22- RIP Validated Antibody and Primer Set

This RIPAb+™ EZH2, clone AC22 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
Research Category: Epigenetics & Nuclear Function
03-900
1 Kit  
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Overview

Replacement Information

Key Spec Table

Species Reactivity
H, M, R, Vrt
Description
Catalogue Number03-900
DescriptionRIPAb+™ EZH2, clone AC22- RIP Validated Antibody and Primer Set
Background InformationEZH2 is a polycomb group (PcG) protein that is a catalytic subunit of the PRC2/EED-EZH2 complex, which methylates 'Lys-9' (H3K9me) and 'Lys-27' (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate 'Lys-27' of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4 and the nuclear receptor RORA. Regulates the circadian clock via histone methylation at the promoter of the circadian genes. Essential for the CRY1/2-mediated repression of the transcriptional activation of PER1/2 by the CLOCK-ARNTL/BMAL1 heterodimer; involved in the di and trimethylation of 'Lys-27' of histone H3 on PER1/2 promoters which is necessary for the CRY1/2 proteins to inhibit transcription.
References
Product Information
Components
  • Anti-EZH2, clone AC22: Part No. CS203195
    Normal Mouse IgG: Part No. CS200621
    NEAT1 Positive Control Primers: Part No. CS216139
FormatPurified
PresentationAnti-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
Quality LevelMQ200
Applications
ApplicationThis RIPAb+™ EZH2, clone AC22 -RIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Application NotesRNA 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.

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 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.
Biological Information
ImmunogenGST fusion protein corresponding to amino acids 353-451 of human EZH2.
CloneAC22
ConcentrationPlease refer to product datasheet.
HostMouse
SpecificityRecognizes EZH2, MW: ~85-100 kDa.
IsotypeIgG2a
Species Reactivity
  • Human
  • Mouse
  • Rat
  • Vertebrates
Antibody TypeMonoclonal Antibody
Gene Symbol
  • EZH2, KMT6
Purification MethodProtein G purified
UniProt Number
Molecular Weight~85 kDa
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality AssuranceRNA 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.
Usage Statement
  • 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.
Storage and Shipping Information
Storage ConditionsStable 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.
Packaging Information
Material Size1 Kit
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
03-900 04065265138697

Documentation

RIPAb+™ EZH2, clone AC22- RIP Validated Antibody and Primer Set SDS

Title

Safety Data Sheet (SDS) 

RIPAb+™ EZH2, clone AC22- RIP Validated Antibody and Primer Set Certificates of Analysis

TitleLot Number
RIPAb+ EZH2 - D2011015 D2011015