ChIPAb+ MeCP2 - ChIP Validated Antibody and Primer Set

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ MECP2 set includes a MECP2 antibody, a Normal Chicken IgY (both with bridging antibody included), and positive control primers which amplify 144 bp of a human SNRPN intron region. The MECP2 antibody and negative control IgG are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of MECP2 associated chromatin.

MeCp2 (methyl-CpG-binding protein 2) binds methylated CpG sequences in double-stranded DNA resulting in the suppression of transcription. Although MeCp2 is not essential in stem cells, it is critical for embryonic development. This protein seems to be particularly important for mature nerve cells, where it is present in high levels. The MeCp2 protein is likely to be involved in turning off ("repressing" or "silencing") several other genes. Consistent with this role, mutations of MeCp2 are linked to Rett syndrome (RTT), a progressive neurological disorder that is one of the most common causes of mental retardation in females.

~75 kDa observed

KLH-conjugated linear peptide corresponding to human MeCp2.

Chromatin Immunoprecipitation:
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from SH-SY5Ycells treated with 16 nM PMA for 72 hours (1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 8 µg of either Normal Chicken IgY with bridging antibody (Part No. CS207346), or 8 µg Anti-MeCP2 with bridging antibody (Part No. CS207345) and the Magna ChIP HiSens (Cat. # 17-10460). Successful immunoprecipitation of MeCP2 associated DNA fragments was verified by qPCR using ChIP Primers, human SNRPN (Part No. CS207347) as a positive locus, and GAPDH promoter primers (Part #22-004) as a negative locus. Please refer to the Magna ChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens(Cat. # 17-10461) protocol for experimental details. (Cat. # 17-10460) or EZ-MagnaChIP HiSens(Cat. # 17-10461) protocol for experimental details.


Chromatin Immunoprecipitation Analysis:
A representative lot of this antibody was used by an independent laboratory in ChIP. (Thatcher, K., et al. (2005). Hum. Mol. Genet. 14(6): 785-797.)

Immunoprecipitation Analysis:
A representative lot was used by an independent laboratory in IP. (Thatcher, K., et al. (2005). Hum. Mol. Genet. 14(6): 785-797.)

Western Blotting Analysis:
1 µg/mL of this antibody (prior to blending with rabbit anti-chicken bridging antibody) detected MeCp2 on 10 µg of SH-SY5Y nuclear extract.

Important Note: For western blot application of the blended antibody, use 1:680 dilution in combination with anti-rabbit HRP secondary.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

25 assays per set. Recommended use: 8 μg of blended antibody per chromatin immunoprecipitation (dependent upon biological context).

Antigen Affinity Purified

Anti-MeCP2/Rabbit Blend (Chicken & Rabbit Polyclonal), Part No. CS207345. One vial containing 200 µg purified chicken antibody blended with rabbit anti-chicken IgG bridging antibody in 105 μL storage buffer with glycerol to 38% (1.99 mg/ml final, use 4.0 µL per ChIP). Store at -20°C.

Normal Chicken IgY/Rabbit Blend, Part No. CS207346. One vial containing 200 μg of purified chicken IgY blended with purified rabbit anti-chicken IgG in 145 μL storage buffer containing 0.05% sodium azide and 30% glycerol (1.38 mg/ml final, use 5.8 µL per ChIP) . Store at -20°C.

ChIP Primers, SNRPN. Part No. CS207347. One vial containing 75 μL of 5 μM of each primer specific for a human SNRPN intron region (chr15:25200483-25200626, hg19 build). Store at -20°C.
FOR: TAC CTC CGC CTA AAA TCC CTA TG
REV: GGA AGG GCG GTG GTG ACT

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

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from SH-SY5Y cells treated with 16 nM PMA for 72 hours (1e6 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 8 µg of either Normal Chicken IgY with bridging antibody (Part No. CS207346), or 8 µg Anti-MeCP2 with bridging antibody (Part No. CS207345) and the Magna ChIP^® HiSens Kit (Cat. # 17-10460). Successful immunoprecipitation of MeCP2 associated DNA fragments was verified by qPCR using ChIP Primers, human SNRPN (Part No. CS207347).
Please refer to the MagnaChIP HiSens (Cat. # 17-10460) or EZ-MagnaChIP HiSens (Cat. # 17-10461) protocol for experimental details.

Control
Includes normal chicken IgY/rabbit IgG and primers specific for a human SNRPN intron region.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

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.