Recombinant MeCp2

 

Catalog # : EPX-001-RBV

 

Source : Human

 

Expressed in : SF9 cells

 

Quantity : 10g of active recombinant MeCp2 at 0.5g /l

 

 

 

Background:

 

MeCp2, methyl CpG binding protein 2, is a chromosomal protein that binds to methylated DNA (1). It can bind specifically to a single methyl-CpG pair and is not influenced by sequences flanking the methyl-CpGs (1). MeCp2 mediates transcriptional repression through interaction with histone deacetylase and the corepressor SIN3A (2). Defects in MECP2 are the cause of Rett syndrome, a progressive X-linked neurologic developmental disorder that causes mental retardation in females (3).

 

 

Protein details:


 Recombinant human N-terminal His tagged MeCp2 was produced in SF9 insect cells, purified using FPLC and formulated in a storage buffer containing 20mM Tris-Cl pH 7.6, 1mM EDTA, 0.15 M NaCl, 10% glycerol, 0.5mM PMSF and 1mM DTT. Protein concentration was determined by spectrometry. >95% purity by SDS-PAGE.

 

 

Quality control:

 

Each lot has been evaluated by 10% Tris Glycine SDS-PAGE.

 

center  

SDS-PAGE gel of recombinant MeCp2 (Lane 2).  Lane 1, protein molecular weight marker.



Double strand methylated DNA (lane 1) was incubated with increasing amount of recombinant MeCp2 protein (lane 2-4) and run on native PAGE .

 

MeCp2 Gel shift assay:

 

A 70 bp methylated DNA (50 ng) was mixed with competitor DNA (200 ng), followed by incubation with various amounts of MeCP2 in binding buffer (50 mM NaCl, 20 mM Tris-Hcl, 0.25 mM EDTA [pH 7.65]) at room temperature for 30 min. DNA was immediately electrophoresed on 6% native polyacrylamide gel 0.5 TG (0.025 M Tris, 0.192 M glycine) and run at room temperature.

 

 

Storage:

 

-80C

 

 

Guarantee:

 

For research use only. Products guaranteed stable for 2 years from date of receipt when stored properly.

 

 

Purity:

 

>98% purity by SDS PAGE.

 

 

Sequence:


MeCp2 (Mw: 52,441 Da.)
MVAGMLGLREEKSEDQDLQGLKDKPLKFKKVKKDKKEEKEGKHEPVQPSAHHSAEPAEAG
KAETSEGSGSAPAVPEASASPKQRRSIIRDRGPMYDDPTLPEGWTRKLKQRKSGRSAGKY
DVYLINPQGKAFRSKVELIAYFEKVGDTSLDPNDFDFTVTGRGSPSRREQKPPKKPKSPK
APGTGRGRGRPKGSGTTRPKAATSEGVQVKRVLEKSPGKLLVKMPFQTSPGGKAEGGGAT
TSTQVMVIKRPGRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETV
LPIKKRKTRETVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASS
PPKKEHHHHHHHSESPKAPVPLLPPLPPPPPEPESSEDPTSPPEPQDLSSSVCKEEKMPR
GGSLESDGCPKEPAKTQPAVATAATAAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTP
VTERVS



 

References:

1.    Meehan et al., Cell, 58 (1989), pp. 499–507
2.    Nan et al., Nature. 1998 May 28;393(6683):386-9.
3.    Amir et al., Nat. Genet. 23:185-188(1999).


 

 

 


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