Western blot analysis was performed on nuclear-enriched extracts (30 ug lysate) of RSC-96 (Lane 1), T98G (Lane 2), MCF-7 (Lane 3), A549 (Lane 4), HCT 116 (Lane 5), K-562 (Lane 6), DU 145 (Lane 7), and PC-3 (Lane 8). The blots were probed with Rabbit Anti-MeCP2 Polyclonal Antibody (Product # PA1-888, 0.5 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 ug/ml, 1:4000 dilution). A 56 kDa band corresponding to MeCP2 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C P(469) N R E E P V D S R T P V T E R V S(486)|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Western Blot (WB)||0.1-1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (IHC)||See 2 publications below|
PA1-888 detects methyl CpG binding protein 2 (MeCP2) from human, mouse and rat cells and tissues.
PA1-888 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~56 kDa protein representing MeCP2 from AtT20 cell extract.
PA1-888 immunizing peptide corresponds to amino acid residues 469-486 from human MeCP2. This sequence is completely conserved in rat and mouse MeCP2. PA1-888 immunizing peptide (Cat. # PEP-121) is available for use in neutralization and control experiments.
DNA methyltransferases methylate the 5-position of cytosine in the context of CpG dinucleotides. DNA methylation is crucial for normal embryonic development, imprinting, and X-chromosome inactivation. Methyl CpG binding proteins (MeCPs) specifically recognize methylated regions of DNA and repress transcription both directly and by association with known corepressor proteins which include members of the Sin3 and histone deactylase protein families.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Abnormalities of cell packing density and dendritic complexity in the MeCP2 A140V mouse model of Rett syndrome/X-linked mental retardation.
PA1-888 was used in immunohistochemistry to investigate the neurological abnormalities in a mouse model of Rett syndrome/X-linked mental retardation in an effort to find the corresponding mechanisms
|Jentarra GM,Olfers SL,Rice SG,Srivastava N,Homanics GE,Blue M,Naidu S,Narayanan V||BMC neuroscience (11:null)||2010|
Multiple pathways regulate MeCP2 expression in normal brain development and exhibit defects in autism-spectrum disorders.
PA1-888 was used in immunohistochemistry to investigate the regulatory mechanism for MeCP2 expression in normal brain development and in autism-spectrum disorders
|Samaco RC,Nagarajan RP,Braunschweig D,LaSalle JM||Human molecular genetics (13:629)||2004|