Western blot analysis was performed on nuclear enriched extracts (30 ug lysate) of U-937 (Lane 1).The blots were probed with MBD1 Rabbit Polyclonal Antibody (Product # PA1-889, 2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). A 60 kDa band corresponding to MBD1 was observed in the cell line tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C R(593) S K D L K K P G A R K Q(605)|
|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 *|
|Western Blot (WB)||1-3 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.
PA1-889 detects human methyl CpG binding domain protein 1 (MBD1) recombinant protein.
PA1-889 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~83 kDa protein representing recombinant human MBD1.
PA1-889 immunizing peptide corresponds to amino acid residues 593-605 from human MBD1 protein. PA1-889 immunizing peptide (Cat. # PEP-130) is available for use in neutralization and control experiments.
Methyl CpG binding domain proteins (MBDs) are a family of mammalian proteins that specifically bind methylated regions of DNA. There are two methyl CpG binding complexes, MeCP1 and MeCP2. MeCP2 is comprised of a single protein also denoted MeCP2. MeCP1 is composed of four different proteins termed MBD1 (also known as PCM1), MBD2, MBD3, and MBD4 (also known as MED1). MBD1 binds to and represses transcription of symmetrically methylated CpG sequences. MBD1 is the only protein in the MeCP1 complex than contains a CXXC motif (Dnmt DNA binding motif). MBD1 also contains both a methyl CpG binding domain and a transcriptional repression domain at the C-terminus. It has been suggested that the mechanism for methylation-dependent repression is likely to involve other chromatin modifications including histone deacetylation since inhibitors of deacetylase activity have been shown to overcome MBD1-mediated gene repression.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.