p66 alpha Overexpression Lysate

Western Blot: p66 alpha Overexpression Lysate (Adult Normal) [NBP2-06953] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for p66 alpha.

• Reactivity: Hu
• Applications: WB

p66 alpha Overexpression Lysate Summary

• Description:

  p66 alpha Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC204068
  
  Accession#: NM_017660
  
  Protein Tag: C-MYC/DDK
  
  You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.
• Gene: GATAD2A

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.
  
  Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
• Theoretical MW: 67.9 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Buffer: RIPA buffer

Lysate Details for Array

• Type: Overexpression

Notes

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Alternate Names for p66 alpha Overexpression Lysate

• FLJ20085
• FLJ21017
• GATA zinc finger domain containing 2A
• GATA zinc finger domain-containing protein 2A
• hp66alpha
• p66 alpha
• p66alpha
• transcriptional repressor p66-alpha

Background

Methylation of cytosine at CpG dinucleotides is a common feature of many higher eukaryotic genomes (1-3). Increasing evidence indicates that recruitement of histone deacetylase complexes by methyl-CpG-binding proteins (MeCP) is a major mechanism of methylated DNA silencing. P66, a zinc finger-containing protein is one of the components of the MeCP1 complex (4). P66 and p68, the two components of MeCP1 are different forms of the same zinc finger-containing protein and are conserved from C.elegans to humans. Homologs of p66 from different organisms revealed two highly conserved regions, CR1 and CR2. While CR1 is involved in the association of p66 with other MeCP1 components, CR2 plays an important role in targeting p66 and MBD3 to specific loci (4).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Bioinformatics

• Gene Symbol: GATAD2A