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p66 alpha Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-06953

Novus Biologicals, part of Bio-Techne
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NBP2-06953

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for p66 alpha Overexpression Lysate

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.

Product Specifications

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.

TMW

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.

Type

Overexpression

Scientific Data Images for p66 alpha Overexpression Lysate

Western Blot: p66 alpha Overexpression Lysate [NBP2-06953]

Western Blot: p66 alpha Overexpression Lysate [NBP2-06953]

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.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: p66 alpha

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).

Alternate Names

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

Gene Symbol

GATAD2A

Additional p66 alpha Products

Product Documents for p66 alpha Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for p66 alpha Overexpression Lysate

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.

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.

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