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Home Over-expression Lysates YY1 Lysate

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YY1 Over-expression Lysate Product

DNA-binding activity

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Specifications Citations Related Products Product Documents
Cat. # Description Price Availability
LY401166 $280 5 days
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Expression cell line:Human HEK293T cell line
Expression cDNA Clone:TrueORF Clone RC207198
Epitope tag:C-terminal Myc/DDK*
Detection Antibodies:Anti-DDK Antibody (TA50011)
Accession Number:NM_003403   NP_003394
Other Names:YY1,DELTA; INO80S; NF-E1; UCRBP; YIN-YANG-1
Bioactivity: DNA-binding activity: view data
Predicted MW:44.5 kDa
Components: 1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer
1 vial of 100 µg empty vector transfected control cell lysate in RIPA buffer
1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 100mM DTT)
Storage: The lysate is shipped with dry ice. Upon receiving, store the sample at -20ºC. Lysate samples are stable for 12 months from date of receipt when stored at -20ºC. Avoid repeated freeze-thaw cycles.
Lysate samples can be diluted with 2xSDS Sample Buffer provided. After dilution, the protein sample should be aliquoted and stored at -20ºC for long term storage. Prior to SDS-PAGE fractionation, boil the lysate for 5 minutes.
Preparation: HEK293T cells in 10-cm dishes were transiently transfected with MegaTran Transfection Reagent (TT200002) and 5ug TrueORF cDNA plasmid. 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 (Sigma), 1mM PMSF and 1mM Na3VO4), and then centrifuged to clarify the lysate. Protein concentration was measured by BCA kit (Thermo Scientific Inc.). Cell lysates were aliquoted and stored at -20ºC before shipping.
Protein Families: Transcription FactorsDruggable Genome
Western validation with an anti-DDK antibody*
L: Control HEK293 lysate
R: Over-expression lysate

 

*: DDK-tag is the same as FLAG tag. Flag® is a registered trademark of Sigma-Aldrich
**: Delivery time is an estimate in business days. Occasional delays may occur due to unforeseen complexities in the preparation of your construct. International customers may expect an additional 1-2 weeks in shipping

DNA-binding activity of YY1 was measured in OriGene over-expression lysate LY401166 and a control lysate. Three microliters of each lysate was tested with a transcription factor binding assay utilizing YY1-specific DNA sequences. The high level of activity observed in the over-expression lysate compared to the control lysate demonstrates that the expressed YY1 is biologically active in the lysate.

 

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