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 Home Over-expression Lysates TP53 Lysate

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TP53 Over-expression Lysate Product

TP53 Activity Verified in a DNA-binding Assay

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Specifications Related Products WB Protocol FAQs
Cat. # Description Price Availability
LY400186 $280 5 days
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Also for TP53 (NM_000546)
cDNA Clone shRNA/siRNA Primer Pair Protein Antibody

Expression cell line: Human HEK293T cell line
Expression cDNA Clone: TrueORF Clone RC200003
Epitope tag:C-terminal Myc/DDK*
Detection Antibodies:Anti-DDK Antibody (TA50011)
Application: Western Blot experiments.
Left-Control; Right -Over-expression Lysate
Accession Number:NM_000546   NP_000537
Other Names:TP53,BCC7; LFS1; P53; TRP53
Bioactivity: TP53 Activity Verified in a DNA-binding Assay: view data
Predicted MW:43.5 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 0.5 ml 2xSDS Sample Buffer (4%SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromphenol blue, 5% b-mercaptoethanol)
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: Stem cell - PluripotencyTranscription FactorsDruggable Genome
Protein Pathways: MAPK signaling pathwayCell cyclep53 signaling pathwayApoptosisWnt signaling pathwayNeurotrophin signaling pathwayMore Pathways >>

 

*: DDK-tag is the same as FLAG tag. Flag® is a registered trademark of Sigma-Aldrich

TP53 activity in the over-expression lysate (LY400186) was measured in a colorimetric DNA-binding assay. Double-stranded oligonucleotide containing the p53 consensus DNA-binding sequence was incubated with dilutions of the over-expression lysate and TP53 bound to the oligo was captured onto the surface of a microtiter plate. After washing, bound TP53 was detected with an anti-p53 primary antibody followed by an HRP-labeled secondary antibody. After initial color development, the reaction was quenched and the color intensity was measured at 450nm.

 

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