Home Over-expression Lysates GATA4 Lysate|
GATA4 Over-expression Lysate Product
Also for GATA4 (NM_002052)
|Expression cell line:
||Human HEK293T cell line
|Expression cDNA Clone:
||TrueORF Clone RC210945|
|Epitope tag:||C-terminal Myc/DDK*|
|Detection Antibodies:||Anti-DDK Antibody (TA50011)|
||Western Blot experiments.
Left-Control; Right -Over-expression Lysate
|Accession Number:||NM_002052 NP_002043|
|Other Names:||GATA4,ASD2; VSD1
|Predicted MW:||44.4 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.
||ES Cell Differentiation/IPSEmbryonic stem cellsInduced pluripotent stem cellsTranscription Factors
*: DDK-tag is the same as FLAG tag. Flag® is a registered trademark of Sigma-Aldrich
DNA-binding activity of GATA4 was measured in OriGene over-expression lysate LY419558 and a control lysate. Three microliters of each lysate was tested with a transcription factor binding assay utilizing GATA4-specific DNA sequences. The high level of activity observed in the over-expression lysate compared to the control lysate demonstrates that the expressed GATA4 is biologically active in the lysate.