Pyruvate Kinase (PKLR) (NM_000298) Human Over-expression Lysate

Specifications

Product Data
Description	Transient overexpression lysate of pyruvate kinase, liver and RBC (PKLR), nuclear gene encoding mitochondrial protein, transcript variant 1
Species	Human
Expression Host	HEK293T
Expression cDNA Clone or AA Sequence	TrueORF Clone RC206455
Tag	C-Myc/DDK
Detection Antibodies	Clone OTI4C5, Anti-DDK (FLAG) monoclonal antibody (TA50011-100)
Predicted MW	61.8 kDa
Components	1 vial of 100 µg gene specific transient over-expression cell lysate in RIPA buffer 1 vial of 100 µg whole HEK293T cell lysate in RIPA buffer 1 vial of 250ul 2xSDS Sample Buffer (4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT)
Storage	The lysate is shipped with dry ice. Upon receiving, store the sample at -80°C. Also after dilution, the protein sample should be aliquoted and stored at -80°C for long term storage. Avoid repeated freeze-thaw cycles. Lysate samples can be diluted with 2xSDS Sample Buffer provided. Lysate samples are stable for 12 months from the date of receipt when stored at -80°C.
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.).
Reference Data
RefSeq	NP_000289
Locus ID	5313
Cytogenetics	1q22
Protein Families	WB
Protein Pathways	Glycolysis / Gluconeogenesis, Insulin signaling pathway, Maturity onset diabetes of the young, Metabolic pathways, Purine metabolism, Pyruvate metabolism, Type II diabetes mellitus

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