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C2C12 Whole Cell Lysate

C2C12 Whole Cell Lysate

Catalog Number: orb348725

DispatchUsually dispatched within 5-10 working days
$ 289.00
Catalog Numberorb348725
CategoryTools
DescriptionC2C12 Whole Cell Lysate
Tested applicationsChIP, IP, SDS-PAGE, WB
Concentration4.0 mg/ml
Dilution rangeChIP: User Optimized, IP: User Optimized, WB: User Optimized
Form/AppearanceLiquid (sterile filtered)
PurityC2C12 cells were grown in Dulbecco's medium supplemented with 10% fetal bovine serum. Cells were washed with PBS and then incubated on ice in modified RIPA buffer, containing 150 mM sodium chloride, 50 mM Tris HCl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid , 0.1% SDS and 0.01% (w/v) sodium azide to lyse the cells. Protein integrity was ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.1 mM AEBSF HCl, 0.08 µM Aprotinin, 5 µM Bestatin, 1.5 µM E-64, 2 µM Leupeptin Hemisulfate, 1 µM Pepstatin A). Phosphatase inhibitors 1 mM NaF and 1 mM Na3VO4 were also added. Cell debris was removed by centrifugation. Protein concentration was determined by a modified Lowry assay using a commercially available kit. Protein concentration was adjusted to 4 mg/ml with 1X RIPA buffer.
ConjugationUnconjugated
Hazard InformationNon-Toxic
SourceMouse
StorageStore vial at -70° C or COLDER. For extended storage, aliquot lysate to minimize freeze/thaw cycles.
Buffer/PreservativesPreservative: None. Stabilizer: None. 1X RIPA Buffer with HALT Protease and Phosphatase Inhibitors
Alternative namesC2C12 Lysate, Cell Lysate
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NoteFor research use only
Application notesorb348725 has been tested by SDS-PAGE. Multi-purpose C2C12 Whole Cell Lysate are especially prepared as positive control for multiple assays including western blot, immunoprecipitation (IP), capture ELISA or other assays requiring native protein sample. For separation by SDS-PAGE and subsequent western blot analysis, lysates should be diluted by user to desired concentration in SDS-PAGE buffer with 2-mercaptoethanol or dithiothreitol as the reducing agent and heated to 95° C for 5 minutes. Sample is ready for use in immunoprecipitation and ELISA experiments, conditions should be optimized by the user. Rockland recommends its TrueBlot IP reagents for immunoprecipitation experiments.
Expiration Date12 months from date of receipt.
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