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A431 Whole Cell Lysate

CAT: 0013-GTR19350904Size: 500 µgDry Ice: NoHazardous: No
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CAT#:0013-GTR19350904Size:500 µg
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24/48H Stock Items & 2 to 6 Weeks non Stock Items.
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Description
A431 Whole Cell Lysate
Product Name Alternative
A431 Lysate, Cell Lysate, A423
Conjugation
Unconjugated
Source
Human
Origin
Human
Applications
WB: User Optimized
Concentration
1.0 mg/ml
Purity
A423 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 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 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added.
Form
Liquid
Storage Conditions
Store A431 Whole Cell Lysate at -70° C or COLDER. For extended storage, whole cell lysate to minimize freeze/thaw cycles.
Notes
For research use only.
Applications Notes
Ready-to-use A431 Whole Cell Lysate are especially prepared as positive controls for separation by SDS-PAGE and subsequent western blot analysis. Lysates are prepared in denaturing buffer WITHOUT dissociating agents (i.e. no 2-mercaptoethanol or dithiothreitol has been added) . Heat lysate to 95°C for 5 minutes and rapidly cool. If dissociating conditions are desired, add reducing agent prior to heating. The recommended loading volume per lane is 10-20 μl depending on the size format of your gel.
Tested Applications
SDS-PAGE, WB
Preservative
Preservative: None. Stabilizer: 10% (v/v) Glycerol. 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8)

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