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U-2OS Whole Cell Lysate

CAT: 0247-OCRA00099-500UGSize: 500 µgDry Ice: NoHazardous: No
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CAT#:0247-OCRA00099-500UGSize:500 µg
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24/48H Stock Items & 2 to 6 Weeks non Stock Items.
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Description
Ready-to-use whole cell lysates are derived from cell lines or tissues using highly refined extraction protocols to ensure exceptionally high quality, protein integrity and lot-to-lot reproducibility. All extracts are tested by SDS-PAGE using 4-20% gradient gels and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.
Reactivity
Human
Conjugation
Unconjugated
Type
Cell Lysate
Applications
Sodium sodecyl sulfate - polyacrylamide gel electrophoresis, Western blot
Assay Protocol
Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol
Concentration
1 mg/ml
Format
Liquid// 1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% glycerol, 0.005% bromophenol blue pH 8.8) with (v/v) 10% glycerol
Reconstitution
Store vial at -70° C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles. Expiration date is three (3) months from date of opening if stored at -20° C or (1) year from date of opening if stored at -70° C.
Specificity
The 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 uM Bestatin, 1.5 µM E-64, 2 uM Leupeptin Hemisulfate, 1 uM 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.

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