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Catalog Number | orb348689 |
---|---|
Category | Antibodies |
Description | HT-29 Lystate |
Clone Number | HT-29 |
Tested applications | IP, SDS-PAGE, WB |
Concentration | 4.0 mg/ml |
Dilution range | IP: User Optimized, WB: User Optimized |
Form/Appearance | Liquid |
Purity | HT-29 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 2 mg/ml and then an equal volume of 2X SDS-PAGE sample buffer was added. |
Conjugation | Unconjugated |
Hazard Information | Non-Toxic |
Source | Human |
Storage | Store HT-29 Whole Cell Lysate at -70° C or COLDER. For extended storage, whole cell lysate to minimize freeze/thaw cycles. |
Buffer/Preservatives | 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) |
Alternative names | HT29 Lysate, Cell Lysate, HT-29 lysate Read more... |
Note | For research use only |
Application notes | ready-to-use HT-29 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. |
Expiration Date | 12 months from date of receipt. |
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