U-138 MG Whole Cell Lysate

Images

 
SDS-Page: U-138 MG Whole Cell Lysate [NBP3-06999] - SDS-PAGE Results of U-138 MG Whole Cell Lysate. Lane 1: U-138 MG Whole Cell lysate(10ug) Reduced. Lane 2: Opal Prestained Molecular Weight Marker. Lane 3: U-138 MG ...read more

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB, PAGE

Order Details

U-138 MG Whole Cell Lysate Summary

Preparation
Method
The U-138 MG 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 uM Aprotinin, 5 uM Bestatin, 1.5 uM 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 SDS 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.
Purity
Multi-step

Applications/Dilutions

Dilutions
  • SDS-Page
  • Western Blot
Application Notes
This product has been tested by SDS-PAGE. Ready-to-use lysates 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 95C 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 uL depending on the size format of your gel.

Packaging, Storage & Formulations

Storage
Store at -70C. Avoid freeze-thaw cycles.
Buffer
1X SDS-PAGE Sample Buffer (62.5 mM Tris HCl, 2% SDS, 10% Glycerol and 0.005% bromophenol blue, pH 6.8), 10% (v/v) Glycerol
Preservative
No Preservative
Purity
Multi-step

Lysate Details for Array

Type
Cell
Subcellular Fraction
Whole

Notes

Store vial at -70C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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Video Protocols

WB Video Protocol

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Array NBP3-06999

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