Today's Research.
Tomorrow's Discovery.
Tomorrow's Discovery.
| Datasheet | ||
HeLa whole cell extract lysate
HeLa whole cell extract lysate
| Catalog Number: NB800-PC1 |
Price:
$345.00
|
This ready-to-use whole cell lysate is derived from a cell line or tissue 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.
Research Areas:
Positive Controls and Cell Lysates
Uses:
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 95 degrees 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 0.01-0.03 ml depending on the size format of your gel.
Other applications have not been tested.
Dilutions:
Suggested working dilutions *
Western Blot
* Investigator should determine optimal working dilutions.
Positive Controls:
HeLa is only applicable as a positive control for specific antibodies.
Packaging:
0.8 ml (2.0mg) of HeLa whole cell extract. Concentration determined by Lowry assay.
Concentration:
2.5 mg/ml
Buffer:
1X Laemmeli Sample Buffer
Preservative:
None
Storage:
Store at -80C. Avoid freeze-thaw cycles.
Limitations:
This product is for research use only and is not approved for use in humans or in clinical diagnosis.
Notes:
The cells were grown in DMEM supplemented with 10% FBS. The lysate was prepared by first washing the cells in PBS supplemented with EDTA and a cocktail of protease inhibitors. Washed cells are 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, 0.25% sodium deoxycholic acid to lyse the cells. Protein integrity is ensured using a cocktail of protease inhibitors with broad specificity for the inhibition of aspartic, cysteine, and serine proteases as well as aminopeptidases (0.5 mM AEBSF HCl, 0.4 mM Aprotinin, 25 uM Bestatin, 7.5 uM E-64, 10 uM Leupeptin Hemisulfate and 5 uM Pepstatin A). Cell debris was removed by membrane filtration.
Lane 1: Coommassie stain of 0.02 ml HeLa extract (NB 800-PC1), separated in a 4-20% gradient gel under reducing conditions. Lane 2: Molecular weight standard.
|