Mouse Lung Whole Tissue Lysate (Adult Whole Normal)

SDS-Page: Mouse Lung Whole Tissue Lysate (Adult Whole Normal) [NBP3-05286] - Lane 1: Mouse Lung, Adult, Whole Cell Lysate (10ug) Reduced. Lane 2: Opal Prestained Molecular Weight Markers Lane 3: Mouse Lung, Adult, Whole Cell Lysate (10ug) Reduced. Results show wide range of molecular weight bands.

• Reactivity: Mu
• Applications: WB, PAGE

Mouse Lung Whole Tissue Lysate (Adult Whole Normal) Summary

• Description: Store vial at -70C or COLDER. For extended storage, aliquot contents to minimize freeze/thaw cycles.
• Preparation Method: Tissues were washed exhaustively with PBS to remove blood and other debris. A lysate was prepared by homogenizing the tissue and washing the cells in cold PBS. Washed cells were incubated at 4C in modified RIPA buffer containing 150 mM sodium chloride, 50 mM Tris Cl, pH 7.4, 1 mM EDTA, 1.0% NP-40, 0.5% sodium deoxycholic acid and 0.1% SDS 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 (0.1 mM AEBSF HCl, 0.08 uM Aprotinin, 5 uM Bestatin, 1.5 uM E-64, 2 uM Leupeptin Hemisulfate and 1 uM Pepstatin A). The following phosphatase inhibitors were also added: 1 mM NaF and 1 mM Na3VO4. Cell debris was removed by centrifugation and membrane filtration. Protein concentration was determined by BCA assay using a commercially available kit. The 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: Tissue
• Life Stage: Adult
• Tissue Condition: Normal
• Subcellular Fraction: Whole

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.