Components

Mitochondria Extraction Buffer, Cytosol Extraction Buffer, Protease Inhibitor Cocktail, Reducing Reagent, and a user protocol.

General description

This kit provides a unique combination of reagents useful for the isolation of a highly enriched mitochondrial fraction from the cytosol. The enriched fractions can be used to study factors of interest using Western blotting, ELISA, or other assays.

Note: 1 Test = reagents sufficient for processing 5 x 10⁷ cells.

Legal Information

PROTEOEXTRACT is a registered trademark of Merck KGaA, Darmstadt, Germany

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Due to the nature of the Hazardous Materials in this shipment, additional shipping charges may be applied to your order. Certain sizes may be exempt from the additional hazardous materials shipping charges. Please contact your local sales office for more information regarding these charges.

Packaging

1 kit in Fibre case

Preparation Note

Protease Inhibitor Cocktail: Reconstitute the lyophilized Protease Inhibitor Cocktail by adding 250 µl DMSO; mix well before use. Dispense into aliquots and store at -20°C. 1X Cytosol Extraction Buffer Mix: Prepare 1X Cytosol Extraction Buffer by diluting the Cytosol Extraction Buffer 1:4 with ddH₂O. Prepare only the amount needed for the assay (5 x 10⁷ cells require 1 ml 1X Cytosol Extraction Buffer Mix). Just prior to use add 2 µl Protease Inhibitor Cocktail and 1 µl DTT to 1 ml 1X Cytosol Extraction Buffer. Mitochondria Extraction Buffer Mix: Prepare only the amount needed for the assay (5 x 10⁷ pelleted cells require 100 µl Mitochondria Extraction Buffer Mix). Just prior to use, add 2 µl Protease Inhibitor Cocktail and 1 µl DTT to 1 ml of Mitochondria Extraction Buffer.

Principle

The ProteoExtract™ Cytosol/Mitochondria Fractionation Kit is designed for extraction of mammalian cells to yield enriched mitochondrial and cytosolic fractions for use in downstream analyses, such as Western blotting.

Storage and Stability

Upon arrival store the entire contents of the kit at -20°C. Following initial use, store the buffers at 4°C and the DTT and Protease Inhibitor Cocktail at -20°C.

Warning

Toxicity: Multiple Toxicity Values, refer to MSDS (O)