Microsome Isolation Kit (BN00514)

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SKU:
BN00514
  • Microsome Isolation Kit (BN00514)
  • Microsome Isolation Kit (BN00514)
€607

Description

ELISA Kit Technical ManualMSDS

Microsome Isolation Kit

Microsomes are spherical vesicle-like structures formed from membrane fragments following homogenization and fractionation of eukaryotic cells. The microsomal subcellular fraction is prepared by differential centrifugation and consists primarily of membranes derived from the endoplasmic reticulum (ER) and Golgi apparatus. Microsomes isolated from liver tissue are used extensively in pharmaceutical development, toxicology and environmental science to study the metabolism of drugs, organic pollutants and other xenobiotic compounds by the cytochrome P450 monooxidase (CYP) enzyme superfamily. Microsomal preparations are an affordable and convenient in vitro system for assessing Phase I biotransformation reactions, as they contain all of the xenobiotic-metabolizing CYP isozymes and the membrane-bound flavoenzymes (such as NADPH P450-Reductase and cytochrome b5) required for function of the multicomponent P450 enzyme system. Assay Genie's Microsome Isolation Kit enables preparation of active microsomes in about one hour, without the need for ultracentrifugation or sucrose gradient fractionation. The kit contains sufficient reagents for 50 isolation procedures, yielding microsomes from roughly 25 grams of tissue or cultured cells.

Figure: (a) Western blot analysis of microsomal and S9 fractions isolated from rat liver. Microsomes and S9 fraction were isolated according to the kit protocol described above. A total of 30 µg of protein in SDS-PAGE buffer was loaded in each lane and run on a 4- 20% gradient gel. The blots were probed for Cytochrome P450 (CYP2E1) (Cat. no. # 3084R), mitochondrial marker VDAC1 (Cat. no. #3594) and ER-specific protein marker calnexin (Cat. no. # 3811). Blots show enrichment of CYP and calnexin and depletion of mitochondrial membrane proteins in microsomal fraction. (b) Relative densitometry data demonstrate the enrichment of Cytochrome P450 and reduction of mitochondrial protein marker in microsomal fraction as compared to whole rat liver homogenate (each column shows mean density ± SEM relative to whole liver lysate for at least 2 repeats).

Key Information Description

Product SKU

BN00514

Size

50 assays

Application

  • Convenient and fast isolation of microsomal fraction from animal tissues
  • Assessment of CYP-mediated drug metabolism and xenobiotic biotransformation
  • Protein profiling of microsomal membrane proteins by SDS-PAGE and Western blot

Features and Benefits

  • Simple & convenient protocol
  • The kit provides unique formulations of ready-to-use buffers and reagents to isolate microsomes without the need for ultracentrifugation or sucrose gradient fractionation

Kit Components

  • Homogenization Buffer
  • Storage Buffer
  • Protease Inhibitor Cocktail

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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