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Figure 1. Quantitative ELISA analysis of cytochrome c release from the mitochondria into the cytosol in HeLa cells induced to undergo apoptosis by Staurosporine treatment.

Cytosolic (C), mitochondrial (M) and nuclear (N) fractions of HeLa cells treated for 4 hrs with 0.00, 0.02, 0.06, 0.18, 0.54, 1.62, 4.86 and 14.58 ÁM Staurosporine (A and B) or with 0.0 and 1.0 ÁM Staurosporine (C, D, E) were prepared using Cell Fractionation Kit HT. Fractions, each derived from one well of a 96-well plate, were analyzed by Cytochrome c Protein Quantity Microplate Assay Kit (MSA41) (A, C and D). Parallel analyses of fractions prepared independently and thus representing inter-assay variation of the Cell Fractionation Kit HT are shown in C and D. Western blot analysis of cytochrome c using ApoTrackÖ Cytochrome c WB Antibody Cocktail (MSA12) is shown for comparison (B and E). Data represent mean +/- standard error of the mean, n=4 (A and C), n=3 (D), n=2 (E), n=1 (B).

Cell Fractionation Kit - HT

Catalog No. MS862

$195.00 - 1 Kit

Product Description

Cell Fractionation Kit HT provides a method and reagents for a simple and rapid preparation of cytosolic, mitochondrial and nuclear fractions from adherent cells. The kit is especially suitable for, but not limited to, high-throughput fractionation of adherent cells in a 96-well plate format. The entire fractionation of 96 samples into the cytosolic, mitochondrial and nuclear fraction is achieved in one hour. The kit is based on sequential detergent extraction of cytosolic, mitochondrial and nuclear proteins without the need for mechanical disruption of cells. The kit prepares sufficient sample material for subsequent Western blot analyses, dot blot analyses or for analyses by microplate ELISA assays.

MS862 is designed to allow the measurement of any proteins which are differentially represented in the cytosol, mitochondria and nuclei, and is particularly applicable to studies of proteins that translocate between these three cellular compartments.

A second version of the kit is also available, MS861 Cell Fractionation Kit - Standard. This version is designed for fractionating cells grown in batches and is ideal for experiments requiring larger sample amounts.

Product Specifications
Kit Components: 34 mL of Buffer A
7.5 ÁL of Detergent HT I
35 ÁL of Detergent HT II
700 ÁL of Detergent HT III
1.5 mL of 5X SDS Sample Buffer.
3 EA of 96-well collection plate and seal film
Applications: Preparation of cytosolic, mitochondrial and nuclear fractions in microplate format.
Species Reactivity: human
Storage Conditions: Store Buffer A, Detergent HT II, Detergent HT III and 5X SDS Sample Buffer at -20░C. Store Detergent HT I at -80░C. Store the collection plates and seal films at room temperature.
Country of Origin: USA

Poster presented at the combined Keystone Symposia Cell Death Pathways and Mitochondrial Dynamics and Physiology, March 22-27, 2009. Download

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