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Product name
Cell Fractionation Kit - HT
See all Cell Fractionation products (2) ...
Tests
1 x 96 well plate
Assay type
Quantitative
Assay time
1h 00m
Cross reactivity
Reacts with
Human, a wide range of other species
Product overview
ab109718 (MS862) 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, and thus fractionates adherent cells into cytosol-containing, mitochondria-containing and nuclei-containing fractions. These fractions are referred throughout the protocol as cytosolic, mitochondrial and nuclear fractions. The kit prepares sufficient sample material for subsequent Western blot analyses, dot blot analyses or for analyses by microplate ELISA assays.
ab109718 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, ab109719 (MS861) Cell Fractionation Kit - Standard. This version is designed for fractionating cells grown in batches and is ideal for experiments requiring larger sample amounts.
Notes
Preparation of cytosolic, mitochondrial and nuclear fractions in microplate format.
Storage: 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.
Storage: 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.
Tested applications
Functional Studiesmore details
Properties
Storage instructions
Please see Notes section
| Components | 1 x 96 tests |
|---|---|
| 5X SDS Sample Buffer | 1 x 1.5ml |
| 96-well collection plate with seal film | 3 units |
| Buffer A | 1 x 34ml |
| Detergent HT I | 1 x 7.5ml |
| Detergent HT II | 1 x 35ml |
| Detergent HT III | 1 x 700µl |
Research areas
Kits/ Lysates/ Other >> Kits >> Molecular Biology Kits >> Cell Fractionation Kits
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ELISA - Cell Fractionation Kit - HT (ab109718)
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Alternative names
- MS862
Applications
Show applications key
Our Abpromise guarantee covers the use of ab109718 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Functional Studies
FuncSFuncS
Cell Fractionation Kit - HT images:
ELISA - Cell Fractionation Kit - HT (ab109718)
Qantitative 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 µM Staurosporine (A and B) or with 0.0 and 1.0 µM Staurosporine (C, D, E) were prepared as described in the protocol. Fractions, each derived from one well of a 96-well plate, were analyzed by 'ab110172/ MSA41 Cytochrome c Protein Quantity Microplate Assay Kit (MitoSciences) (A, C and D). Parallel analyses of fractions prepared independently and thus representing inter-assay variation of the ab109718 Cell Fractionation Kit HT are shown in C and D. Western blot analyses of cytochrome c using MSA12 ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail (MitoSciences), 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).
Protocols
References for Cell Fractionation Kit - HT (ab109718)
This product has been referenced in:
- Poster presented at the combined Keystone Symposia Cell Death Pathways and Mitochondrial Dynamics and Physiology, March 22-27, 2009. <a href="http://www.mitosciences.com/PDF/keystone_poster.pdf">Download</a> : (0).Read more (PubMed: ) »
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"