| Description | |
|---|---|
| Overview | 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. | |
| Catalogue Number | QIA88 |
| Brand Family | Calbiochem® |
| Application Data |  Mitochondrial and cytosolic fractions were isolated from HEPG2 cells using QIA88 ProteoExtract® Cytosol/ Mitochondria Fractionation Kit. Mitochondrial pellets were resuspended in PBS. The protein concentration of each fraction was determined using the BCA assay. DTT was omitted from the Cytosol Extraction Buffer as it can interfere with concentration determination in the BCA assay. Equivalent amounts (10 µg) of HEPG2 homogenate (total protein), cytosolic, and mitochondrial fractions were analyzed by Western blot for proteins localized to different cellular regions as indicated. Representative results from 2 independent fractionations are shown. |
| Materials Required but Not Delivered | • Phosphate buffered saline (PBS) • Dounce tissue homogenizer • DMSO (for reconstitution of the Protease Inhibitor Cocktail) |
| Product Information | |
|---|---|
| Form | 100 Extractions |
| Format | Cell extraction |
| Kit contains | Mitochondria Extraction Buffer, Cytosol Extraction Buffer, Protease Inhibitor Cocktail, Reducing Reagent, and a user protocol. |
| Biological Information | |
|---|---|
| Sample Type | Cells |
| Safety Information | |
|---|---|
| R Phrase | R: 23/24/25-32-34-36/37/38-41 Toxic by inhalation, in contact with skin and if swallowed. Contact with acids liberates very toxic gas. Causes burns. Irritating to eyes, respiratory system and skin. Risk of serious damage to eyes. |
| S Phrase | S: 23-26-36/37/39-45 Do not breathe fumes. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable protective clothing, gloves and eye/face protection. In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible). |
| Product Usage Statements | |
|---|---|
| Intended use | 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 Shipping Information | |
|---|---|
| Ship Code | Dry Ice Only |
| Toxicity | Multiple Toxicity Values, refer to MSDS |
| Hazardous Materials Attention: | 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. |
| Storage | -20°C |
| Storage Conditions | 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. |
| Avoid freeze/thaw | Avoid freeze/thaw |
| Do not freeze | Ok to freeze |
| Supplemental Information | |
|---|---|
| Kit contains | Mitochondria Extraction Buffer, Cytosol Extraction Buffer, Protease Inhibitor Cocktail, Reducing Reagent, and a user protocol. |
