| Description | |
|---|---|
| Overview | The ProteoExtract® Tissue Dissociation Buffer Kit is a tool for isolation of viable cells from fresh tissue. The isolated cells can then be used for total, subcellular, membrane, or other form of proteome extractions. This kit serves as an accessory product for the following kits: ProteoExtract® Subcellular Proteome Extraction Kit (Cat. No.: 539790), ProteoExtract® Subcellular Proteome Extraction Kit, Mini (Cat. No.: 539791), ProteoExtract® Native Membrane Protein Extraction Kit (Cat. No.: 444810). |
| Catalogue Number | 539720 |
| Brand Family | Calbiochem® |
| Application Data |   Tissue dissociation of rat pancreatic tissue and human pancreatic tissue was carried out according to the protocol outlined above. 1.3 x 107 dissociated cells were extracted using the ProteoExtract® Subcellular Proteome Extraction Kit protocol (Cat. No. 539790), incorporating the protocol changes as outlined above. Proteins from the cytosolic fraction, the membrane fraction, the nuclear fraction, and the cytoskeletal fraction were separated by SDS-PAGE, transferred to PVDF membrane, and probed with antibodies to the indicated markers. |
| Materials Required but Not Delivered | • Collagenase: the collagenase should efficiently and specifically dissociate the tissue type of choice. Based on internal testing, a minimum of 10,000 units (~8-10 mg) collagenase is recommended per gram of tissue. The exact amount will depend on the specific enzyme and type of tissue. The list of enzymes below represents possible options used successfully with this kit. Please always refer to the manufacturer's recommendations and specifications for use. Table 1: Collagenases Used With Various Tissues • Kit for downstream processing (optional; e.g. ProteoExtract® Subcellular Proteome Extraction Kit, Cat. No. 539790, ProteoExtract® Subcellular Proteome Extraction Kit Mini, Cat. No. 539791, or ProteoExtract® Native Membrane Protein Extraction Kit, Cat. No. 444810) • Hemocytometer • Phosphate Buffered Saline (PBS) • Pasteur or micropipet • Microscope • Trypan blue (e.g. 0.18% in PBS) • 15 ml conical centrifuge tubes • Thermo mixer • Sieve • Scalpel or scissors • Refrigerated centrifuge (minimum rcf = 350 g) • Petri dish • 37°C incubator with gentle agitation |
| Product Information | |
|---|---|
| Form | 10 Dissociation reactions, 1 g each |
| Kit contains | 10X Tissue Dissociation Buffer, Protease Inhibitor Cocktail, Tissue Dissociation Buffer Additive and a user protocol. |
| Quality Level | MQ400 |
| Biological Information | |
|---|---|
| Sample Type | Fresh tissue samples from eukaryotic organisms |
| Product Usage Statements | |
|---|---|
| Intended use | The Calbiochem® ProteoExtract® Tissue Dissociation Buffer Kit is a tool for isolation of viable cells from fresh tissue for the purpose of protein extraction. It is an accessory product for the following kits: • ProteoExtract® Subcellular Proteome Extraction Kit (S-PEK, Cat. No. 539790) • ProteoExtract® Subcellular Proteome Extraction Kit, Mini (S-PEK Mini, Cat. No. 539791) • ProteoExtract® Native Membrane Protein Extraction Kit (M-PEK, Cat. No. 444810). With the exception of the collagenase enzyme the set contains all reagents required for efficient dissociation of tissue samples. The goal of the cell isolation procedure is to maximize the yield of viable dissociated cells, so the end-user must select the collagenase appropriate for each tissue with respect to the type of tissue, the species of origin, and the age of the animal. A list of enzymes is provided below in Materials Required but Not Provided. |
| Storage and Shipping Information | |
|---|---|
| Ship Code | Ambient Temperature 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 | +2°C to +8°C |
| Storage Conditions | Upon arrival store the entire contents of the kit at 4°C. Following reconstitution, all components are stable for a minimum of 2 days at 4°C or for up to 2 months at -20°C. Avoid freeze/thaw cycles of reconstituted solutions. |
| Do not freeze | Ok to freeze |
| Supplemental Information | |
|---|---|
| Kit contains | 10X Tissue Dissociation Buffer, Protease Inhibitor Cocktail, Tissue Dissociation Buffer Additive and a user protocol. |
