Catalogue

Background
Total cell Iysates are useful as both positive and negative controls in immunoblotting. A431, a human epidermoid carcinoma cell line, and the EGF-stimulated A431 Iysates are used as negative and positive controls, respectively, when studying the phosphorylation cascade initiated by ligand binding to receptor tyrosine kinases. Both A431 and normal human fibroblasts are valuable positive controls for antibodies to proteins expressed in these cells. The mouse fibroblast cell line 3T3 and its RSV-transformed counterpart serve as negative and positive controls, respectively, for antibodies to proteins that are phosphorylated as a result of transformation by RSV (Rous sarcoma virus). Madin-Darby bovine kidney (MDBK) cell Iysate displays proteins expressed in bovine kidney and PC12 (rat phaeochromocytoma) cell Iysate can be expected to mirror the protein expression of cells and tissues of neuroectodermal origin. Likewise, Jurkat cells express the proteins unique to human T-cells. Rat brain is a widely used positive control for those proteins that are exclusively expressed in brain. At right is an example of the use of positive and negative control Iysates in western blotting. The blot of 3T3 cell Iysate (lane 1) and RSV-transformed 3T3 (lane 2) was probed with anti-phosphotyrosine peroxidase conjugate to reveal those proteins that are tyrosine-phosphorylated in response to transformation by

Synonyms: 3T3 Cell Lysate

Product

Product Form: Unconjugated

Formulation: Provided as solution in Laemmli electrophoresis sample buffer. Samples need to be boiled for 3-5 minutes, before loading the gel.

Concentration: See vial for concentration

Applications
Suggested use: A volume of 10-25ul is suggested for immunobloting.

Functional Analysis: Western Blotting

Storage
Product should be stored at -20°C. Aliquot to avoid freeze/thaw cycles

Product Stability: See expiration date on vial

Shipping Conditions: Room Temperature

Caution
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but our company accepts no liability for any inaccuracies or omissions in this information.