CD104 (Integrin beta4)

Integrin beta 4 (also known as GP150 and CD104) is a 150-200kD member of the Integrin beta family. It forms noncovalent heterodimers with Integrin alpha 6 and participates in the formation of epithelial hemidesmosomes. Human Integrin beta 4 is a type I transmembrane glycoprotein that is 1795 amino acids (aa) in length. It contains a 683 aa extracellular domain (ECD) (aa # 28-710) and a large 1089 aa cytoplasmic region. Two ECD alternate splice forms may exist. One shows an 84 aa substitution for the 84 aa between aa # 621-704. A second shows a 144 aa substitution for the first 704 amino acids. Human Integrin beta4 ECD shares 88% sequence identity with mouse Integrin beta 4 ECD.\n\nApplications: \nSuitable for use in ELISA, Western Blot, Immunocytochemistry and Flow Cytometry. Other applications have not been tested. \n\nRecommended Dilutions: \nWestern Blot: 0.1-0.2ug/ml. The detection limit is ~1ng/lane under non-reducing and 10ng/lane under reducing conditions.\nImmunocytochemistry: 10ug/ml detects Integrin B4 in human A431 cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 min. at RT and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 min. at RT. After blocking, cells were incubated with diluted C2446-01F overnight at 4 degrees C followed by Rhodamine RedTM-coupled anti-sheep IgG at RT in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. \nDirect ELISA: 0.5-1.0ug/ml. The detection limit is ~0.3ng/well. \nFlow cytometry: 50ug/ml. Add 10ul to 1-2.5x10e5 cells in a total reaction volume not exceeding 200ul. \nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at -20 degrees C. Stable for 12 months at -20 degrees C. Reconstitute by adding sterile 40-50% glycerol, PBS. Aliquot and store at -20 degrees C. Reconstituted product is stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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