Anti-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 683aa extracellular domain (ECD) (aa28-710) and a large 1089aa cytoplasmic region. Two ECD alternate splice forms may exist. One shows an 84aa substitution for the 84aa between aa621-704. A second shows a 144aa substitution for the first 704aa. Human Integrin beta4 ECD shares 88% sequence identity with mouse Integrin beta 4 ECD. Applications: Suitable for use in Direct ELISA, Western Blot, Immunocytochemistry and Flow Cytometry. Other applications have not been tested. Recommended Dilutions: Western Blot: 0.1-0.2ug/ml. The detection limit is ~1ng/lane under non-reducing and 10ng/lane under reducing conditions., Immunocytochemistry: 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°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. , Direct ELISA: 0.5-1.0ug/ml. The detection limit is ~0.3ng/well. , Flow Cytometry: 50ug/ml. Add 10ul to 1-2.5x10e5 cells in a total reaction volume not exceeding 200ul. , Optimal dilutions should be determined by each laboratory for each application. Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. 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.