CD11a (LFA-1, alpha L Integrin)

Integrin subunit aL/CD11a is a 180 kD type I TM glycoprotein that interacts only with integrin b2/CD18 to form LFA-1, a leukocyte adhesion protein which binds endothelial cell ICAM. Human integrin aL (Accession # P20701) contains a 1064 aa extracellular domain (ECD), a 20 aa TM sequence and a 58 aa cytoplasmic domain. The ECD contains seven repeats that form a beta-propeller structure and one inserted vWA domain (I domain) containing a metal ion-dependent adhesion site (MIDAS). Human and mouse integrin aL ECD share 74% aa identity. A second isoform has a 53 aa insert in the ECD. Hybridoma: Produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified CHO-derived, recombinant human Integrin alpha L (rhITGAL; aa 26-1089). Applications: Flow cytometry Western blot Recommended Dilution: Flow cytometry-This antibody has been validated in PBMCs by flow cytometry. Dilute this antibody to 25ug/mL and add 10ul of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding 10 uL of a 25ug/mL stock solution of a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome. Western blot-: 1-2ug/mL with the appropriate secondary reagents to detect human ITGAL. Using a colorimetric detection system, the detection limit for rhITGAL is approximately 20 ng/lane under non-reducing and reducing conditions. Chemiluminescent detection will increase sensitivity by 5 to 50 fold. Optimal dilutions to be determined by the researcher. Storage and Stability: Lyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees 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.