CloneID: AL-57
Heavy Chain modification: His-Tagged
Antigen Long Description: The original antibody was selected from a human Fab-displaying library against the HA open conformation of the LFA-1 I domain.
Buffer Composition: PBS with 0.02% Proclin 300.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: P20701
Specificity Statement: The antibody binds to the alpha L I domain in a HA but not LA conformation.The antibody binding site overlaps the ICAM-1 binding site on the I domain.
Application Notes (Clone): The specificity of the original format of the antibody was confirmed by ELISA analysis. The cell-binding specificity of the original and IgG1 format of the antibody was determined using flow cytometric analysis with HA and LA cells, the antibody bound to HA cells in a dose-dependent manner but did not bind to LA cells. The antibody preferentially binds to the active conformation of LFA-1 and blocks LFA-1-mediated adhesion and lymphocyte proliferation (Huang et al., 2006; PMID: 16888085). The original format of the antibody could discriminate among low-affinity, intermediate-affinity (IA), and HA states of LFA-1. Furthermore, the binding of the antibody depended on the presence of Mg2+. The antibody showed no binding to the WT I domain, intermediate binding to the IA I domain (KD = 4.7 μM), and good binding to the HA I domain by SPR (KD = 0.023 μM). The original format of the antibody showed affinity increases on a subset (≈10%) of lymphocyte cell surface LFA-1 molecules upon stimulation with CXC chemokine ligand 12 (CXCL-12). The IgG1 format of the antibody could bind to HA I domain expressed on the surface of K562 cells presence of Mg2+, as tested by using immunofluorescent flow cytometry (Shimaoka et al., 2006; PMID: 16963559). The crystal structures of the Fab fragment of the antibody alone and in complex with IA were determined (Zhang et al., 2009; PMID: 19805116).
English
Deutsch
