Cat. No.
MABL-681
Application
crystallization, SPR, ELISA, FC
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
AL-57
From
Recombinant Antibody
Specificity
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.
Alternative Names
LFA-1A; integrin LFA-1; Integrin alpha-L; activated LFA-1 clone 57; CD11 antigen-like family member A; Leukocyte adhesion glycoprotein LFA-1 alpha chain ;Leukocyte function- associated molecule 1 alpha chain
UniProt
P20701
Immunogen
The original antibody was selected from a human Fab-displaying library against the HA open conformation of the LFA-1 I domain.
Application Notes
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).
Antibody First Published
Huang et al. Identification and characterization of a human monoclonal antagonistic antibody AL-57 that preferentially binds the high-affinity form of lymphocyte function- associated antigen- J Leukoc Biol. 2006 Oct;80(4):905-14. doi: 10.1189/jlb.1105649. Epub 2006 Aug 3. PMID:16888085
Note on publication
The original paper describes the generation and characterization of the antibody.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
