Cat. No.
MABL-2290
Application
neutralize, SPR, ELISA, FC
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
3F12.2D3
From
Recombinant Antibody
Specificity
3F12 binds to human LTα3 and LTαβ (with a KD of ~0.3 nM, determined by BIACORE; ~ 37 pM, determined by ELISA) in both its soluble homotrimeric and membrane heterotrimeric forms. LTα belongs to the tumor necrosis factor (TNF) superfamily and is secreted as a homo-trimer (LTα3), or is expressed on the cell surface in complex with LTβ. Lymphotoxin is produced by lymphocytes and in its homotrimeric form binds to TNFRSF1A/TNFR1, TNFRSF1B/TNFBR and TNFRSF14/HVEM. LTα3 signaling induces target cells to upregulate many chemokines and cytokines in an NFkB-dependent manner.
Alternative Names
LTα3 chimeric 3F12;LT;; LTα; Tumor necrosis factor beta; Tumor necrosis factor β; Tumor necrosis factor-beta; tumor necrosis factor-β; TNF beta; TNF-beta; TNF β; TNF-β; Lymphotoxin-alpha; Lymphotoxin alpha
UniProt
P01374
Immunogen
3F12 was prepared by hyperimmunizing BALB/c mice with purified recombinant human LTα expressed in E. coli - the hybridoma clone was selected for its ability to bind LTα3 by ELISA. The CDRs of the antibody were grafted to the framework regions of the mice mAb 2G7 to create the chimeric version of the antibody.
Application Notes
The properties of the chimieric 3F12 is equivalent to that of the original 3F12 mAb. 3F12 is able to bind to Th2 cells and additionally is can functionally block human LTα3 versus an isotype control (trastuzumab) - LTα3-induced ICAM upregulation is blocked on HUVEC (Human Umbilical Endothelial Cells) (IC50 0.04 nM) and LTα3-induced IL-8 is blocked in A549 cells. The antibody has also been used in ELISA studies to detect LT-alpha and to measure binding.
Antibody First Published
Catellana et al. Resurrection of a Clinical Antibody: Template ProteoGenomic de novoProteomic Sequencing and Reverse Engineering of an Anti-Lymphotoxin Alpha Antibody Proteomics. 2011 Feb; 11(3): 395–405. PMID:21268269
Note on publication
Describes the generation of anti-LT antibody and compares binding specificity and affinity of the chimeric antibody against the original antibody by BIACORE and ELISA.
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.
