Anti-Lymphotoxin alpha [3F12.2D3]

• Catalog Number: orb613927
• Category: Antibodies
• Description: Mouse monoclonal antibody to Lymphotoxin alpha
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 3F12.2D3
• Tested applications: ELISA, FC
• Reactivity: Human
• Isotype: IgG2b
• 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.
• Concentration: 1 mg/ml
• Purity: Purified
• Conjugation: Unconjugated
• Target: Lymphotoxin alpha
• UniProt ID: P01374
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: LTα3 chimeric 3F12;LT;; LTα; Tumor necrosis factor
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow-cytometry using the anti-Lymphotoxin alpha antibody 3F12.2D3 U937 cells were fixed using 2% PFA, permeabilised using 0.5% Triton and stained with unimmunized rabbit IgG antibody (MOPC-21; isotype control, black line) or the rabbit IgG-chimeric version of 3F12.2D3 (orb613926, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Western Blot using anti-Lymphotoxin alpha antibody 3F12.2D3 Lipopolysaccharide (LPS)-treated human peripheral blood monocyte (PBM) lysate (35 µg protein in RIPA buffer) was resolved on a 15% SDS PAGE gel and blots probed with the rabbit IgG-chimeric version of 3F12.2D3 (orb613926) at 2 µg/ml before detection using an anti-rabbit secondary antibody. Protein was detected by chemiluminescence. The expected unmodified running size for Lymphotoxin alpha is 22.3kDa, though the form shown here is likely glycosylated. orb613926 successfully detected Lymphotoxin alpha in LPS-treated human PBM samples.