Anti-Human TL1A/TNFSF15 & TNF-alpha Reference Antibody (AMG 966, RUO)

Catalog No.	HF526066
Brand	abinScience
Tested applications	WB, FCM
Species reactivity	Human
Form	Liquid
Storage buffer	0.01M PBS pH 7.4
Purity	>95% purity as determined by SDS-PAGE.
Isotype	IgG1-kappa
Applications	ELISA, Bioactivity: FACS, Functional assay, Research in vivo
Target	TNFSF15, Vascular endothelial cell growth inhibitor, VEGI, TL1, TNF ligand-related molecule 1, Tumor necrosis factor ligand superfamily member 15, TNF, Tumor necrosis factor ligand superfamily member 2, N-terminal fragment, ICD2, NTF, TNF-a, TNF-alpha, Tumor necrosis factor, TNFSF2, TNFA, Cachectin, ICD1
References	1. Kroenke MA, et al. Immune Complex Formation Is Associated With Loss of Tolerance and an Antibody Response to Both Drug and Target. Front Immunol. 2021;12:782788. Published 2021 Dec 14. [HF526066]
Purification	Protein A/G purified from cell culture supernatant.
Biological activity	AMG-966 (300 nM; 6 days) elicits minimal CD4+ T cell proliferation in a human DC:T cell assay from 30 global HLA-representative donors, with no impact from its engineered charge pair mutations [1]. AMG-966 (1:1, 1:1:1, 9:1:1, 200:1:1, 600:1:1 molar ratios; approximately 16 hours) forms intermediate-sized immune complexes with recombinant human TNF at a 1:1 ratio and larger complexes when combined with both TNF and TL1A at lower drug-to-target ratios, as measured by SEC-MALS after 16-hour incubation at 4°C [1]. AMG-966 (1:1:1, 4:1:1, 9:1:1, 100:1:1 molar ratios; overnight) forms immune complexes with recombinant human TNF and TL1A that exhibit enhanced binding to FcγRIa and FcγRIIa at low 1:1:1 drug-to-target ratios (after overnight incubation at 4°C), with binding decreasing as the drug-to-target ratio increases, while FcγRIIIA binding is minimally affected [1].
Endotoxin level	< 10 EU/mg
Expression system	Mammalian cells
Accession	O95150 & P01375
Stability and Storage	Use a manual defrost freezer and avoid repeated freeze thaw cycles. Store at 2 to 8°C for frequent use. Store at -20 to -80°C for twelve months from the date of receipt.
Alternative name	Bispecific, AMG966, AMG-966
Clone ID	AMG 966
Note	For research use only. Not suitable for clinical or therapeutic use.
Isotype Control	Human IgG1 Recombinant Isotype Control Antibody (13R4) (HV080507)

Western blot	Western blot analysis was performed using anti-TNFSF15/TL1A monoclonal antibody at 1ug/mL on various samples. Lane 1: recombinant human TNFSF15/TL1A (Catalog No: HF526011) Lane 2: negative control
SDS-PAGE	SDS-PAGE for Anti-Human TNFSF15/TL1A & TNFa/TNF-alpha Reference Antibody (AMG 966, RUO)
Flow CytoMetry	Flow-cytometry APC using anti-human TNFa antibody. Untransfected cells (blue Histogram) and Transfected cells (Yellow Histogram) were stained with an APC anti-human TNFa monoclonal antibody (Catalog: HF526066) at a concentration of 5 µg/ml for 30 mins at RT. After washing, cells analysed on a NovoCyte Flow Cytometer.
Flow CytoMetry	Flow-cytometry FITC using anti-human TNFa antibody. Untransfected cells (blue Histogram) and Transfected cells (Yellow Histogram) were stained with an FITC anti-human TNFa monoclonal antibody (Catalog: HF526066) at a concentration of 5 µg/ml for 30 mins at RT. After washing, cells analysed on a NovoCyte Flow Cytometer.
Flow CytoMetry	Flow-cytometry using anti-human TNFa antibody. 293T Transtected cells were stained with an irrelevant antibody (Blue Histogram) or an anti-human TNFa monoclonal antibody (Catalog: HF526066, Yellow Histogram) at a concentration of 5 µg/ml for 30 mins at RT. After washing, bound antibody was detected using a a Goat Anti-Human IgG H&L Polyclonal Antibody, FITC (abinScience: HF690414) and cells analysed on a NovoCyte Flow Cytometer.
Bioactivity	Detects recombinant human TNFa (Cat No.: HF879011) in indirect ELISA.
Bioactivity	Detects recombinant human TNFa (Cat No.: HF526011) in indirect ELISA.
Bioactivity	Detects Recombinant Human TNFSF15/TL1A Protein, C-His (Cat No.: HF526011) in indirect ELISA.