Cat. No.
MABL-3120
Application
functional assays, in vivo, SPR, ELISA
Isotype
Engineer antibody
Species Reactivity
Human
Clone No.
Nb70
From
Recombinant Antibody
Specificity
This antibody is specific for human TNFR1.
Alternative Names
TNFRI; Tumor necrosis factor RI; Tumor necrosis factor R1; Tumor necrosis factor receptor type 1-associated death domain protein; TNFR1-associated death domain protein; TNFRSF1A-associated via death domain; Nb 14; Nanobody 14
UniProt
Q15628
Immunogen
The original antibody was generated by immunizing an alpaca (Vicugna pacos) with soluble human TNFR1 (hTNFR1). A VHH library was constructed using total RNA from peripheral blood lymphocytes, and panning on hTNFR1 was performed to isolate antigen-specific nanobodies.
Application Notes
The original format of this antibody (VHH) was used in ELISAs to determine its binding affinity to soluble human TNFR1 (hTNFR1) and other proteins such as albumin, human TNFR2 (hTNFR2), and mouse TNFR1 (mTNFR1); a KD of 7.709 nM was measured for hTNFR1. This antibody was tested in a HEK-2 Blue assay to evaluate its inhibitory effect on the signaling of human TNF (hTNF) through TNFR1; it was found to effectively inhibit hTNFR1 by competing with TNF for binding with an IC50 value of 4.144 μM and with lymphotoxin alpha (LTa) with an IC50 of 220 nM. This antibody's affinity for hTNFR1 was evaluated by SPR analysis; a KD of 38.71 nM was measured. This antibody was used to create a trivalent nanobody by linking it to clones Nb 96 and Nb Albumin using flexible [G4-S]3 linkers, and the resulting nanobody (Nb Alb- 70-96) — TNF Receptor-One Silencer (TROS) — was tested for various in vitro, in vivo, and ex vivo (Steeland et al., 2015; PMID: 25538244).
Antibody First Published
Steeland et al. Generation and characterization of small single domain antibodies inhibiting human tumor necrosis factor receptor 1 J Biol Chem. 2015 Feb 13;290(7):4022-37. doi:10.1074/jbc.M114.617787 PMID:25538244
Note on publication
The original publication describes the generation and characterization of a selective human TNFR1 inhibitor, named TROS (TNF Receptor-One Silencer), based on Nanobody (Nb) technology.
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.
