Cat. No.
MABL-1162
Application
FACS, SPR, ELISA, FC
Isotype
Engineer antibody
Species Reactivity
Human, Rhesus Monkey
Clone No.
BL006-19-29G5-29K1
From
Recombinant Antibody
Specificity
This antibody is specific for the D2 domain of the shorter form of human CD96 (isoform v2). It was found to also bind the long for of human CD96, albeit with lower affinity, and to cross-react with Rhesus CD96.
Alternative Names
Tactile; T cell activation, increased late expression; Cluster of Differentiation 96
UniProt
P40200
Immunogen
The original antibody was generated by immunizing wild-type mice with human CD96 DNA, followed by a boost with CHO cells transfected with the human CD96 expression vector.
Application Notes
The original antibody (Mouse IgG2a) was selected using ELISA and flow cytometry to confirm binding to CD96. FACS testing showed that this antibody exhibits strong binding to the human CD96 short isoform (CD96v2) expressed on CHO cells (EC50 = 2.98 nM) and strong but weaker binding to the human CD96 long isoform (CD96v1). This antibody showed partial inhibition of the CD96/CD155 interaction, although it did not achieve 50% inhibition at the maximum tested concentration (30 μg/mL, 200 nM), with a relative IC50 calculated at 5.24 nM. FC analysis revealed strong binding to T, NK, and NKT cells from human peripheral blood mononuclear cells (PBMCs). SPR measurement revealed a KD of 0.182 nM towards an in- house produced hCD96-hFc protein. Similar to BL006-19-14, this antibody co-stimulated T cell activation and proliferation in the presence of low concentrations of anti-CD3, as evidenced by increased CD25 expression and CFSE dilution. Its co-stimulatory effect was dependent on Fc receptor engagement. In tumor-infiltrating lymphocytes (TILs) from head and neck cancer patients, the antibody promoted TIL proliferation, particularly when baseline proliferation was low. The antibody exhibited optimal co-stimulation when added simultaneously with suboptimal CD3 activation (US20230192845A1).
Antibody First Published
Note on publication
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.
