Cat. No.
MABL-2910
Application
crystallization, IP, NTRL, FC, IF
Isotype
Engineer antibody
Species Reactivity
Reovirus strain T1L
Clone No.
5C6
From
Recombinant Antibody
Specificity
The structure of 5C6 against Sigma1 was solved crystallographically to 3.7 angstrom resolution (Dietrich et al, 2017). The 5C6 Fab regions each contact the upper part of the Sigma1 'head', which forms a beta-spiral repeat structure. The VH CDRs contact the antigen more extensively than the VL CDRs, resulting in a discontinuous binding interface. Reoviruses bind to cell-surface glycans as coreceptors, and in particular T1L reovirus is known to bind the glycan GM2 to facilitate infectivity (Reiss et al, 2012). The 5C6 binding site on Sigma1 partially overlaps with its GM2 binding site, blocking viral infectivity by sterically inhibiting receptor binding. 5C6 bound Sigma1 with an affinity of 2 nM, shown by SPR. Haemagglutination assays of 5C6 showed activity against reovirus strain T1L, but not strain T3D (Stencel-Baerenwald et al, 2015).
Alternative Names
Outer capsid protein sigma1; sigma1 protein; Reovirus strain T1L sigma 1; strain T1L S1; T1L S1; type 1 haemagglutinin; type 1 HA; T1L HA
UniProt
P04506
Immunogen
Murine antibodies were produced by immunising CAF1 mice with reovirus strain T3D, with subsequent immunisation boosts using UV-inactivated strain T1L reovirus. Immunised mice produced antibodies recognising both T3D and T1L. Splenocytes were obtained from immunised mice and fused with the P3Ag8.653 myeloma cell line to generate hybridomas.
Application Notes
5C6 was shown recently to block reoviral infection of L929 mouse fibroblasts in a dose- dependent manner, using immunofluorescence staining (Dietrich et al, 2017). Fibroblasts were incubated with virus-antibody mixtures, and immunofluorescence staining of the cells enabled detection of infectivity. 5C6 is highly specific for the T1L reoviral strain, as opposed to the T3D strain which was also tested. 5C6 was also used to immunoprecipitate reoviral particles.
Antibody First Published
Virgin et al. Monoclonal antibodies to reovirus reveal structure/function relationships between capsid proteins and genetics of susceptibility to antibody action. J Virol. 1991 Dec;65(12):6772-81. PMID:1719233
Note on publication
Describes the generation of a panel of anti-reovirus (strain T3D and T1L) mAbs,including clone 5C6, using mouse immunisation and hybridoma generation. mAb specificities were probed using ELISAs and Western blotting.
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.
