CloneID: M3-8
Antigen Long Description: Callithrix jacchus marmosets were used, and experimental allergic encephalomyelitis was induced by the injection of rat MOG (aa1-125) into the marmosettes. The rMOG was expressed in E. coli and purified to homogeneity. The animals were killed 4-70 days after the onset of symptoms of EAE. Bone marrow and spleen cells were obtained from an immunized C. jacchus, the RNA extracted with Trizol reagent and rtPCR used to generate cloning inserts containing Fab portions of IgGk. Phage display was then used to select for the MOG-reactive Fab fragments using the pCOMB3H phage display vector, and binding confirmed using an ELISA.
Origin Pub PMID: 12060766
Buffer Composition: PBS with 0.02% Proclin 300.
Chimeric Use Statement: This chimeric rat antibody was made using the variable domain sequences of the original Marmoset Fab format, for improved compatibility with existing reagents, assays and techniques.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: Q63348
Specificity Statement: Recognises an epitope which is distinct from M26, M38, M45, M3-24 and M3-31. Is able to displace native anti-MOG Abs from C. jacchus serum. Has been shown to bind in situ by immunofluorescence. The epitope recognised is a structural epitope, shown by the lack of recognition of linear MOG peptides. Has also been shown to compete with anti-MOG Abs from three patients with MS. Does not bind to MOG expressed in CHO cells.
Application Notes (Clone): This antibody is part of a family of antibodies including clones M26, M38, M45, M3-8, M3-24, M3-31. This antibody has been proposed for the diagnosis and prognosis of multiple sclerosis (MS) or experimental allergic encephalomyelitis (EAE) which is a disease model for MS. This is achieved by using competition assays to determine if there are autoantibodies present in an individual which recognise structural epitopes on MOG which have been shown to be associated with the progression of MS. This antibody has been used in ELISAs and competition assays to characterise its epitope (see specificity statement) and determine whether the similar epitopes are recognised in marmosets with EAE and humans with MS. It was originally generated and tested as a Fab (von Budingen et al, 2002). Whilst this has been shown to not bind to hMOG, it has been shown to compete with IgG from MS patients for binding to rMOG (Lalive et al, 2006). Analysis has also been done on the amino acid sequence of this antibody (von Budingen et al, 2006).
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