IgG, Library Primer Set, Mouse, BioGenomics(TM)

Many monoclonal antibodies of mouse origin are valuable diagnostic agents. Their production by classical hybridoma techniques is frequently limited by the instability of cell lines, low antibody yields and the limitations of immunizing mice with toxic antigens. A promising alternative to the hybridoma technology is the production of recombinant antibodies. Pioneering work of the last decade showed that it is possible to amplify rearranged immunoglobulin genes from B-lymphocytes, to insert them into different vectors, and to express them in bacteria, yeast, insect, mammalian or plant cells. Moreover, the randomized combination of cloned heavy and light chain immunoglobulin gene fragments allowed the construction of mouse antibody libraries. These libraries enable the isolation of specific antibodies against particular antigens by phage display techniques. One prerequisite for generating highly diversified mouse antibody libraries, however, is the development of PCR primers capable of amplifying all rearranged immunoglobulin genes. In immunoglobulin repertoire library cloning, the homology between a particular primer sequence and its target template, as well as the diversity of a primer pool are the two most important parameters which determine the cloning efficiency and the size of a resulting repertoire library. This screening strategy allows the amplification of rearranged mouse immunoglobulin genes of individual B cell clones as well as of larger B cell populations for the construction of mouse scFv-antibody libraries.\n\nKit Components:\nPrimer Set 1 for PCR Amplification \nI1904-10A1: Heavy Chain Variable Primer (Set 1) 11x20ul \nI1904-10A2: Heavy Chain Constant Primer (Set 1) 1x220ul\nI1904-10A3: Light Chain Variable (kappa) Primer (Set 1) 10x20ul\nI1904-10A4: Light Chain Constant (kappa) Primer (Set 1) 1x220ul\nI1904-10A5: Light Chain Variable (lambda) Primer (Set 1) 1x20ul\nI1904-10A6: Light Chain Constant (lambda) Primer (Set 1) 1x20ul\nPrimer Set 2 for Cloning \nI1904-10B7: Heavy Chain Variable Primer (Set 2)11x20ul \nI1904-10B8: Heavy Chain Constant Primer (Set 2) 1x220ul\nI1904-10B9: Light Chain Variable (kappa) Primer (Set 2) 10x20ul\nI1904-10B10: Light Chain Constant (kappa) Primer (Set 2) 1x20ul\nI1904-10B11: Light Chain Constant (lambda) Primer (Set 2) 1x220ul\nI1904-10B12: Light Chain Constant (lambda) Primer (Set 2) 1x20ul\n\nPCR amplification (Set 1) and cloning (Set 2) of mouse IgG heavy and light chain variable domain coding regions.\n\nApplications:\nGeneration of large repertoires of rearranged immunoglobulin variable domain coding regions for the construction of mouse IgG scFv-antibody libraries. Amplification of immunoglobulin variable gene fragments from single B cell clones.\n\nRecommended Dilution:\nWorking dilution 10pmol/ul (Ready-to-use for common PCR).\nDilution buffer H2O.\nCategory PCR oligonucleotide primer.\n\nStorage and Stability:\nStore at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for 6 months.

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SPECIFICATIONS

Catalog Number

I1904-10A

Size

2x25primers

Form

Supplied as a liquid in 10mM Tris-HCl pH 8, 1mM sodium EDTA.

Purity

Purified by HPLC; high purity, salt-free.

References

1. Zhou, H., et al., Nucleic Acids Research 22: 888-889 (1994). 2. Breitling, F., et al., Gene 104: 147-153 (1991). 3. Du?bel, S., et al., Gene 128: 97-101 (1993).

Additional Info

Determination Mouse IgG Library Primer Set 1 and 2.