Animal Blood B Booster Kit

B Cell Stimulation: Animal Blood B Booster Kit [DDXK-AnBBB] - Microscopic observation of canine lymphocyte activation after the first stimulation.

B Cell Stimulation: Animal Blood B Booster Kit [DDXK-AnBBB] - Microscopic observation of canine lymphocyte activation after 2nd stimulation.

• Reactivity: Mu, Rt, Ca, Eq, Rb, Sh
• Applications: Func

Animal Blood B Booster Kit Summary

• Description: This kit is an extension of the Human Blood B Booster Kit (DDXK-HuBBB) to animal species. Used in combination with the murine myeloma SP2/0 cell line, the Animal Blood B Booster kit (DDXK-AnBBB) is intended for immortalization of B lymphocytes isolated from a wide range of animal species (validated in horse, rabbit, sheep, rat, dog).
• Kit Type: Kit

Applications/Dilutions

• Dilutions:
  • B Cell Stimulation
• Application Notes: This kit is an extension of the Human Blood B Booster Kit (DDXK-HuBBB) to animal species. Used in combination with the murine myeloma SP2/0 cell line, the Animal Blood B Booster kit (DDXK-AnBBB) is intended for immortalization of B lymphocytes isolated from a wide range of animal species (validated in horse, rabbit, sheep, rat, dog).

Packaging, Storage & Formulations

• Storage: Storage of components varies. See protocol for specific instructions.

Notes

This product is manufactured by Eurobio Scientific (formerly Dendritics) and distributed by Novus Biologicals.

Alternate Names for Animal Blood B Booster Kit

• Animal B cell immortalization Animal Blood B Booster
• Animal B cell immortalization
• Animal Blood B Booster kit
• Blood B Booster Kit
• Blood B Booster

Background

Dendritics routinely uses the hybridoma technology described by Kohler and Milstein, where the freshly isolated splenocytes of immunized mice, are fused with the Ig non secreting murine myeloma SP2/0 cell line to immortalize Ig secreting B lymphocytes. The resulting repertoire is generally limited either by the low immunogenicity of the antigen, or by the natural immunity of the mouse. Because the Blood B Booster system (DDXK-HuBBB) was demonstrated as being powerful for human B cells, we attempted to improve the efficiency of the hybridoma technology by introducing an activation step before the fusion in an attempt to give access to a broader repertoire. Briefly, after the immunization program, freshly isolated splenocytes were separately stimulated according to two stimulation systems based on B cell activation through CD40 triggering. We have assumed that the inter-species homology of the CD40 antigen will allow non human B cell activation by a monoclonal antibody raised against human CD40. After the demonstration that the introduction of the stimulation step before fusion significantly increased the clonal diversity of the immortalized murine B cell populations, this process was successfully extended to other animal species starting from peripheral blood samples (Razanajaona-Doll D et al, AAI 98th annual meeting, Immunology 2011TM, San Francisco, CA, USA). The kit AnBBB(R) was packaged so as to make it very easy to handle for the scientists ; the kit AnBBB(R) contains all the reagents needed to obtain around 3.103 independent B cell clones from 6.107 splenocytes or PBMC using six 96 wells culture plates for the stimulations and ten 96 wells plates for the fusion step. The resulting cells are available for a wide range of studies including genomic, transcriptomic and proteomic fields.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.