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Fab Preparation

  • General scheme for Fab preparation
  • Example 1: Size exclusion HPLC and MALDI-TOF MS analysis of Fab produced from mouse monoclonal antibody (mAb) and rabbit polyclonal antibody (pAb) at CellMosaic
Price:
Please contact us for a quote.
SKU:
BM0006


Product Description

In the preparation of antibody-enzyme conjugates there are a few advantages of using antibody fragments rather than intact antibodies. For example, antibody fragments lacking an Fc region have less interference with various Fc binding proteins, can easy penetrate cell membranes due to their lower molecule weight, and experience lower nonspecific binding to cell surfaces. Fab fragments are useful in immuno-histochemical studies.

  • Chemistry:  Cleavage of IgG molecule in the hinge region above the interchain disulfides using papain to create two identical Fab portions and one intact Fc fragment.
  • Specification: Pure Fab.
  • Procedure: After a buffer exchange (if necessary) digestion of IgG, in-process gel electrophoresis analysis, purification, and concentration determination.
  • Price: Contact us for a quote.
  • Starting materials: Please check biopolymer for ordering information.  
  • Note: Due to the complexity of the digestion, differences of IgG subclasses and species of origin, extra charges may be added for some antibodies if additional procedures or test runs are deemed necessary.

Examples (see images):

  1. Fabs produced from mouse monoclonal antibody (mAb) and Rabbit polyclonal antibody (pAb) at CellMosaic (shown below): high quality Fab based on size exclusion HPLC, gel electrophoresis (not shown here), and MALDI-TOF MS analysis.bm0006-fab-copy.jpg

References:

  1. Rousseaux, J.; Biserte, G.; Bazin, H. The differential enzyme sensitivity of rat immunoglobulin G subclasses to papain and pepsin. Molecular Immunology 1980, 17, 469-482.
  2. Coulter, A.; Harris, R. Simplified preparation of rabbit Fab fragments. J. Immun. Methods 1983, 59, 199-203.
  3. Rousseaux, R.; Rousseaux-Prevost, R.; Bazin, H. Optimal conditions for the preparation of Fab and F(ab')2 fragments from monoclonal IgG of different rat IgG subclasses. J. Immun. Methods 1983, 64, 141-146.

 

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