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F(ab')2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region. F(ab')2 fragments have two antigen-binding Fab portions linked together by disulfide bonds and therefore they are divalent. The average molecular weight is about 110 kDa. They are used for specific applications, such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G.
Extended Storage after Rehydration: Aliquot and freeze at -70°C or below. Avoid repeated freezing and thawing. Alternatively, add an equal volume of glycerol (ACS grade or better) for a final concentration of 50%, and store at -20°C as a liquid.
Expiration date: one year from date of rehydration. The expiration date may be extended if test results are acceptable for the intended use.
1:100 - 1:800 for most applications
Dilution factors are presented in the form of a range because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The actual dilution used must be determined empirically.
Alexa Fluor® 488-conjugated antibodies absorb light maximally at 493 nm and fluoresce with a peak around 519 nm. In aqueous mounting media they are brighter than FITC, Cy2, and DyLight 488. Alexa Fluor® 488 conjugates are recommended for maximum sensitivity for all immunofluorescence procedures requiring a green-fluorescing dye, except for protocols that include mounting in plastic mounting media.
Primary mixed glial culture derived from P0-P3 mouse pups, stained for the astrocytic intermediate filament GFAP (Sigma G3893) and the Gap junction protein Connexin 43 (Santa Cruz sc-9059). Staining was visualized using Jackson ImmunoResearch secondary antibodies donkey-anti-mouse Alexa488 (715-546-150) and donkey-anti-rabbit Cy3 (711-165-152).
Carlyn Martina-Mamber, Elly M. Hol - Netherlands Institute for Neuroscience
This product is provided under an agreement between Life Technologies Corporation and Jackson ImmunoResearch Laboratories, Inc, and the manufacture, use, sale or import of this product is sold pursuant to a license from Life Technologies Corporation for use of its fluorescent dye technology. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell or otherwise transfer this product or its components to any third party, or for any other commercial purposes. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 or firstname.lastname@example.org.