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.
Store freeze-dried powder at 2-8°C. When ready to use, rehydrate with indicated volume of d. water and centrifuge if not clear. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid. Prepare working dilution fresh each day. For extended storage after rehydration, add an equal volume of glycerol (ACS grade or better) for a final concentration of 50%, and store at -20°C as a liquid. Note: after the addition of glycerol, the concentration of protein and buffer salts is one-half of the original. Alternatively, aliquot and freeze the product at -70°C or below in the absence of glycerol. Avoid repeated freezing and thawing.
Expiration date: one year from date of rehydration. However, the expiration date may be extended if the product is stored according to the recommendation and the test results are acceptable for its 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® 647-conjugated antibodies absorb light maximally around 651 nm and fluoresce maximally around 667 nm. They are brighter than Cy5 and DyLight 650 in aqueous mounting media. Alexa Fluor® 647- and APC-conjugated secondary antibodies are the best choice for flow cytometry when secondary antibodies fluorescing at these wavelengths are desired. Alexa Fluor® 647 conjugates are the best choice of far red-emitting dyes for multiple-labeling detection with a confocal microscope.
A significant advantage of using Alexa Fluor® 647 over lower wavelength-emitting dyes is the low autofluorescence of biological specimens in this region of the spectrum. However, because of its peak emission at 667 nm, Alexa Fluor® 647 cannot be seen well by eye, and it cannot be excited optimally with a mercury lamp. Therefore, Alexa Fluor® 647 is not recommended for use with conventional epifluorescent microscopes. It is most commonly visualized with a confocal microscope equipped with an appropriate laser for excitation and a far-red detector. Alexa Fluor® 647 conjugates are less expensive alternatives to allophycocyanin conjugates for flow cytometry.
This product is for IN VITRO RESEARCH USE ONLY. It is not a medical device and it is not intended for diagnostic or therapeutic purposes.
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.