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Whole IgG antibodies are isolated as intact molecules from antisera by immunoaffinity chromatography. They have an Fc portion and two antigen binding Fab portions joined together by disulfide bonds and therefore they are divalent. The average molecular weight is reported to be about 160 kDa. The whole IgG form of antibodies is suitable for the majority of immunodetection procedures and is the most cost effective.
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.
DyLight 405-conjugated secondary antibodies are excited maximally at about 400 nm and fluoresce with a peak at about 421 nm. They are very bright and photostable, but their optimal use is limited to microscopes or flow cytometers equipped with a 405 nm laser and a 420 nm emission filter. Under these conditions, it is possible to perform effective 4-color imaging with good color separation, good photostability, and high sensitivity. The combination of DyLight 405, Alexa Fluor® 488, Rhodamine Red-X, and Alexa Fluor® 647 provides for maximum color separation. Another 4-color dye combination, which may be equally effective but has slightly less color separation, is DyLight 405, Alexa Fluor® 488, Cy3, and Alexa Fluor® 647. DyLight 405 conjugates are an excellent choice for blue-fluorescing secondary antibodies in multi-color labeling protocols.
DyLight™ is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.