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.
Western Blot:- 0.5-2 µg / ml
Histo-/Cyto-Chemistry:- 0.5-2 µg / ml
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.
Cy5 conjugates are excited maximally at 650 nm and fluoresce maximally at 670 nm. They can be excited to about 98% of maximum with a krypton/argon laser (647 nm line) or to about 63% of maximum with a helium/neon laser (633 nm line). Cy5 can be used with a variety of other fluorophores for multiple labeling due to a wide separation of its emission from that of shorter wavelength-emitting fluorophores. However, we recommend Alexa Fluor® 647 as the preferred far-red-fluorescing dye because it is brighter than Cy5 in aqueous mounting media. The reverse is true when mounting in non-polar, plastic mounting media, i.e. Cy5 is brighter than other far-red-fluorescing dyes, including Alexa Fluor® 647 and DyLight 650.
Jejunum villi superficial epithelium labeled with Ulex-biotin
Streptavidin-Cy5 (016-170-084 blue), gastric K cells labeled with goat anti-GIP + donkey anti-goat-Cy2 (705-225-147 green) and nerve fibers labeled with mouse anti-tubulin + donkey anti-mouse-Cy3 (715-165-151 red).
Brian McAdams and William Kennedy, University of Minnesota.
Vascular-associated astrocytes in the young adult rat retina
Tissue specimen of 3 month old (young adult) Wistar rat retina illustrates vascular-associated astrocytes in the mid-peripheral retina. Triple label immunohistochemistry was executed on retinal wholemount preparations. GFAP stained for astrocytes (red, Cy3-conjugated mouse monoclonal anti-GFAP), GS Isolectin B4 stained for blood vessels (blue, Cy5-conjugated streptavidin - Jackson ImmunoResearch 016-170-084), and S-100 stained for cell soma region of astrocytes (green, Alexa 488-conjugated anti-rabbit IgG).
Mansour, H., Chamberlain, C. G., Weible II, M. W., Hughes, S., Chu, Y. and Chan-Ling, T. (2008), Aging-related changes in astrocytes in the rat retina: imbalance between cell proliferation and cell death reduces astrocyte availability. Aging Cell, 7: 526–540. doi: 10.1111/j.1474-9726.2008.00402.x
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.
Cy is a trademark of GE Healthcare Bio-Sciences Ltd. Jackson ImmunoResearch is licensed by GE Healthcare Bio-Sciences Ltd. to manufacture and sell conjugates of Cy2, Cy3, and Cy5 reactive dyes under US Patent Number 5,268,486, and other patents pending.