Storage and Rehydration: Store freeze-dried solid at 2-8°C. Rehydrate with the indicated volume of dH2O (see product specification sheet) and centrifuge if not clear. Prepare working dilution on day of use. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid.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.
Buffer: 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6 Stabilizer:15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free) Preservative:0.05% Sodium Azide Suggested Working Concentration or Dilution Range:Western Blot:- 0.5-2 µg / mlHisto-/Cyto-Chemistry:- 0.5-2 µg / mlDilution 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.
ConjugateCyanine Cy™5
Amax: 650Emax: 670nmCy5 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.
Images & References
Above
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.
Below
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.
