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™3
Amax: 550Emax: 570nmCy3 is brighter, more photostable, and gives less background than other orange-red fluorescing dye conjugates. Cy3 conjugates can be excited maximally at 550 nm, with peak emission at 570 nm. For fluorescence microscopy, Cy3 can be visualized with traditional tetramethyl rhodamine (TRITC) filter sets, since the excitation and emission spectra are nearly identical to those of TRITC. We recommend Cy3 as a brighter alternative to TRITC. Cy3 can be excited to about 50% of maximum with an argon laser (514 nm or 528 nm lines), or to about 75% of maximum with a helium/neon laser (543 nm line) or mercury lamp (546 nm line). Cy3 has been used with fluorescein for double labeling; however, the use of a narrow band-pass emission filter for fluorescein is recommended to minimize Cy3 fluorescence in the FITC filter set. Cy3 can also be paired with Alexa Fluor® 647 for multiple labeling when using a confocal microscope. However, a better choice for multiple labeling is Rhodamine Red-X because its fluorescence is midway between a green fluorescing dye (like Alexa Fluor® 488) and a far-red-fluorescing dye like Alexa Fluor® 647.
Images & References
Above
Pax2 expression in astrocytes of aged rat retina
Tissue specimen of 19 month old (aged) Wistar rat retina illustrates Pax2 expression in retinal astrocytes in the central retina. Triple label immunohistochemistry was executed on retinal wholemount preparations. Pax2 stained for surrounding nuclei of astrocytes (red, biotinylated anti-rabbit Ig, Cy3-conjugated streptavidin - Jackson ImmunoResearch 016-160-084), DAPI stain for cell nuclei (blue), and GFAP stained for astrocytes (green, Alexa 488-conjugated anti-mouse IgG1).
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
Below
Confocal micrograph of the ventral horn in the D9 spinal cord segment of a red-eared turtle (Trachemys scripta elegans). The composite image of an electrophysiologically measured and biocytin-injected motor neuron (red, Cy3 016-160-084), in a population of motor neurons (green, Choline acetyltransferase, Alexa Fluor® 488 705-545-147) with a nuclear staining (blue, DAPI). Kristian Reveles Jensen, University of Copenhagen.
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.
