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:- 2-5 µg / ml
Histo-/Cyto-Chemistry:- 2-5 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.
Aminomethylcoumarin Acetate (AMCA) conjugates absorb light maximally around 350 nm and fluoresce maximally around 450 nm. For fluorescence microscopy, AMCA can be excited with a mercury lamp and observed using a UV filter set. Since blue fluorescence is not well detected by the human eye, AMCA-conjugated secondary antibodies should be used only with the most abundant antigens in multiple-labeling experiments. Ways of improving the visibility of AMCA include dark adapting the eyes, using fluorite instead of glass objectives, avoiding mounting media that absorb UV light (such as plastic-based media), and capturing photographic images with blue-sensitive film or CCD cameras. AMCA fades rapidly in conventional epifluorescence and confocal microscopy, and therefore it should be used with mounting media containing an anti-fading agent such as n-propyl gallate.
Connexin expression in astrocytes of middle-aged rat retina
Tissue specimen of 9 month old (middle-aged) Wistar rat retina illustrates connexin expression in astrocytes in the mid-peripheral retina. Six-marker immunohistochemistry was executed on retinal wholemount preparations. Connexin 26 (blue, Alexa532-conjugated anti-goat IgG), Connexin 30 (red, Cy™3 AffiniPure Goat Anti-Rabbit IgG (H+L) - Jackson ImmunoResearch 111-165-003), Connexin 43 (green, Alexa 488-conjugated mouse IgG1), Connexin 45 (pale brown, Alexa 594-conjugated anti-sheep IgG), GFAP stained for astrocytes (pale purple, Cy™5 AffiniPure Donkey Anti-Chicken IgY (IgG) (H+L) - Jackson ImmunoResearch 703-175-155), GS Isolectin B4 stained for blood vessels (turquoise, AMCA-conjugated streptavidin - Jackson ImmunoResearch 016-150-084).
Mansour H, McColm JR, Cole L, Weible M II, Korlimbinis A, et al. (2013) Connexin 30 Expression and Frequency of Connexin Heterogeneity in Astrocyte Gap Junction Plaques Increase with Age in the Rat Retina. PLoS ONE 8(3): e57038. doi:10.1371/journal.pone.0057038
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.