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:- 2-5 µg / mlHisto-/Cyto-Chemistry:-2-5 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.
ConjugateCoumarin AMCA
Amax: 350Emax: 450nmAminomethylcoumarin 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.
Images & References
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.
