beta Galactosidase antibody (F

Western blot using GeneTex anti-beta Galactosidase antibody (GTX26641) shows detection of a band at ~117 kDa (lane 1) corresponding to b-Gal present in a partially purified preparation (arrowhead). Approximately 1μg of protein was resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking, the membrane was probed with the primary antibody diluted to 1:1,000. Reaction occurred overnight at 4º C followed by washes and reaction with a 1:10,000 dilution of IRDyeR 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers in lane M (700 nm channel, red). IRDye 800 fluorescence image was captured using the Odyssey Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.

Western blotting using GeneTex Fluorescein conjugated anti-b-Galactosidase antibody (GTX26641) shows a band at ~117 kDa (lanes 1 - 3) corresponding to 60 ng, 30 ng and 15 ng, respectively of b-Gal present in partially purified preparations (arrowhead). Lane 4 shows no cross reactivity with proteins present in a non-specific control E.coli lysate. Proteins were resolved on a 4-20% Tris-Glycine gel by SDS-PAGE and transferred to nitrocellulose and blocking using Blocking Buffer for Fluorescent Western Blotting. The membrane was probed with fluorescein conjugated anti-b-Galactosidase (GTX26641) diluted to 1:10,000. Reaction occurred for 2 hours at room temperature. Molecular weight estimation was made by comparison to a prestained MW marker in lane M.Fluorescence image was captured using the VersaDoc Imaging System developed by BIO-RAD. Other detection systems will yield similar results.

Western blotting using GeneTex anti-beta-Galactosidase antibody (GTX26641). Lane 1 shows 80 ng and lane 2 shows 20 ng loaded onto gel. Results for non-reducing conditions of SDS-PAGE prior to transfer to nitrocellulose are shown on the left side of the figure; results obtainined under reducing conditions are shown on the right. Blots were blocked overnight at 4º C with Blocking Buffer for Fluorescent Western Blotting. The membrane was probed with anti-b-Galactosidase diluted to 1:10,000. Reaction occurred overnight at 4ºC. Dylight649™ conjugated Gt-a-anti-Rabbit IgG was used for detection. Molecular weight estimation was made by comparison to a prestained MW marker (center).in lane M. Fluorescence image was captured using the VersaDoc Imaging System developed by BIO-RAD. Other detection systems will yield similar results.