Anti β Galactosidase  

Western blotting using Rockland's Fluorescein conjugated anti-b-Galactosidase antibody 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 (p/n MB-070). The membrane was probed with fluorescein conjugated anti-b-Galactosidase (p/n 200-4236) 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 VersaDocR Imaging System developed by BIO-RAD. Other detection systems will yield similar results.

Western blotting using Rockland's anti-b-Galactosidase antibody. 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 4° C with Blocking Buffer for Fluorescent Western Blotting (p/n MB-070). The membrane was probed with anti-b-Galactosidase diluted to 1:10,000. Reaction occurred overnight at 4°C. Dylight649？ conjugated Gt-a-anti-Rabbit IgG (p/n 611-143-120) 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 VersaDocR Imaging System developed by BIO-RAD. Other detection systems will yield similar results.

Western blot using Rockland's anti-b-Galactosidase antibody 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 4° C followed by washes and reaction with a 1:10,000 dilution of IRDyeR 800 conjugated Gt-a-Rabbit IgG (H&L) MX10 (611-132-122) 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). IRDyeR 800 fluorescence image was captured using the OdysseyR Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.