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Fusion protein. Rb-b-galactosidase fusion protein spanning nucleotides 1126-1973 of human Rb gel purified from bacterial lysates (R. Grand et al. 1989).
Our Abpromise guarantee covers the use of ab24 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 105 kDa.|
|IP||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
Lysates prepared from lung and kidney of VerUTR transgenic and wildtype mice were analyzed by Western Blot probed with ab24.
Increased expression of Rb1 was detected in the organs from the VerUTR transgenic mice. Cells were seeded onto 6-well plates at 2×105 cells per well overnight. They were then transfected with 1 µg of VerUTR or control vector in combination with scrambled RNA or siRNA against VerUTR. Proteins were extracted 48 hours after transfection by lysing in 60 µl of lysis buffer containing protease inhibitors (150 mM NaCl, 25 mM Tris-HCl, pH 8.0, 0.5 mM EDTA, 1% Triton X-100, 8 M Urea, and 1x protease inhibitor cocktail). Tissues were disrupted in appropriate volume of lysis buffer depending on tissue weight. All samples were subjected to SDS-PAGE and then transferred to nitrocellulose membranes followed by incubating with ab24 at 1/500 dilution at 4°C overnight. The secondary antibody used was goat anti-mouse IgG at 1/2000 dilution at room temperature.
Primary antibody was incubated for 16 hours.
Blocked with 5% milk for 1 hour.
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