Products:Cell Biology >> Cell Cycle >> Cell Cycle Inhibitors >> Rb
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ab24 has been referenced in 7 publications.
Publishing research using ab24? Please let us know so that we can cite the reference in this datasheet
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Anti-Rb antibody [Rb1 1F8] - Nuclear Marker (ab24) at 1/1000 dilution + Human Fibroblast cell lysate(nuclear). 16 hour incubation.
Secondary
NIF 825 (provided in ECL kit) - HRP conjugated
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 105 kDa
Observed band size : 100-150 kDa (why is the actual band size different from the predicted?)
Exposure time : 1 minute
This image is courtesy of an Abreview submitted on 5 October 2005.
Predicted band size : 105 kDa
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 for 1 hour. After detection of the protein bands, the blot was stripped and re-probed with mouse monoclonal antibody against β-actin to confirm equal loading. After secondary antibody incubation, the blot was washed and detected by ECL kit in autoradiography.
Image from Lee DY et al, PLoS One 5:e13599 (2010), Fig 3.
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