Anti-Ub Mouse Monoclonal Antibody

Additional Information

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Anti-Ubiquitin (P4D1)

Anti-Ub

Y3011

100 µg

~55/25 kDa

Recognizes ubiquitin in a wide range of species

Immunogen: Full length bovine ubiquitin

lyophilized powder

lyophilized powder

Guaranteed for detecting 5 ng endogenous ubiquitin or polyubiquitin chains in whole cell or tissue lysates at an antibody dilution of 1:1,000.

1) Store the lyophilized antibody desccated at -20°C. 2) After reconstitution, store the antibody at -20°C. Aliquot is not necessary. The quality of the antibody is guaranteed for one year after puchasing if stored correctly.

PDF Datasheet

N/A

(Click image to enlarge)Figure 1.Western blot applicationsAnti-Ubiquitin (P4D1) was used at a 1:500 dilution. Bovine thymus ubiquitin was run as follows; Lane 1-50 ng, Lane 2-25 ng. Lane 3-12.5 ng, Lane 4-6.25 ng, Lane 5-3.12 ng. Lanes 6 & 7 represent 20 μg of Swiss 3T3 cell lysate from cells treated for 5h with 10 µM MG-132 (Lane 6) or untreated cells (Lane 7). Arrow indicates free ubiquitin band (8 kD), higher molecular weight bands are ubiquitinated proteins.Figure 2. IF of HeLa cellsHeLa cells were stained and visualized by fluorescence microscopy. Ubiquitin targeted cytoplasmic and nuclear proteins and free ubiquitin were stained using Anti-Ubiquitin (P4D1) at 1:500 dilution.

Room temperature shipping

N/A

Details

SpecificityRecognizes mono and polyubiquitinated proteins, free polyubiquitin chains and ubiquitin in a wide range of species. This antibody recognizes different types of polyubiqutin chains. IsotypeMouse IgG1 CloneP4D1 PurificationProtein G affinity chromatographyApplicationsWB and IFWorking ConditionsWB (with ECL): 1:1000 ~ 1:3000 dilutionIF: 1:500IP: not recommendedFor best results of WB and IF, and other assays (e.g.: Dot and ELISA), please determine optimal working dilution by titration testReconstitutionCentrifuge briefly to collect the powder to the bottom of the tube prior to reconstitution. Reconstitute each 100 μg antibody into 100 μL 50% glycerol in miliQ water. Final buffer composition is 50 mM PIPES, Ph7.0, 1% sucrose, 0.5% dextran and 50% glycerol. Add 0.02% azide to prevent bacterial cotamination. Images(Click image to enlarge)Figure 1.Western blot applicationsAnti-Ubiquitin (P4D1) was used at a 1:500 dilution. Bovine thymus ubiquitin was run as follows; Lane 1-50 ng, Lane 2-25 ng. Lane 3-12.5 ng, Lane 4-6.25 ng, Lane 5-3.12 ng. Lanes 6 & 7 represent 20 μg of Swiss 3T3 cell lysate from cells treated for 5h with 10 µM MG-132 (Lane 6) or untreated cells (Lane 7). Arrow indicates free ubiquitin band (8 kD), higher molecular weight bands are ubiquitinated proteins.Figure 2. IF of HeLa cellsHeLa cells were stained and visualized by fluorescence microscopy. Ubiquitin targeted cytoplasmic and nuclear proteins and free ubiquitin were stained using Anti-Ubiquitin (P4D1) at 1:500 dilution.