Anti-Ubiquitin (P4D1)

Anti-Ub Mouse Monoclonal Antibody

Additional Information

Product Name: Anti-Ubiquitin (P4D1)
Also Known As: Anti-Ub
Catalog No.: Y3012
Size: 200 µg (2 X 100 µg)
Molecular Weight: ~55/25 kDa
Species: Recognizes ubiquitin in a wide range of species
Source: Immunogen: Full length bovine ubiquitin
Stock: 50 mM PIPES, Ph7.0, 1% sucrose, 0.5% dextran, 0.02% azide and 50% glycerol.
Concentration: 1 mg/ml
Quality Assurance: Guaranteed for detecting 5 ng endogenous ubiquitin or polyubiquitin chains in whole cell or tissue lysates at an antibody dilution of 1:1,000.
Storage: Store the antibody at -20°C. Aliquot is not necessary.
PDF Data Sheet: PDF Datasheet, MSDS
NCBI RefSeq: N/A
Image(s): (Click image to enlarge)


Figure 1.Western blot applications
Anti-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 cells
HeLa 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.

Shipping Method: Wet ice shipping
References: N/A

Details

Specificity
Recognizes mono and polyubiquitinated proteins, free polyubiquitin chains and ubiquitin in a wide range of species. This antibody recognizes different types of polyubiqutin chains.

Isotype
Mouse IgG1

Clone
P4D1

Purification
Protein G affinity chromatography

Applications
WB and IF

Working Conditions
WB (with ECL): 1:1000 ~ 1:3000 dilution
IF: 1:500
IP: not recommended
For best results of WB and IF, and other assays (e.g.: Dot and ELISA), please determine optimal working dilution by titration test

Images
(Click image to enlarge)


Figure 1.Western blot applications
Anti-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 cells
HeLa 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.