Pan Ubiquitin antibody | P4D1

Mouse anti Pan Ubiquitin

Product Type
Monoclonal Antibody
Clone
P4D1
Isotype
IgG1
Specificity
Pan Ubiquitin

Product Code Applications Pack Size List Price Your Price Qty
MCA6040
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SDS Safety Datasheet SDS
IC IP WB 50 µg loader
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loader
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Mouse anti pan ubiquitin antibody, clone P4D1 recognizes both mono- and polyubiquitin chains. The antibody has also been reported to recognize free polyubiquitin chains and free ubiquitin.

Ubiquitin, as the name implies, is a ubiquitously expressed and highly conserved protein of 8.6 kDa. The protein is covalently linked to selected lysine residues in a post-translational modification process known as ubiquitylation or ubiquitination. This chemical reaction is mediated by three different protein families; ubiquitin-activating enzymes (also known as E1s), ubiquitin-conjugating enzymes (also known as E2s) and ubiquitin ligases (also known as E3s) (Hershko and Ciechanover 1998).

The impact of ubiquitination depends on whether a single ubiquitin moiety (monoubiquitination) or an ubiquitin chain (polyubiquitination) has been attached to a protein. Monoubiquitination tends to trigger cellular processes related to endocytosis and membrane trafficking (Haglund et al. 2003) while the impact of polyubiquitination varies depending on how the ubiquitin residues in the chain have been linked. Attachment of Lysine-48 ubiquitin chains results in degradation by the 20S proteasome while addition of Lysine-63 ubiquitin chains mediates DNA damage and NFkappaB signaling (Chen 2005 and Mocciaro and Rape 2012). Lysine-6, Lysine-11, Lysine-27, Lysine-29 and Lysine-33 chains have also been reported (Komander 2009 and Ye and Rape 2011).

When comparing staining of mouse anti pan ubiquitin antibody (clone P4D1) against staining with mouse anti polyubiquitin antibody (clone FK1) one can determine, if a protein target is mono- or polyubiquitinated. In contrast to mouse anti mono- and polyubiquitin antibody (FK2), clone P4D1 also recognizes free ubiquitin.

Target Species
Broad
Product Form
Purified IgG - liquid
Preparation
Purified IgG prepared by ion exchange chromatography from tissue culture supernatant
Buffer Solution
Phosphate buffered saline
Preservative Stabilisers
<0.1% Sodium Azide (NaN3)
Immunogen
Denatured bovine ubiquitin
Approx. Protein Concentrations
IgG concentration 1.0 mg/ml
Regulatory
For research purposes only
Guarantee
12 months from date of despatch

This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.

Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.

This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Application Name Verified Min Dilution Max Dilution
Immunocytochemistry
Immunoprecipitation
Western Blotting 1/1000
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
Mouse anti pan ubiquitin recognizes mono- and poly-ubiquitin protein conjugates, free polyubiquitin chains and free ubiquitin by Western Blot. Use of milk based blocking reagents is not recommended. 1% BSA in PBS or TBS Tween should be used instead.

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References for Pan Ubiquitin antibody

  1. Fujimuro, M. et al (1994) Production and characterization of monoclonal antibodies specific to multi-ubiquitin chains of polyubiquitinated proteins.
    FEBS Lett. 349 (2):173-80.
  2. Wang, H. et al. (2008) Analysis of nondegradative protein ubiquitylation with a monoclonal antibody specific for lysine-63-linked polyubiquitin.
    Proc Natl Acad Sci U S A. 105 (51): 20197-202.

Further Reading

  1. Kahana, A and Gottschling, DE. (1999) DOT4 links silencing and cell growth in Saccharomyces cerevisiae.
    Mol Cell Biol. 19 (10): 6608-20.

Western Blotting

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Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"

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