Polyubiquitin (K63-LINKAGE Specific) antibody | HWA4C4
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Mouse anti polyubiquitin (K63-linkage specific) antibody clone HWA4C4 recognizes Lysine-63 linked polyubiquitin chains.
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; 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 2009).
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; 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 2009).
Product Details
- Target Species
- Broad
- Species Cross-Reactivity
-
Target Species Cross Reactivity Broad - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by ion exchange chromatography from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- Synthetic peptide containing the ubiquitin K63-isopeptide motif
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Shelf Life
- 12 months from date of despatch
More Information
- Regulatory
- For research purposes only
Applications of Polyubiquitin (K63-LINKAGE Specific) antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| Immunocytochemistry | 1/100 | ||
| Immunohistology - Frozen | 1/100 | ||
| Western Blotting 1 | 1/500 | 1/1000 |
- 1 Specificity for K63 polyubiquitin is temperature sensitive and excessive heat should be avoided in all applications
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 polyubiquitin K63 linkage specific antibody recognizes K63-linked polyubiquitin in a wide range of species but does not recognize any other isopeptide-linked polyubiquitinylated proteins 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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Secondary Antibodies Available
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| TidyBlot™ Western Blot Detection Reagent:HRP | STAR209P | 0.5 ml | WB* |
Product Specific References
References for Polyubiquitin (K63-LINKAGE Specific) antibody
-
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.
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