mono- and polyubiquitin antibody | FK2
Lanes A and D: K48-linked chains. Lanes B and E: K29-linked chains. Lanes C and F: K63-linked chains.
Mouse anti mono- and polyubiquitin antibody, clone FK2 recognizes both mono- and polyubiquitin chains. The antibody does not recognize 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; Mocciaro and Rape 2012). Lysine-6, Lysine-11, Lysine-27, Lysine-29 and Lysine-33 chains have also been reported (Komander 2009; Ye and Rape 2011).
When comparing staining of mouse anti-mono and polyubiquitin antibody (clone FK2) against staining with mouse anti polyubiquitin antibody (clone FK1) one can determine if a protein target is mono- or polyubiquitinated.
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; Ye and Rape 2011).
When comparing staining of mouse anti-mono and polyubiquitin antibody (clone FK2) against staining with mouse anti polyubiquitin antibody (clone FK1) one can determine if a protein target is mono- or polyubiquitinated.
Product Details
- 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
- Poly-ubiquitinylated lysozyme
- 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. - Guarantee
- 12 months from date of despatch
More Information
- Regulatory
- For research purposes only
Applications of mono- and polyubiquitin antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | |||
| Immunocytochemistry | |||
| Immunoprecipitation | |||
| Western Blotting | 1/100 | 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 mono- and poly-ubiquitin recognizes mono- and poly-ubiquitinylated protein conjugates but does not recognize 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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Secondary Antibodies Available
Product Specific References
Source Reference
-
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.
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