mono- and polyubiquitin antibody | FK2

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Mouse anti mono- and polyubiquitin

Product Type
Monoclonal Antibody
Clone
FK2
Isotype
IgG1
Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
MCA6035 E IC IP WB 50 µg
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.

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.
Shelf Life
12 months from date of despatch

More Information

Regulatory
For research purposes only

Applications of mono- and polyubiquitin antibody

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
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.

Secondary Antibodies Available

Description Product Code Pack Size Applications List Price Quantity
Human anti Mouse IgG1:HRP HCA036P 0.1 mg E
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed) STAR117A 0.5 mg E WB
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed) STAR117D488GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®549 (Multi Species Adsorbed) STAR117D549GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):DyLight®649 (Multi Species Adsorbed) STAR117D649GA 0.1 mg F IF
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed) STAR117D680GA 0.1 mg F WB
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed) STAR117D800GA 0.1 mg F IF WB
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed) STAR117F 0.5 mg F
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed) STAR117P 0.5 mg E WB
Goat anti Mouse IgG (Fc):FITC STAR120F 1 mg C F
Goat anti Mouse IgG (Fc):HRP STAR120P 1 mg E WB
Rabbit F(ab')2 anti Mouse IgG:RPE STAR12A 1 ml F
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed) STAR13B 1 mg C E P RE WB
Goat anti Mouse IgG:FITC (Rat Adsorbed) STAR70 0.5 mg F
Goat anti Mouse IgG:RPE (Rat Adsorbed) STAR76 1 ml F
Goat anti Mouse IgG:HRP (Rat Adsorbed) STAR77 0.5 mg C E P
Goat anti Mouse IgG/A/M:Alk. Phos. STAR87A 1 mg C E WB
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) STAR87P 1 mg E
Rabbit F(ab')2 anti Mouse IgG:Dylight®800 STAR8D800GA 0.1 mg F IF WB
Rabbit F(ab')2 anti Mouse IgG:FITC STAR9B 1 mg F

Product Specific References

Source Reference

  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.
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