Polyubiquitin (K63-LINKAGE Specific) antibody | HWA4C4

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Mouse anti Polyubiquitin (K63-LINKAGE Specific)

Product Type
Monoclonal Antibody
Clone
HWA4C4
Isotype
IgG2a
Product Code Applications Pack Size List Price Quantity
MCA6032
Datasheet
C IC WB*
MSDS
50 µl loader

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).

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
0.09% 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.
Guarantee
12 months from date of despatch

More Information

Regulatory
For research purposes only

Applications of Polyubiquitin (K63-LINKAGE Specific) 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
Immunocytochemistry 1/100
Immunohistology - Frozen 1/100
Western Blotting 1 1/500 1/1000
  1. 1Specificity 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.

Secondary Antibodies Available

Description Product Code Applications Pack Size List Price Quantity
Human anti Mouse IgG2a:FITC HCA037F F 0.1 mg loader
Human anti Mouse IgG2a:HRP HCA037P E 0.1 mg loader
Goat anti Mouse IgG (H/L):Alk. Phos. (Multi Species Adsorbed) STAR117A E WB 0.5 mg loader
Goat anti Mouse IgG (H/L):DyLight®488 (Multi Species Adsorbed) STAR117D488GA F IF 0.1 mg loader
Goat anti Mouse IgG (H/L):DyLight®680 (Multi Species Adsorbed) STAR117D680GA F WB 0.1 mg loader
Goat anti Mouse IgG (H/L):DyLight®800 (Multi Species Adsorbed) STAR117D800GA F IF WB 0.1 mg loader
Goat anti Mouse IgG (H/L):FITC (Multi Species Adsorbed) STAR117F F 0.5 mg loader
Goat anti Mouse IgG (H/L):HRP (Multi Species Adsorbed) STAR117P E WB 0.5 mg loader
Goat anti Mouse IgG (Fc):FITC STAR120F C F 1 mg loader
Goat anti Mouse IgG (Fc):HRP STAR120P E WB 1 mg loader
Rabbit F(ab')2 anti Mouse IgG:RPE STAR12A F 1 ml loader
Rabbit F(ab')2 anti Mouse IgG:HRP (Human Adsorbed) STAR13B C E P RE WB 1 mg loader
Goat anti Mouse IgG:FITC (Rat Adsorbed) STAR70 F 0.5 mg loader
Goat anti Mouse IgG:RPE (Rat Adsorbed) STAR76 F 1 ml loader
Goat anti Mouse IgG:HRP (Rat Adsorbed) STAR77 C E P 0.5 mg loader
Goat anti Mouse IgG/A/M:Alk. Phos. STAR87A C E WB 1 mg loader
Goat anti Mouse IgG/A/M:HRP (Human Adsorbed) STAR87P E 1 mg loader
Rabbit F(ab')2 anti Mouse IgG:Dylight®800 STAR8D800GA F IF WB 0.1 mg loader
Rabbit F(ab')2 anti Mouse IgG:FITC STAR9B F 1 mg loader

Useful Reagents Available

Description Product Code Applications Pack Size List Price Quantity
TidyBlot Western Blot Detection Reagent:HRP STAR209P WB* 0.5 ml

Application Based External Images

Western Blotting

Product Specific References

References for Polyubiquitin (K63-LINKAGE Specific) antibody

  1. 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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