Product Description

The anti-diglycine lysine antibody (anti-K-?-G-G antibody) is produced by immunizing rabbits with conjugates of K-?-GG-KLH. Antibodies are purified by diglycine lysine agarose affinity chromatography. The product specifically recognizes proteins and peptides with Gly-Gly sequences that are present as an adduct on the ?-amine of lysine. This antibody has been well utilized as affinity reagents for proteomic screening of lysine ubiquitinated peptides from trypsin-digested proteins both in eukaryotic and prokaryotic cells.

Source/Purification

The anti-diglycine lysine antibody (anti-K-?-G-G antibody) is produced by immunizing rabbits with conjugates of K-?-GG-KLH. Antibodies are purified by diglycine lysine agarose affinity chromatography.

Specificity

All

Recommended Applications

IAC ?IAC=Immunoaffinity Immunoprecipitation

?

Recommended antibody dilution

/

Important

Only for IP-MS,not for WB or other application.

? ? ?

A. Dot blotting?analysis on indicated amount of K-?-GG-KLH (lane 1), K-?-GG peptide library (lane 2), ?synthetic unmodified human histone H3 peptide STGGKAPR (aa 10-17) (lane 3), ?KLH (lane 4) and control peptide of K-?GG peptide library (lane 5, unmodified lysine peptide ?library) using anti-diglycine lysine rabbit pAb (1:2000).

?

B. Mass spectrometer analysis: Representative annotated MS/MS spectra of an ?ubiquitin remnant-containing peptide.? Crude proteins from ?HeLa?cell?lysate?were digested ?by trypsin.?The ubiquitin remnant-containing peptides were?affinity ?enriched by the?anti-diglycine lysine rabbit pAb?followed by LC-ESI-MS/MS ?in a Q-Exactive mass spectrometry. The sequence of the representative ubiquitinated ?peptide is indicated?and the fragment ions are labeled in the MS/MS ?spectrum.? The diglycine-modified lysine and its position in the peptide ?sequence are indicated?with “gl” above?'K'.

Scientific Description

Ubiquitin(Ub) is a highly conserved protein of 8 kDa that covalently attaches to lysine residues of target proteins. This occurs through a three-step process involving Ub-activating (E1), Ub-conjugating (E2) and Ub-ligating (E3) enzymes. Ubiquitin-dependent processes are commonly modulated via formation of polyubiquitin chains, whereby the C terminus of a chain extending ubiquitin becomes linked to the N terminus or one of seven lysine residues (Lys-6,Lys-11, Lys-27, Lys-29, Lys-33, Lys-48, and Lys-63) within a substrate-bound ubiquitin molecule. The modification of lysine ubiquitin (ubiquitination) plays a central role in the regulation of multiple cellular processes. In addition to targeting proteins for proteasomal degradation, ubiquitination modulates membrane protein trafficking, alters protein-protein interactions, and controls the activity of many signal transduction pathways. A large family of enzymes involved in ubiquitin conjugation controls these diverse functions.

Storage & Stability

Store product at -20oC. Avoid repeated freeze/thaw. Antibody is supplied in PBS with 50% glycerol and 0.01% sodium azide.Stable for 12 months from date of receipt.

**Research purposes only. Not intended to be used for therapeutic or diagnostic purposes in humans or animals.

1.?Guoqiang,?X.,?et?al.?(2010).?Nat?Biotech.?28:868-873;

2.?Daisy,?B.,?et?al.?(2012).?Mol.Cell.Pro.?2012,?11:?1529-1540.

IAC ?IAC=Immunoaffinity Immunoprecipitation