Identify Methylation Site

Each experiment covers identifying ALL Mono-Methylated Lysine Sites in a single protein per sample. This service works the best with purified proteins or IP samples with target proteins enriched. Customers can send samples in-gel or in-solution, and provide the amino acid sequence or accession number of the target protein.

The presence of a mono-methylated lysine residue is confirmed by the presence of an immonium ion peak at 98 m/z during MS/MS.

Methylation Site Identification Procedure

Gel Bands/Spots or In-Solution Samples

Procedure: step 1: cleaning and buffer exchange of in-gel or in-solution samples

Reduction/Alkylation & Trypsin Digestion

Procedure: step 2: Reduction, alkylation & trypsin digestion

MS

Peptide Without Methylated Lysine

Methylation Site Identification Procedure: step 3A: MS showing the Non-methylated peptide peak

Peptide With Methylated Lysine

Methylation Site Identification Procedure: step 3B: MS showing the methylated peptide peak

MS/MS

Peptide Without Methylated Lysine

Methylation Site Identification Procedure: step 4A: MSMS showing the absence of immonium ion peak at 126 m/z of Non-methylated peptide

Peptide With Methylated Lysine

Methylation Site Identification Procedure: step 4B: MSMS showing the presence of immonium ion peak at 126 m/z of methylated peptide

Database Search for Mono-Methylated Lysine Sites

Sample Info

Sample type: We accept in-solution, in-gel samples, or IP beads for this service.

Required information: Amino acid sequence or accession number of the target protein.

Protein amount: 2-20 µg of purified protein or protein mixture with target proteins enriched (such as IP).

Protein concentration: 5-20 mg/ml is preferred for in-solution samples.

Buffer: Please feel free to submit samples in whichever buffer that you would normally use.  Please inform our scientists about the buffer composition. Additional charge may apply on samples that require extensive amount of the extra work such as protein eluting, concentrating, buffer exchange or serum abundant protein depletion.

Biohazardous Materials: We do NOT accept samples containing any biohazardous materials. If your samples contain any biohazardous materials, please make sure to deactivate them completely using the appropriate procedure and declare them in the order form. In addition, we do NOT accept any samples containing Level 4 biohazardous materials even if they are deactivated. Please refer to this page for the rank of biohazardous materials.

Pricing

Services DescriptionAcademic/Governmental LabsIndustrial/Commercial Labs
CodePriceCodePrice
Identification of Mono-Methylation-site325A$769325N$899
Manual inspection of MS/MS spectrum202A$115202N$135

Each order should have a minimum of 2 samples. Turnaround time is 5-7 business days. Price covers the identification of all Mono-Methylation-site for 1 protein per sample; following are the major steps:

  • Reduction and alkylation
  • Trypsin digestion
  • Peptide extraction
  • Desalting
  • NanoLCMSMS on peptides matching the mass of methylated peptides from virtual digest
  • Standard database search against one species in NCBI or SwissProt database using MASCOT
  • Data report:
    1. Top 10 hits via MASCOT search
    2. List of precursors with predicated methylated sites
    3. MSMS spectrums on precursors showing an immonium ion peak at 98 m/z

Price does NOT cover:

  • Protein identification
  • Sample preparation
  • Data request in addition to the standard report

If a customer does not provide protein sequence or accession number, an additional $150 will be charged for protein identification.