Identify Phosphorylation Site
Each experiment covers identifying ALL Phosphorylation sites (Ser, Thr and Tyr) 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.
Identification of Phosphorylation-site involves enriching phosphorylated peptides followed by mass spectrometry. The enrichment is critical since phosphorylated proteins often represent 1-2% of the total protein population. In each run, a positive control consisting of a peptide with a known phosphosite is subjected to the same experimental conditions as the test samples. The presence of phosphosites is confirmed at by the precense of a MSMS peak showing the loss of phosphate.
Phosphorylation Site Identification Procedure
Gel Bands/Spots or In-Solution Samples
Reduction/Alkylation & Trypsin Digestion
MS/MS: Phosphorylation Site
Database Search for Phosphorylation Sites
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.
|Services Description||Academic/Governmental Labs||Industrial/Commercial Labs|
|Identification of Phosphorylation-Site||322A||$695||322N||$810|
Each order should have a minimum of 2 samples. Turnaround time is 5-7 business days. Price covers the identification of phosphorylation sites for 1 protein per sample; following are the major steps:
- Reduction and alkylation
- Trypsin digestion
- Peptide extraction
- Phospho-peptide enrichment (if you order item # 321)
- NanoLCMSMS on peptides matching the mass of phosphophorylated peptides from virtual digest
- Standard database search against one species in NCBI or SwissProt database using MASCOT
- Data report:
- Top 10 hits via MASCOT search
- List of precursors with predicated phosphorylation site
- MSMS spectrums on precursors showing loss of phosphate
Price does NOT cover:
- Protein identification
- Sample preparation
- Data request in addition to the standard report
If customer does not provide protein sequence or accession number, an additional $150 will be charged for protein identification