NanoLC-MS/MS

LC-MS/MS including GeLC-MS/MS Proteomics Analysis can identify all proteins in a mixture ranging from 1D gel bands, IP samples to whole cell lysate or tissues. For Moderate to high complexity samples, customers may consider the following additional options to improve sensitivity:

  1. Protein fractionations: Fractionate proteins by either pI or molecular weight at your choice. This is very useful for tissues, cell lysates, or culture media.
  2. Depletion of serum abundant proteins: Please note that LC-MS/MS detection of serum low abundant proteins is limited even after depletion of serum abundant proteins. We highly recommend Serum 2D DIGE for all serum and plasma samples.

Sample category:

  1. Low complexity samples: 1D gel band – contain only one gel piece with one protein band, no bigger than 1x1x5mm in size, <10 proteins
  2. Moderate complexity samples: protein mixture such as column purified sample, <100 proteins
  3. Moderate to High complexity samples: Protein mixtures such as IP samples, mitochondria, endoplasmic reticulum, lysosomes and other subcellular organelles
  4. High complexity samples: whole cell lysate, tissue, and others with more than 500 protein mix

nanoLC-MS/MS Procedure

LCMSMS procedure: step 1 - in gel or in solution samples

Protein Mixture (1D Gel bands or Liquid samples)

LCMSMS procedure: step 2 - Protein Fractionation by pI or Mw

Protein Fractionation by pI or Mw (optional)

LCMSMS procedure: step 3 - Reduction/alkylation & Trypsin Digestion

Reduction/alkylation & Trypsin Digestion

LCMSMS procedure: step 4 - peptide separation by Nano-HPLC

Nano-HPLC

LCMSMS procedure: step 5 - prptide fragmentation by MSMS

MS/MS

LCMSMS procedure: step 6 - Database Search for Protein ID

Database Search for Protein ID

In-solution Samples

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

Protein amount: 5-200 µg of total protein from each sample.

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 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 view here for the rank of biohazardous materials.

Sample type: Please inform our scientists about your sample type. We will send you additional tips for each sample type.

Pricing

Services DescriptionAcademic/Governmental LabsIndustrial/Commercial Labs
CodePriceCodePrice
Protein Fractionation355Acall for price355Ncall for price
LC-MS/MS on Low Complexity Samples
(1D Gel band, 2-5 samples)
331A$450/sample331N$540/sample
LC-MS/MS on Low Complexity Samples
(1D Gel band, 6 or more samples)
331A$380/sample331N$450/sample
LC-MS/MS on Moderate Complexity Samples332Acall for price332Ncall for price
LC-MS/MS on Moderate to High Complexity Samples334Acall for price334Ncall for price
LC-MS/MS on High Complexity Samples333Acall for price333Ncall for price

Each order should have a minimum of 2 samples. Turnaround time is ~7 business days.

Sample category:

  • Low complexity samples: 1D gel band – Each sample should contain only one gel piece with one protein band, no bigger than 1x1x5mm in size, <10 proteins. Samples with more than one gel piece, and/or more than one protein band will be considered as moderate to high complex samples. We recommend you cut out only the area with your protein of interest (1x1x2mm size) and follow these guidelines to prepare samples
  • Moderate complexity samples: protein mixture such as column purified sample, <100 proteins 
  • Moderate to High complexity samples: Protein mixtures such as subcellular organelles, IP samples with 100 – 500 proteins
  • High complexity samples: whole cell lysate, tissue, and others with more than 500 protein mix

Price covers the following:

  • Nano-LC coupled with auto-spotting
  • MS & MS/MS
  • Standard database search against one species in NCBI or SwissProt database using MASCOT
  • Data report (see an example below)

Price does NOT cover:

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