LC/MS/MS Method Package for Mouse Progranulin

For LabSolutions Version 5 and LCMS-8080

Ready-to-Use Methods Provided

This Method Package allows for efficient MRM analysis using trypsin-digested progranulin without the need for the laborious tasks often required. Most users end these tasks, such as the selection of the target peptides, investigation of the analysis conditions, and optimization of the MS parameters with respect to the selected peptides, to be the most time-consuming part of method development.
By using this already developed method, the retention time is adjusted according to the HPLC configuration when the system is installed, and analysis can be started by simply providing the designated column, mobile phase, and reagents.

HPLC Conditions

Analytical column Shim-pack XR-ODSII
(2.0 mm I.D. × 150 mm L., 2.2 μm)
P/N 228-41623-93
Mobile phase A 5 mmol/L ammonium formate + 0.1 % Formic acid in water
Mobile phase B 0.1 % Formic acid in acetonitrile
Flow rate 0.3 mL/min
Injection volume 3 to 5 μL
Step Time(min) A(%) B(%)
0 0 97 3
1 15 60 40
2 5.50 60 40
3 5.51 97 3
4 8 97 3

Total Support from Pretreatment to Data Acquisition and Analysis

The method package includes an example of the sample preparation protocol for mouse serum, which can then be used as a guide for other biological samples. Since the sample preparation protocols can also be used for biological samples other than serum, this is a product that can be used by anyone engaged in analysis of various tissues cultures of mice. After data acquisition, quantitation can be performed using a calibration curve created from purified progranulin standard samples.

Quantitation of Individual Mature Proteins (Granulin)

The Method Package provides both simultaneous analysis methods and individual methods for trypsin-digested peptides at four positions to include the granulin-3 domain, granulin-4 domain, granulin-3 domain plus a linker domain, and lastly the granulin-4 domain plus a linker. The content of mature proteins after processing (granulin-3, granulin-4) can be calculated from the quantitation results for each peptide.

Progranulin and Granulin

Progranulin is a blood marker protein for insulin resistance that is characteristic of obesity and Type 2 diabetes. It is a hormone protein (adipokine) excreted by adipose tissue. Its elevated concentration in blood due to a high-fat diet induces insulin resistance. Granulin is approximately a 6 kDa active peptide that is excised from its progranulin precursor. Physiological response has been reported for mature granulin-3 and granulin-4. They are both expected to act as good detection markers for insulin resistance in much the same way as the progranulin precursor does.

Optimization through Q1 Scans and Product Ion Scans

Precursor ions and fragment ions are selected based on amino acid sequence information from Q1 scans and product ion scans using trypsin digests of progranulin.

Remarks and Precautions

  1. For Research Use Only. Not for use in diagnostic procedures.
  2. Shimadzu makes no warranty regarding the accuracy of information included in the database or the usefulness of information obtained from using the database.
  3. Be sure to perform tests using standard samples to confirm qualitative and quantitative information obtained with the given system.

For Research Use Only. Not for use in diagnostic procedures.

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