Immunoprecision assays and mass spectrometry for Annexin A1 and PDZ and LIM domain 7 in stochastic vacuum fractions
| Title | Immunoprecision assays and mass spectrometry for Annexin A1 and PDZ and LIM domain 7 in stochastic vacuum fractions |
|---|---|
| Description | For immunoprecipitation (IP) assays, protein lysates were prepared from stochastic vacuum fractions (SVFs) from white adipose tissue of mice, using Pierce TM IP Lysis Buffer (Thermo, 87787). The lysates were incubated overnight at 4 centigrade with anti-ANXA1(Abcam ab214486, 1:100), anti-PDLIM7 (Proteintech 10221-1-AP, 1:100), or control immunoglobulin G (IgG; Beyotime, A7058) on a rotating pattern. Immobilized Protein A/G resin slurry (Thermo, 20423) was added to the lysates and incubated for 2 hours at 4 centigrade with gentle mixing. The complexes were washed five times with lysis buffer (Thermo, 28379) and resuspended in 2-SDS loading buffer. The immunoprecipitated proteins were eluted by incubation at 95°C for 5 minutes and detected by immunoblotting after separation by SDS-PAGE.For mass spectrometry-based proteomic analysis, gel pieces were dehydrated with acetonitrile and digested with trypsin. LC-MS/MS analysis of peptides was conducted at Peking University Institute of Systems Biomedicine (China). |
| Organism | Mus musculus |
| Data Type | Proteomic Data by Mass Spectrometry (MS) |
| Data Accessibility | Open-access |
| BioProject | PRJCA027679 |
| Release Date | 2024-08-02 |
| Submitter | Lu Fang (fanglu17@pku.edu.cn) |
| Organization | Peking University |
| Submission Date | 2024-07-03 |
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| File ID | File Title | Number/Samples | File Type | File Size | File Suffix | Download Times | Download |
|---|---|---|---|---|---|---|---|
| OMIX006798-01 | IP-MS(ANXA1-IgG) | 1 | Proteomic Data by Mass Spectrometry (MS) | 556.1 KB | xlsx | 0 | |
| OMIX006798-02 | IP-MS(PDLIM7-IgG) | 1 | Proteomic Data by Mass Spectrometry (MS) | 128.5 KB | xlsx | 0 |