Prostate cancer cell lines LAPC4 and LNCaP were purchased from American Type Culture Collection (ATCC, Manassas, VA). RPMI media, SILAC RPMI media, fetal bovine serum (FBS), dialyzed fetal bovine serum (dFBS), Geneticin (G418), puromycin, 100 U/mL penicillin/streptomycin were from Life Technologies (Grand Island, NY). SILAC isotopes: 13C614N4-l -lysine (k 6) and 13C615N4-L-arginine (R10) were from Cambridge Isotope Laboratories, Inc. (Tewksbury, MA). OptiPrep was from Sigma-Aldrich (St. Louis, MO). Mouse anti-ALIX (1A12) was from Santa Cruz Biotechnology (Dallas, TX), and the rabbit anti-FUT8 was a kind gift from Drs. Ruey-Bing Yang and Chen-Fen Tu (Institute of Biomedical Sciences, Academia Sinica, Taipei).28 (link) NuPAGE precast gels and NuOAFE MES running buffer were from Thermo Fisher Scientific (Waltham, MA). Trypsin was from Promega (Madison, WI), and lysyl endopeptidase was from Wako Laboratory Chemicals USA (Richmond, VA). BCA assay kit Pierce was from Thermo Fisher Scientific (Rockford, IL). C18 SepPak columns and Oasis MAX extraction columns were purchased from Waters Corporation (Milford, MA). C18 solid-phase extraction disks were from 3M (St. Paul, MN).
Protocol detail
Quantitative Proteomic Analysis of Prostate Cancer
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LAPC4
LNCaP
RPMI media
fetal bovine serum (FBS),
penicillin/streptomycin
OptiPrep
Mouse anti-ALIX (1A12)
NuPAGE precast gels
Trypsin
C18 SepPak columns
Arginine
Assay
Buffer
Cell lines
Fetal bovine serum
G418
Gels
Geneticin
Isotope
Laboratory chemicals
Lysine
Lysyl endopeptidase
Mouse
Oasis
Penicillin
Promega
Prostate cancer
Puromycin
Rabbit
Solid phase extraction
Streptomycin
Trypsin
Corresponding Organization : Johns Hopkins University
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Variable analysis
independent variables
- LAPC4 and LNCaP prostate cancer cell lines
- Not explicitly mentioned
- RPMI media
- SILAC RPMI media
- Fetal bovine serum (FBS)
- Dialyzed fetal bovine serum (dFBS)
- Geneticin (G418)
- Puromycin
- 100 U/mL penicillin/streptomycin
- SILAC isotopes: 13C6 14N4-L-lysine (K6) and 13C6 15N4-L-arginine (R10)
- OptiPrep
- NuPAGE precast gels and NuOAFE MES running buffer
- Trypsin
- Lysyl endopeptidase
- BCA assay kit Pierce
- C18 SepPak columns and Oasis MAX extraction columns
- C18 solid-phase extraction disks
- Positive control: Mouse anti-ALIX (1A12) antibody from Santa Cruz Biotechnology
- Negative control: Rabbit anti-FUT8 antibody from Drs. Ruey-Bing Yang and Chen-Fen Tu (Institute of Biomedical Sciences, Academia Sinica, Taipei)
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