Total liver and muscle lysates were prepared in RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EGTA, 1% NP-40, 1% sodium deoxycholate, 1 mM Na3VO4) supplemented with protease and phosphatase inhibitor cocktails (Complete and PhosSTOP tablets, Roche). Membranes were probed with indicated antibodies (see resource table for ref.).
Complete and phosstop tablets
Manufactured by Roche
Sourced in Switzerland
Complete and PhosSTOP tablets are laboratory reagents designed for use in Western blot analysis. Complete tablets contain a mixture of protease inhibitors to prevent protein degradation, while PhosSTOP tablets contain a combination of phosphatase inhibitors to preserve protein phosphorylation states. Both products are intended to be used in sample preparation for Western blot experiments.
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Lab products found in correlation
5 protocols using complete and phosstop tablets
1
Liver and Muscle Lysate Preparation
2
Liver Protein Extraction and Western Blot
3
Liver and Muscle Lysate Preparation
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Total liver and muscle lysates were prepared in RIPA buffer (20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM EGTA, 1% NP-40, 1% sodium deoxycholate, 1 mM Na3VO4) supplemented with protease and phosphatase inhibitor cocktails (Complete and PhosSTOP tablets, Roche). Membranes were probed with indicated antibodies (see resource table for ref.).
4
Ex-Pt#4 Tumor Tissue Culture
5
Tumor Tissue Collection and Culture
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Tumors were collected following informed patient consent at Thomas Jefferson University Hospital under an IRB-approved protocol (#07D.483). The following PTC samples were isolated: TJU-THY #1 is from a 53 y/o, TJU-THY#2 is from a 32 y/o female, and TJU-THY#3 is from a 38 y/o. Less than 16 h post-surgery, excess adipose and stromal tissue was removed and tumors were cut into 1 mm3 pieces. Vetspon absorbable hemostatic gelatin 1 cm3 sponges (Novartis; Basel, Switzerland) were pre-soaked in 12-welled plates for 15 min at 37 °C in 500 μL of DMEM/10% FBS containing drugs or DMSO as a vehicle control. To avoid concerns of intratumoral heterogeneity, up to three 1 mm3 pieces from different locations of the original tumor were placed per sponge per treatment condition. Medium was replaced every 24 h. Tumor pieces for western blotting were homogenized in modified RPPA lysis buffer [75 (link)] with phosphatase and inhibitors (PhosSTOP and cOmplete tablets Roche, Basel, Switzerland). Laemmli sample buffer was added, and samples were heated at ≥95 °C for 5 min. In parallel, the lysates were sent to MD Anderson and used for reverse phase protein array (RPPA) analysis as previously described [46 (link)]. Data was visualized using Morpheus (Broad Institute).
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