Protein was extracted with RIPA buffer (Thermo Fisher) supplemented with protease inhibitor cocktails I and II (Sigma) and phosphatase inhibitor cocktail set III (Calbiochem). Protein was quantified with Pierce BCA protein assay kit (Thermo Fisher) and analysed with a Sunrise Tecan plate reader. After Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and electro‐transfer onto Polyvinyl‐Difluor membranes (GE Healthcare), Western blotting was performed with the following primary antibodies and concentrations: GAPDH (sc‐365062; Santa Cruz; 1:10,000), PAX3 (AB_528426; DSHB; 0.5 μg/mL), Twist2 (ab66031; Abcam; 1 μg/mL), IMP1/2/3 (sc‐271785; Santa Cruz; 1:1000), HMGA2 (no. 5269S; Cell Signalling; 1:1000), tubulin (ab210797; Abcam; 1:1000), p53 (sc‐126; Santa Cruz; 1:100), citrate synthase (G‐3; sc‐390693; Santa Cruz; 1:1000). Blots were stained with secondary antibodies conjugated with Horseradish Peroxidase, 1:2500, (Promega W401B, W402B) and visualized with ECL prime western blotting detection reagent kit (GE Healthcare).
Imp1 2 3
Manufactured by Santa Cruz Biotechnology
IMP1/2/3 are recombinant proteins produced by Santa Cruz Biotechnology. They are involved in the regulation of gene expression and mRNA stability. IMP1, IMP2, and IMP3 belong to the IGF2BP (Insulin-like Growth Factor 2 mRNA-Binding Protein) family and function as RNA-binding proteins.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
Protease inhibitor cocktails 1 and 2, by Merck Group (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Hmga2, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase, by Promega (1 mentions)
Twist2, by Abcam (1 mentions)
Ecl prime western blotting detection reagent kit, by GE Healthcare (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Igf2bp2, by Proteintech (1 mentions)
Sc 390693, by Santa Cruz Biotechnology (1 mentions)
2 protocols using imp1 2 3
1
Comprehensive Protein Extraction and Western Blot Analysis
2
Protein Extraction and Western Blotting
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Protein was extracted with RIPA buffer supplemented with protease inhibitor cocktails I and II (Sigma) and phosphatase inhibitor cocktail set III (Calbiochem). Protein was quantified with Pierce BCA protein assay kit (Thermo Fisher) and analysed with a Sunrise Tecan plate reader. After SDS-PAGE and electro-transfer onto PVDF membranes, Western blotting was performed with the following primary antibodies and concentrations: Lin28a (1:1000, CST), Pax7 (0.28μg/ml, DSHB), Pax3 (0.31μg/ml, DSHB), MyoD (1:1000, Santa Cruz Biotechnology), MHC (0.23μg/ml, DSHB), MyoG (1:1000, Santa Cruz Biotechnology), IGF2BP2 (1:1000, Proteintech), Hmga2 (1:1000, CST), Cre (1:1000, Millipore), GAPDH (1:1000, CST), Vinculin (K106900P, 1:5000, Solarbio), Tubulin (ab210797; Abcam; 1:1000), P53 (sc-126; Santa Cruz; 1:100), IMP1/2/3 (sc-271785; Santa Cruz; 1:1000), citrate synthase (G-3) (sc-390693; Santa Cruz ; 1:1000).
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