Cells were lysed in IPH buffer (50 mM Tris-Cl, 0.5% NP-40%, 50 mM EDTA). Samples were run on a 10% polyacrylamide SDS-PAGE gel and transferred to nitrocellulose membranes. Membranes were blocked for 30 min in 5% milk containing PBST and incubated overnight with primary antibody in 1% milk. After washing, secondary antibody incubation was carried out for 1 h at 1:4000 dilution before washing and incubation with Millipore Immobilon HRP substrate. Antibodies used were as follows: MYOD1 sc-32758 (Santa Cruz Biotechnology), MHC 22287–1-AP (Proteintech), MYOGENIN sc-12732 (Santa Cruz Biotechnology), and HSP90 sc-13119 (Santa Cruz Biotechnology).
Myogenin sc 12732
Manufactured by Santa Cruz Biotechnology
Sourced in United States
MYOGENIN sc-12732 is a laboratory antibody product. It is an antibody that specifically binds to the myogenin protein, which is a transcription factor involved in muscle cell differentiation. The antibody can be used for the detection and analysis of myogenin in various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Hsp90 sc 13119, by Santa Cruz Biotechnology (3 mentions)
Mhc 22287 1 ap, by Proteintech (3 mentions)
Myod1 sc 32758, by Santa Cruz Biotechnology (3 mentions)
Immobilon hrp substrate, by Merck Group (2 mentions)
Myhc mf20, by Developmental Studies Hybridoma Bank (1 mentions)
Ripa bufer, by Thermo Fisher Scientific (1 mentions)
Erk1 2 4695, by Cell Signaling Technology (1 mentions)
Phospho erk1 2 4376, by Cell Signaling Technology (1 mentions)
Cav1 610059, by BD (1 mentions)
Protease inhibitor cocktail tablet, by Roche (1 mentions)
Phostop, by Roche (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Ab15277, by Abcam (1 mentions)
Ps9 gsk3β, by Cell Signaling Technology (1 mentions)
Ab11575, by Abcam (1 mentions)
P sapk jnk, by Cell Signaling Technology (1 mentions)
Sapk jnk, by Cell Signaling Technology (1 mentions)
Mab1675, by R&D Systems (1 mentions)
Vinculin v9131, by Merck Group (1 mentions)
Total paxillin, by BD (1 mentions)
6 protocols using myogenin sc 12732
1
Protein Expression Analysis in Myoblasts
2
Western Blot Analysis of Protein Expression
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Cells were lysed with radioimmunoprecipitation assay buffer (RIPA Bufer, Thermo Scientific) containing protease inhibitors (Complete, Mini; Protease Inhibitor Cocktail Tablets, Roche) and phosphatase inhibitors (PhoStop, Phospatase Inhibitor Cocktail Tablets, Roche) and centrifuged at 13000×g, at 4ºC, for 20 minutes. The protein content of the supernatants was determined with BCA assay system (Pierce). Lysate aliquots (50 μg) were resolved by 8%, 10% or 12% (depending on the protein molecular weight) SDS-PAGE and transferred onto nitrocellulose membranes. After blocking with 5% skimmed milk in PBS containing 0.2% Tween 20 at room temperature for 1 hour, membranes were incubated overnight at 4ºC with the appropriate primary antibody (CAV1 #610059 from BD, FoxO1 #2880, ERK1/2 #4695, phospho-ERK1/2 #4376 from Cell Signaling Technology; CAV3 #sc5310 and Myogenin #sc-12732 from Santa Cruz; MyHC MF 20 from Developmental Studies Hybridoma Bank). Blots were then incubated at room temperature for 1 hour with a horseradish peroxidase–conjugated secondary antibody and the peroxidase activity was detected by enhanced chemiluminescence (Pierce) following the instructions of the manufacturer. Immunodetection of β-actin (#ab49900) from Abcam was used as a loading reference.
3
Protein Extraction and Western Blot Analysis
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Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl [pH 7.4], 150 mM NaCl, 1% Triton X-100, 1 mM ethylenediaminetetraacetic acid, 1 mM ethylene glycol tetraacetic acid, 0.1% sodium dodecyl sulfate [SDS], 1% sodium deoxycholate, 1 mM Na3VO4, 1 mM sodium fluoride, 1 mM phenylmethylsulfonyl fluoride, and protease inhibitor cocktail). After a 30-minute incubation on ice, the lysates were centrifuged at 14,000 rpm for 20 minutes at 4°C. The protein concentration was measured with a bicinchoninic Acid (BCA) protein assay kit (Pierce Chemical Co., Rockford, IL, USA). Protein samples were separated by SDS-polyacrylamide gel electrophoresis (PAGE), transferred to a polyvinylidene fluoride membrane, and immunoblotted with antibodies [25 (link)]. The primary antibodies were: MyHC (MF20); myogenin (sc-12732) and CCR5 (sc-178933) (Santa Cruz Biotechnology, Dallas, TX, USA); and β-tubulin (T2200; Sigma-Aldrich).
4
Rabbit Polyclonal Antibodies for SLK
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Our rabbit custom polyclonal antibodies for SLK was used throughout this study, as previously described [35 (link)]. GAPDH (#5174), P-Y397-FAK (#3283), P-Y118-Paxillin (#2541), P-p38 (#4511), p38 (#9212), P-S9-GSK3β (#9336), GSK3β (#9315), P-SAPK/JNK (#4668), and SAPK/JNK(#9252), were acquired from cell signaling. MHC type 1/slow (M8421), MHC type 2/fast (M4276), γ tubulin (T5192) and Vinculin (V9131) were purchased from Sigma-Aldrich. Antibodies for dystrophin (ab15277) and laminin (ab11575) from Abcam were used for staining. The Myf5 (sc-302) and Myogenin (sc-12732) antibodies were purchased from Santa Cruz. Antibodies for periostin (AF2955) and Pax7 (MAB1675) were from R&D Systems. Finally, total FAK (610088) and total paxillin (612405) were acquired from BD Biosciences.
5
Protein Expression Analysis in Myoblasts
6
Western Blot Analysis of Myogenic Markers
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Cells were lysed in IPH buffer (50 mM Tris-Cl, 0.5% NP-40%, 50 mM EDTA). Samples were run on a 10% polyacrylamide SDS-PAGE gel and transferred to nitrocellulose membranes. Membranes were blocked for 30 min in 5% milk containing PBST and incubated overnight with primary antibody in 1% milk. After washing, secondary antibody incubation was carried out for 1 h at 1:4000 dilution before washing and incubation with Millipore Immonilon HRP substrate. Antibodies used were as follows: MYOD1 sc-32758 (Santa Cruz Biotechnology), MHC 22287-1-AP (Proteintech), MYOGENIN sc-12732 (Santa Cruz Biotechnology), and HSP90 sc-13119 (Santa Cruz Biotechnology).
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