Proteins were obtained using whole cell lysis buffer (Roche, Basel, Switzerland) for 30 min on the ice. Protein samples were quantified using Pierce BCA Protein Assay kit (Thermo Fisher Scientific, Rockford, IL, USA). Then, 30 µg of proteins were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to the polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, USA). After blocking with 5% w/v nonfat dry milk for 1 h, primary antibodies were used to probe the indicated proteins followed by incubation with the secondary antibody (ab205718; Abcam, Cambridge, MA, USA). The protein bands were measured using the enhanced chemiluminescent (ECL) system (Beyotime, Shanghai, China) according to the manufacturer’s instructions. Gray analysis was conducted to quantify the expression of proteins using ImageJ software. Primary antibodies, including anti-BRD4 (ab128874), anti-phosphorylated-phosphatidylinositol 3-kinase (anti-p-PI3K; ab70912), anti-PI3K (ab32089), anti-p-AKT serine/threonine kinase (p-AKT; ab38449), anti-AKT (ab64148), anti-CD63 (ab59479), anti-CD81 (ab79559) and anti-β-actin (ab8226), were purchased from Abcam.
Whole cell lysis buffer
Manufactured by Roche
Sourced in Switzerland
Whole cell lysis buffer is a solution used to disrupt the cell membrane and release the contents of the cell, including proteins, nucleic acids, and other cellular components. This buffer facilitates the extraction and purification of these cellular components for further analysis or experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (4 mentions)
Ecl system, by Beyotime (4 mentions)
Ab38449, by Abcam (4 mentions)
Ab59479, by Abcam (4 mentions)
Ab70912, by Abcam (4 mentions)
Ab32089, by Abcam (4 mentions)
Ab79559, by Abcam (4 mentions)
Ab64148, by Abcam (4 mentions)
Ab128874, by Abcam (4 mentions)
Ab8226, by Abcam (4 mentions)
Ab205718, by Abcam (4 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
4 protocols using whole cell lysis buffer
1
Western Blot Analysis of Signaling Pathways
2
Western Blot Analysis of Protein Signaling
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Proteins were obtained using whole cell lysis buffer (Roche, Basel, Switzerland) for 30 min on the ice. Protein samples were quanti ed using Pierce BCA Protein Assay kit (Thermo Fisher Scienti c, Rockford, IL, USA). Then, 30 µg of proteins were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to the polyvinylidene uoride (PVDF) membrane (Millipore, Billerica, MA, USA). After blocking with 5% w/v nonfat dry milk for 1 h, primary antibodies were used to probe the indicated proteins followed by incubation with the secondary antibody (ab205718; Abcam, Cambridge, MA, USA). The protein bands were measured using the enhanced chemiluminescent (ECL) system (Beyotime, Shanghai, China) according to the manufacturer's instructions. Gray analysis was conducted to quantify the expression of proteins using ImageJ software. Primary antibodies, including anti-BRD4 (ab128874), anti-phosphorylated-phosphatidylinositol 3-kinase (anti-p-PI3K; ab70912), anti-PI3K (ab32089), anti-p-AKT serine/threonine kinase (p-AKT; ab38449), anti-AKT (ab64148), anti-CD63 (ab59479), anti-CD81 (ab79559) and anti-β-actin (ab8226), were purchased from Abcam.
3
Protein Expression and Quantification
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Proteins were obtained using whole cell lysis buffer (Roche, Basel, Switzerland) for 30 min on the ice.
Protein samples were quanti ed using Pierce BCA Protein Assay kit (Thermo Fisher Scienti c, Rockford, IL, USA). Then, 30 µg of proteins were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to the polyvinylidene uoride (PVDF) membrane (Millipore, Billerica, MA, USA). After blocking with 5% w/v nonfat dry milk for 1 h, primary antibodies were used to probe the indicated proteins followed by incubation with the secondary antibody (ab205718; Abcam, Cambridge, MA, USA). The protein bands were measured using the enhanced chemiluminescent (ECL) system (Beyotime, Shanghai, China) according to the manufacturer's instructions. Gray analysis was conducted to quantify the expression of proteins using ImageJ software. Primary antibodies, including anti-BRD4 (ab128874), anti-phosphorylated-phosphatidylinositol 3-kinase (anti-p-PI3K; ab70912), anti-PI3K (ab32089), anti-p-AKT serine/threonine kinase (p-AKT; ab38449), anti-AKT (ab64148), anti-CD63 (ab59479), anti-CD81 (ab79559) and anti-β-actin (ab8226), were purchased from Abcam.
Protein samples were quanti ed using Pierce BCA Protein Assay kit (Thermo Fisher Scienti c, Rockford, IL, USA). Then, 30 µg of proteins were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to the polyvinylidene uoride (PVDF) membrane (Millipore, Billerica, MA, USA). After blocking with 5% w/v nonfat dry milk for 1 h, primary antibodies were used to probe the indicated proteins followed by incubation with the secondary antibody (ab205718; Abcam, Cambridge, MA, USA). The protein bands were measured using the enhanced chemiluminescent (ECL) system (Beyotime, Shanghai, China) according to the manufacturer's instructions. Gray analysis was conducted to quantify the expression of proteins using ImageJ software. Primary antibodies, including anti-BRD4 (ab128874), anti-phosphorylated-phosphatidylinositol 3-kinase (anti-p-PI3K; ab70912), anti-PI3K (ab32089), anti-p-AKT serine/threonine kinase (p-AKT; ab38449), anti-AKT (ab64148), anti-CD63 (ab59479), anti-CD81 (ab79559) and anti-β-actin (ab8226), were purchased from Abcam.
4
Protein Expression Analysis via Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were obtained using whole cell lysis buffer (Roche, Basel, Switzerland). Proteins were run on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to the polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, USA). After blocking, primary antibodies were used to probe the indicated proteins followed by incubation with the secondary antibody (ab205718; Abcam, Cambridge, MA, USA). The protein bands were measured using the enhanced chemiluminescent (ECL) system (Beyotime, Shanghai, China). Gray analysis was conducted to quantify the expression of proteins. Primary antibodies, including anti-BRD4 (ab128874), anti-phosphorylated-phosphatidylinositol 3-kinase (anti-p-PI3K; ab70912), anti-PI3K (ab32089), anti-p-AKT serine/threonine kinase (p-AKT; ab38449), anti-AKT (ab64148), anti-CD63 (ab59479), anti-CD81 (ab79559) and anti-β-actin (ab8226), were purchased from Abcam.
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