Cells were lysed in RIPA buffer with protease inhibitors (Roche). After quantification using a BCA protein assay kit (Pierce), 40 µg of total protein was separated by 10% SDS-PAGE under denaturing conditions and transferred to PVDF membranes (GE Healthcare). Membranes were blocked in 5% BSA (Sigma) and then incubated with a Msi2 primary antibody (1:1000; Novus Biologicals), Msi1 (1:1,000; MBL), NICD (1:1,000; Abcam), Hes1 (1:1,000; Abcam), β-actin (1:10000, Abcam), or Pten (1:1000, Cell Signaling) APC (1:200, Santa Cruz sc-896) followed by incubation with a secondary antibody conjugated with horseradish peroxidase (HRP) (1:2,000; Cell Signaling) together with an HRP-conjugated primary antibody for β-actin (1:10,000; Sigma). Immunoreactive proteins were visualized using LumiGLO chemiluminescent substrate (Pierce). Raw scans of Western blots in Figures 4C , 5C , 6F , and 7A can be found in the supplementary information accompanying the manuscript.
Hrp conjugated primary antibody for β actin
Manufactured by Merck Group
The HRP-conjugated primary antibody for β-actin is a laboratory reagent used for the detection and quantification of the β-actin protein in various biological samples. It consists of a primary antibody specific to the β-actin protein, which is conjugated to the enzyme horseradish peroxidase (HRP). The HRP-conjugation allows for the amplification of the signal during immunodetection procedures, facilitating the visualization and quantification of the target protein.
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2 protocols using hrp conjugated primary antibody for β actin
1
Western Blot Analysis of Protein Expression
2
Western Blot Analysis of Protein Expression
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Cells were lysed in RIPA buffer with protease inhibitors (Roche). After quantification using a BCA protein assay kit (Pierce), 40 µg of total protein was separated by 10% SDS-PAGE under denaturing conditions and transferred to PVDF membranes (GE Healthcare). Membranes were blocked in 5% BSA (Sigma) and then incubated with a Msi2 primary antibody (1:1000; Novus Biologicals), Msi1 (1:1,000; MBL), NICD (1:1,000; Abcam), Hes1 (1:1,000; Abcam), β-actin (1:10000, Abcam), or Pten (1:1000, Cell Signaling) APC (1:200, Santa Cruz sc-896) followed by incubation with a secondary antibody conjugated with horseradish peroxidase (HRP) (1:2,000; Cell Signaling) together with an HRP-conjugated primary antibody for β-actin (1:10,000; Sigma). Immunoreactive proteins were visualized using LumiGLO chemiluminescent substrate (Pierce). Raw scans of Western blots in Figures 4C , 5C , 6F , and 7A can be found in the supplementary information accompanying the manuscript.
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