Western blot analysis was performed with whole cell lysate. The cell monolayer was rinsed three times with ice-cold PBS prior to collecting cells with a cell lifter (VWR) and pelleting by centrifugation. The cell pellets were lysed in 20 mM HEPES pH8.0, 100 mM NaCl, 1 mM EDTA and 1% Tween20 containing protease inhibitor cocktail (ROCHE) on ice for 30 minutes, sonicated, and cell debris was cleared by centrifugation. The protein concentration of the supernatant was quantified using the micro BCA protein assay kit (Pierce). About 70–100 μg of total protein was separated on 4–12% SDS-PAGE gel (Invitrogen) and transferred to PVDF membranes (GE Healthcare) followed by incubation with anti-rabbit-DNMT3A antibody (Abcam ab188470 lot:GR224165–13) and anti-rabbit-GAPDH antibody (Cell Signaling 2118 lot: 10) as a loading control. All blots were developed using the ECL reaction kit (Bio-Rad) according to the manufacturer’s instructions.
Anti rabbit dnmt3a antibody
Manufactured by Abcam
Anti-rabbit-DNMT3A antibody is a laboratory tool used to detect and study the DNMT3A protein in various biological samples. It is a specific antibody raised against the DNMT3A protein in rabbits. The core function of this antibody is to bind to and identify the DNMT3A protein, enabling researchers to analyze its expression and localization within cells or tissues.
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Lab products found in correlation
2 protocols using anti rabbit dnmt3a antibody
1
Western Blot Analysis of DNMT3A Protein
2
Western Blot Analysis of DNMT3A Protein
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Western blot analysis was performed with whole cell lysate. The cell monolayer was rinsed three times with ice-cold PBS prior to collecting cells with a cell lifter (VWR) and pelleting by centrifugation. The cell pellets were lysed in 20 mM HEPES pH8.0, 100 mM NaCl, 1 mM EDTA and 1% Tween20 containing protease inhibitor cocktail (ROCHE) on ice for 30 minutes, sonicated, and cell debris was cleared by centrifugation. The protein concentration of the supernatant was quantified using the micro BCA protein assay kit (Pierce). About 70–100 μg of total protein was separated on 4–12% SDS-PAGE gel (Invitrogen) and transferred to PVDF membranes (GE Healthcare) followed by incubation with anti-rabbit-DNMT3A antibody (Abcam ab188470 lot:GR224165–13) and anti-rabbit-GAPDH antibody (Cell Signaling 2118 lot: 10) as a loading control. All blots were developed using the ECL reaction kit (Bio-Rad) according to the manufacturer’s instructions.
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