Transfected cells were lysed in lysis buffer (50 mM Tris–HCl, 150 mM NaCl, 2 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate) containing RNase inhibitors and protease inhibitors (Thermo Scientific, Waltham, MA, USA). The extracted protein was incubated with relevant antibodies (anti-ELAVL1 and IgG as control) (Abcam, USA) overnight at 4 °C and then pulled down with protein G Sepharose 4 Fast Flow suspension (GE Amersham, Little Chalfont, UK). The beads were digested with proteinase K (Sangon, Shanghai, China) for 1 h followed by RNA purification with Trizol reagent (Invitrogen, Missouri, USA). qRT-PCR was performed to examine the RNA yield. The primers were listed in the qRT-PCR section.
Rnase inhibitors and protease inhibitors
Manufactured by Thermo Fisher Scientific
Sourced in United States
RNase inhibitors and protease inhibitors are lab equipment products used to prevent the degradation of RNA and proteins, respectively, during experimental procedures. They work by inhibiting the activity of RNase and protease enzymes, which can break down RNA and proteins. These products help maintain the integrity of samples and ensure reliable experimental results.
Automatically generated - may contain errors
2 protocols using rnase inhibitors and protease inhibitors
1
RNA Binding Protein Immunoprecipitation
2
Chromatin Immunoprecipitation and RNA Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Transfected cells were lysed in lysis buffer (50 mM Tris–HCl, 150 mM NaCl, 2 mM EDTA, 1% NP-40, 0.5% sodium deoxycholate) containing RNase inhibitors and protease inhibitors (Thermo Scientific, Waltham, MA, USA). The extracted protein was incubated with relevant antibodies (anti-EZH2, 1:1 000 dilution and IgG as control) (Abcam) overnight at 4 °C and then pulled down with protein G Sepharose 4 Fast Flow suspension (GE Amersham, Little Chalfont, UK). The beads were digested with proteinase K (Sangon, Shanghai, China) for 1 h followed by RNA purification with Trizol reagent (Invitrogen). Quantitative RT-PCR was performed to examine the RNA yield.
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