Using the ChIP Assay Kit (Boytime, Nanjing, China) following the manufacturer's instruction to detect ChIP-PCR. In brief, using 1% formaldehyde to cross-link porcine pre-adipocyte at 37℃ for 10 minutes. 1×glycine is used to quench the cross-linked porcine pre-adipocyte at room temperature for 5 minutes. Using VCX750, the ultrasonic cell smash machine (Sonics, United States) to smash porcine pre-adipocyte and obtain DNA fragments. Subsequently, following the rule that per 100ml cell lysis buffer is incubating with 1 mg of anti-C/EBPα (ZEN BIO, 383901) and anti-FoxO1 (ZEN BIO, 383312) at 4℃ overnight, 60ml Protein A Agarose/SalmonSperm DNA is used to isolate immunoprecipitated complexes at 4℃ for 4 hours, washing the complexes as following: low salt wash buffer, high salt wash buffer, LiCl wash buffer once, and TE buffer twice. Final ChIP DNA fragments are subjected to PCR analysis using an AdipoQ promoter specific primers, as shown at Supplementary Table S1.
Ultrasonic cell smash machine
Manufactured by Sonics
Sourced in United States
The Ultrasonic cell smash machine is a laboratory equipment used for the disruption and lysis of cells, tissues, and other biological samples. It utilizes high-frequency sound waves to physically break down the cellular structure, allowing for the extraction of cellular contents.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using ultrasonic cell smash machine
1
ChIP Assay of Porcine Pre-adipocytes
2
ChIP Assay of Porcine Pre-adipocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Using the ChIP Assay Kit (Boytime, Nanjing, China) following the manufacturer's instruction to detect ChIP-PCR. In brief, using 1% formaldehyde to cross-link porcine pre-adipocyte at 37℃ for 10 minutes. 1×glycine is used to quench the cross-linked porcine pre-adipocyte at room temperature for 5 minutes. Using VCX750, the ultrasonic cell smash machine (Sonics, United States) to smash porcine pre-adipocyte and obtain DNA fragments. Subsequently, following the rule that per 100ml cell lysis buffer is incubating with 1 mg of anti-C/EBPα (ZEN BIO, 383901) and anti-FoxO1 (ZEN BIO, 383312) at 4℃ overnight, 60ml Protein A Agarose/SalmonSperm DNA is used to isolate immunoprecipitated complexes at 4℃ for 4 hours, washing the complexes as following: low salt wash buffer, high salt wash buffer, LiCl wash buffer once, and TE buffer twice. Final ChIP DNA fragments are subjected to PCR analysis using an AdipoQ promoter specific primers, as shown at Supplementary Table S1.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!