Protein Expression Eukaryotic cells CHO

In ChIP, the most vital step is the binding of an antibody and choosing the right antibody. The binding affinity of different types of immunoglobulins to protein A or G differs significantly. Henceforth, it is recommended to choose either protein A or protein G coated beads. If you do not see any product in the positive control, add 5–10 μg of chromatin and 1–5 μg of antibody to each IP reaction and incubate with antibody overnight and an additional 2 hr after adding Protein G/A beads. If no product in the experimental, add more DNA to the PCR reaction or increase the number of amplification cycles. Choose an alternate, ChIP-validated antibody if the antibody does not work.

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - Caco-2

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - SH-SY5Y

Get tips on using CelLytic™ M to perform Protein isolation Mammalian cells - BHK-21

When extracting nucleic acids from cell cultures, thorough homogenization of cells via vortexing in lysis buffer is very necessary. Choose the best RNA isolation method keeping in mind the downstream applications, generally, column-based isolations result in clean and concentrated RNA samples. Downstream applications like sequencing and cDNA synthesis require high-quality RNA, always treat the samples with DNases and check their integrity by running a gel.

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - HepG2

Get tips on using Mammalian Cell Lysis kit to perform Protein isolation Mammalian cells - STTG1

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Human lung fibroblasts

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Mouse Epididymal fat