siRNA / RNAi /miRNA transfection Human Cells THP-1

Get tips on using GenJet™ In Vitro DNA Transfection Reagent to perform DNA transfection Mammalian cells - Primary cells Human lung fibroblasts (HLF)

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Cells - immortalized THP-1

Get tips on using Human HO 1 ELISA Kit (ab133064) to perform ELISA Human - HO-1

Get tips on using Human Dkk-1 ELISA Kit (RAB0143) to perform ELISA Human - Dkk-1

Get tips on using Human Dkk-1 DuoSet ELISA (DY1906) to perform ELISA Human - Dkk-1

Get tips on using Human Syndecan-1 DuoSet ELISA to perform ELISA Human - SDC1

DNA-protein interactions are studied by using ChIP. The basic steps in this technique are crosslinking, sonication, immunoprecipitation, and analysis of the immunoprecipitated DNA. During ChIP, if chromatin is under-fragmented or fragments are too large which can lead to the increased background and lower resolution. Shorter cross-linking times (5-10 min) and/or lower formaldehyde concentrations (<1%) may improve shearing efficiency. If Chromatin is over-fragmented, then optimize shearing conditions for each cell type to improve ChIP efficiency. Over-sonication of chromatin may disrupt chromatin integrity and denature antibody epitopes. If you do not see any product or very little product in the input PCR reactions, add 5–10 μg chromatin per IP.

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using Cell Lysis Buffer (10X) to perform Protein isolation Mammalian cells - THP-1

Get tips on using MethylFlash Methylated DNA 5-mC Quantification Kit to perform DNA quantification Human - THP 1