Immunohistochemistry Anti-Glial Fibrillary Acidic Protein (GFAP) Mouse

Get tips on using β-Actin Antibody (C4): sc-47778 to perform Western blotting β-Actin

Get tips on using Pre-designed and validated siRNA against gene IGFBP1 to perform siRNA / miRNA gene silencing Human - Primary Endometrial Stromal Cells IGFBP1 (Insuline-like growth factor binding protein-1)

Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - Bone marrow-derived macrophages (BMDMs)

Get tips on using Active/Pro-Caspase 3 Monoclonal Antibody (31A1067) to perform Western blotting Caspase-3

Get tips on using ShRNA CD24 Lentiviral Transduction Particles (CD24-V2LHS_71909)) to perform shRNA gene silencing Mouse - Prostate cancer cell lines (DU145 and PC3) CD24 lentiviral particles

Get tips on using ShRNA CD24 Lentiviral Transduction Particles (CD24-V2LHS_71908) to perform shRNA gene silencing Mouse - Prostate cancer cell lines (DU145 and PC3) CD24 lentiviral particles

Get tips on using BLOCK-iT™ Adenoviral RNAi Expression System, pAd/BLOCK-iT™-DEST RNAi Gateway Vector to perform shRNA gene silencing Mouse - P19 Foxm1

Get tips on using Muse™Autophagy LC3-antibody based Kit to perform Autophagy assay cell type - 143B

Get tips on using MouseTRAP™ (TRAcP 5b) ELISA to perform Acid phosphatase assay cell type - murine macrophage cells

Plasmid isolation is an important technique in molecular biology or any kind of genetic editing. It involves amplifying plasmids overnight by transforming them into competent bacterial cells. The desired colonies of these bacteria can then be grown in shaker cultures, at appropriate shaking speed, oxygen availability and temperature. These liquid cultures can then be ultracentrifuged to pellet the bacteria, which are then used for plasmid isolation. The bacteria are first resuspended in a buffer, then lysed, neutralized, purified in a column, eluted, precipitated with ethanol and then resuspended. During plasmid isolation, it is important to lyse cells quickly because lysing bacteria for too long may lead to irreversible denaturing of the plasmid. Usually, alkaline lysis is used for isolation because it is a mild treatment. It isolates plasmid DNA and other cell components such as proteins by breaking cells apart with an alkaline solution. Precipitation removes the proteins, and the plasmid DNA recovers with alcohol precipitation. Resuspension and lysis buffers should be mixed thoroughly in order to prevent the DNA from breaking into smaller fragments. This is because broken gDNA can reanneal and remain in the solution, without binding to the column.