siRNA / miRNA gene silencing Human Melanoma cells (501 Mel and SK Mel 28)

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Get tips on using GeneChip® HT 3' IVT PLUS Reagent Kit to perform Microarray RNA amplification & Labeling - Human endometrial stromal cells Biotin

Get tips on using Freedom™ DG44 Kit to perform Protein expression and purification Mammalian cells - CHO-K1 G12

Get tips on using DuoSet® ELISA Development Systems to perform ELISA (kit) IL-6, IL-1β, IL-8 and TNF-α - -NA- Human -NA-

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.

Get tips on using In Vitro Angiogenesis Assay Kit to perform Angiogenesis assay human - bone marrow mononuclear cells

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human dermal fibroblasts

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human osteoblasts - osteoarthritis

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human lung fibroblasts

Get tips on using TRIzol Reagent to perform RNA isolation / purification Cells - primary human epidermal keratinocytes