DNA transfection Mammalian cells Primary cells

Acid phosphatase detection heavily relies on determining the concentration of tartrate-resistant acid phosphatase (TRAP) in the sample. Hence, sample preparation is very crucial and it should be done strictly as per kit manufacturer instructions to avoid any inconsistency and poor sensitivity

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 Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 AT2R

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 AT1R

Get tips on using Expi293™ Expression System Kit to perform Protein expression and purification Mammalian cells - HEK 293 ECD

Get tips on using T-PER™ Tissue Protein Extraction Reagent to perform Protein isolation Mammalian cells - Mouse Epididymal fat

Get tips on using Pierce™ Coomassie (Bradford) Protein Assay Kit to perform Protein quantification Mammalian cells - MDA-MB-231

Get tips on using Mem-PER™ Plus Membrane Protein Extraction Kit to perform Protein isolation Mammalian cells - Mouse_Brown fat

Get tips on using PRO-PREP™ Protein Extraction Solution (C/T) to perform Protein isolation Mammalian cells - 3T3-L1

Get tips on using VWR Life Science RIPA Lysis Buffer, Biotechnology Grade to perform Protein isolation Mammalian cells - Caco-2