siRNA / RNAi /miRNA transfection Rat AR42J

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 MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles to perform shRNA gene silencing Human - Islets of langerhans Negative control (scrambled) lentiviral particles

ELISA is the most commonly used method of detecting and quantifying the concentration of an antigen in an unknown sample. During the experiment, If you get a weak signal, then make sure reagents are at room temperature before starting the assay. Try increasing incubation times to ensure maximal antibody binding and amplify the signal. Secondly, if you get values above 0 in the negative control indicates a high background signal. Try to consider reducing your antibody concentration and prevent non-specific binding of antibodies by using affinity-purified antibody and suitable blocking buffers. To avoid high well to well variation, do not stack plates during incubation, no bubbles in the plate and wash wells thoroughly to avoid variation.

Reporter gene assays enable high sensitivity measurement of gene expression and cell signaling through the addition of bioluminescent genes into target cells. One of the major challenges is to make a specific construct that has no responses other than those related to the signaling pathway of interest. This can be achieved by selecting highly specific reporter constructs containing only defined responsive elements and a minimal promoter linked to reporter enzymes such as luciferase

Get tips on using Ras (D2C1) Rabbit mAb #8955 to perform Western blotting Ras

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Liver

Get tips on using RIPA Buffer to perform Protein isolation Mammalian cells - Rat_Circumvallate papillae

Get tips on using RIPA Buffer (10X) to perform Protein isolation Mammalian cells - Rat_Renal tissue

Protein isolation is a technique that involves isolation and/ or purification of protein from cells or tissues via chromatography or electrophoresis. The major challenges in protein isolation include: 1. The concentration of proteins in cells is variable and tends to be small for some intracellular proteins. Unlike nucleic acids, proteins cannot be amplified. 2. Proteins are more unstable than nucleic acids. They are easily denatured under suboptimal temperature, pH or salt concentrations. 3. Finally, no generalized technique/protocol can be applied for protein isolation. Proteins may have different electrostatic (number of positively or negatively charged amino acids) or hydrophobic properties. Therefore, protein purification requires multiple steps depending on their charge (a negatively charged resin/column for positively charged proteins and vice-versa), dissolution (using detergents) and unlike in the case of DNA and RNA, instead of using salts, proteins should be isolated by isoelectric precipitation.

Get tips on using RIPA Lysis and Extraction Buffer to perform Protein isolation Mammalian cells - Rat_Mesenteric fat