siRNA / miRNA gene silencing Human U937 MK2 (MAPK Kinase 2)

Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - THP-1 Lipofectamine

Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - NK-92 Lipofectamine

Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - KG-1 Lipofectamine

Get tips on using Lipofectamine® RNAiMAX Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - Jurkat cells Lipofectamine

Get tips on using TransIT®-LT1 Transfection Reagent to perform siRNA / RNAi /miRNA transfection Human Cells - Cal 27 cells Polymer / lipid

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.

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.

Get tips on using MAP LC3β Antibody (G-2) to perform Autophagy assay cell type - AR42J

Get tips on using 2-D Quant Kit to perform Protein quantification Colorimetric method

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.