siRNA / RNAi /miRNA transfection Human Cells THP-1

Get tips on using RNAprotect Cell Reagent (250 ml) to perform RNA stabilization Cell lines

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - rat nucleus pulposus

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse cardiac fibroblasts

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - mouse embryonic fibroblasts

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - MIA PaCa-2

Get tips on using Senescence β-Galactosidase Staining Kit - Cell Signaling to perform Reporter gene assay β-galactosidase substrates - MDA-MB-231

Get tips on using Gibco™ RPMI 1640 Medium to perform 3D Cell Culture Media Mouse thymic organoids

Western blotting is a widely used technique to size separate proteins from a pool of cell or tissue lysates. The technique has 4 major steps: a) gel electrophoresis, b) blocking and treatment with antigen specific antibody, c) treatment with secondary antibody and finally d) detection and visualization. Though western blotting is a widely used technique, detection of specific proteins depends on several factors, the major ones are antibody concentration, incubation time and washing steps. Key points for obtaining clean blots are: always prepare fresh buffer solutions and optimize antibody concentration. Given the advent of high-throughput protein analysis and a push to limit the use of lab consumables, onestep antibodies are developed which recognise protein of interest and also contain a detection label.

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.