Anti-bodies

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 Ki67 antigen rabbit polyclonal antibody to perform Cell cytotoxicity / Proliferation assay cell type - malignant peripheral nerve sheath tumor

Get tips on using β-Actin Antibody #4967 to perform Western blotting β-Actin

Get tips on using alpha-Smooth Muscle Actin Antibody to perform Western blotting Smooth muscle actin

Get tips on using β-Actin Antibody (C4): sc-47778 to perform Western blotting β-Actin

Get tips on using NOTCH1/activated Notch1 Antibody to perform Immunohistochemistry Mouse - Notch1

Get tips on using Human/Mouse Active Caspase-3 Antibody to perform Western blotting Caspase-3

Get tips on using Smooth Muscle Actin Antibody (B4): sc-53142 to perform Immunohistochemistry Mouse - SMA

Get tips on using Active/Pro-Caspase 3 Monoclonal Antibody (31A1067) to perform Western blotting Caspase-3

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.