Immunohistochemistry Estrogen receptor (ER)

Get tips on using Estrogen Receptor α (D8H8) Rabbit mAb #8644 to perform ChIP Anti-bodies ERα

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.

Get tips on using Recombinant Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) to perform Western blotting Estrogen Receptor

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 Recombinant Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) to perform ChIP Anti-bodies ERα

Get tips on using TransAM® ER Transcription Factor ELISA Kits to perform ELISA Human - Estrogen receptor (ESRs)

Get tips on using Human Total ER alpha/NR3A1 DuoSet IC ELISA to perform ELISA Human - Estrogen receptor (ESRs)

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.