Western Blotting is an analytical technique used to detect specific proteins in a given sample. It uses gel electrophoresis to separate native or denatured proteins by the length of the polypeptide. The proteins are then transferred to a membrane where they are detected using antibodies specific to the target protein.
Real-time PCR, also called quantitative real time PCR (Q-PCR/qPCR), is used to amplify and simultaneously quantify a targeted DNA molecule. It enables both detection and quantification (as absolute number of copies or relative amount when normalized to DNA input or additional normalizing genes) of one or more specific sequences in a DNA sample.
Reverse transcription PCR includes two steps. The first step is reverse transcription, in which RNA is reverse transcribed to its DNA complement (complementary DNA, or cDNA) using reverse transcriptase and primers. The second step is amplification using traditional or real-time PCR.
Immunohistochemistry is a method of detecting the presence of specific proteins in cells or tissues. It is widely used in basic research to understand the distribution and localization of biomarkers and differentially expressed proteins in different parts of a biological tissue.
The indirect ELISA is used primarily to determine the strength and/or amount of antibody response in a sample. In the assay, the antigen of interested is immobilized by direct adsorption to the assay plate. Detection of the antigen can then be performed by using a matched set of primary antibody and conjugated secondary antibodies.
Polymerase Chain Reaction (PCR) is a technique to amplify few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. There are three major steps (Denaturation, Annealing and Extension) in a PCR and repeat for 30 or 40 cycles.
H&E stain is a popular staining method in histology. Its a combination of two dyes: the basic dye (hematoxylin) and the alcohol-based dye (eosin). In an H&E stain you will usually see both eosinophilia and basophilia: the nuclei of cells basophilic (blue), while eosinophilia is typical of cytoplasmic constituents (pink).
Cell culture is the process by which cells are grown under controlled conditions. This video shows you how to culture attached cells from cell thawing, cell maintenance, to cell freezing.
Immunofluorescence is a technique to visualize a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye. We use the indirect immunofluorescence staining to perform cells fixed on slides and examine under a fluorescence microscope.
In competitive ELISA, unlabeled antibody is incubated in the presence of its antigen. Then these bound antibody/antigen complexes are then added to an antigen coated well. After washing, unbound antibodies are removed. The more analytes in the sample, the less antibodies will be able to bind to antigens in the well. The signal is then detected using labeled secondary antibodies and the decrease in signal is compared to a control. The major advantage of a competitive ELISA is the ability to use crude or impure samples and still selectively bind any antigen that may be present.