Loading Controls Antibodies
In Western Blot experiments, inaccuracies in total protein concentration measurement or errors in protein sample loading due to operational issues may be present; these errors can be corrected by determining the actual amount of internal control protein transferred to the membrane in each sample.
Internal reference involves selecting a protein expressed from an endogenous source within the same sample as a reference. Housekeeping proteins are commonly chosen as internal controls, as they exhibit relatively constant expression across various tissues and cells, serving as a stable reference point for detecting changes in protein expression levels (such as GAPDH, β-Actin, etc.).
In WB experiments, protein expression is reflected through binding with antibodies, meaning that the choice of internal control protein is equivalent to the choice of internal control antibody.
Tag antibody
In biotechnology, DNA recombination techniques are often used to construct fusion proteins containing the target gene and epitope tags, known as tagged fusion proteins, which are then identified and purified using antibodies that can specifically recognize the tag. The principle of the entire reaction is based on antigen-antibody interactions, where tag antibodies can bind highly specifically to the corresponding tagged fusion proteins.
Fusion tags can simplify the purification process of proteins, control the spatial orientation of the fixed proteins, facilitate detection, visualize biological events in vivo, enhance the yield of recombinant proteins, and improve the solubility and stability of recombinant proteins, among other benefits. Commonly used tags include Flag, His, myc, HA, GST, GFP, etc.
Tag antibodies can be used to detect or purify the tag sequences on expression vectors, thereby studying the expression levels and functions of target proteins. Tag antibodies have now become common tools in research related to gene expression and gene function signal transduction.
