Dear friends, do you feel at a loss when facing various Loading Control and Epitope Tagging Antibodies during protein immunoblotting? Loading Control proteins are encoded by housekeeping genes in organisms and are stably expressed in multiple tissues. They are used to verify whether immunological experiments are proceeding normally and as a semi-quantitative detection standard for protein expression. So, how do you correctly choose loading control antibodies? You can refer to the following points.

I. Sample Species Origin

1. For samples from mammalian tissues or cells, generally choose β-actin, α-tubulin, GAPDH, Lamin-B, Histone H3, Na+/K+-ATPase.
2. For plant-derived samples, generally choose plant actin, Rubisco, etc.
3. For other samples with less research, choose the appropriate protein as an internal reference by consulting relevant literature or databases.

Ensure that the molecular weight of the target protein differs from that of the loading control by more than 5KDa to prevent interference during detection. For example, if the target protein is 40KDa, it is not suitable to choose β-actin (42KDa) as an loading control. Consider using GAPDH (36KDa) or β-tubulin (55KDa) as loading control.

III. Expression Site of the Internal Reference Protein

Label Antibodies Work Better When Used Together

In experiments studying protein interactions, such as pull-down, epitope tags can be fused to the N or C terminus of the target protein through molecular biological methods. Commonly used epitope tags include His, HA, Myc, and GST. These epitope tags usually do not affect the biological activity and intracellular localization of the target protein. The emergence of epitope tags has provided convenience for scientific research and industrial production.

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