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Loading controls are usually proteins that exhibit high-level, constitutive expression in the cell type or sample you are studying. Housekeeping genes are often chosen for use this purpose. Loading controls are absolutely essential to ensure the reliability of your data when comparing expression of a protein in different samples.
Loading controls can be used to normalize the levels of protein detected by confirming that protein loading is the same across the gel. Here the main reasons you need to be using loading controls:
When lanes have not been loaded evenly, loading controls can be used to quantify the protein amounts in each lane by using the density of the loading control band to correct for the differences in loading.
Loading controls have a second role as a control in western blots. They can be used to check that there has been even transfer from the gel to the membrane across the whole gel. This is imperative when comparisons are made of the protein expression levels between samples.
This is an issue that is particularly important in signaling assays or experiments where a large number of lanes are being run at once. Proteins in the outer lanes of the gel are transferred to the membranes in a position close to the frame. This may result in more variation in binding compared to other areas of the gel.
Loading controls can show if this effect has occurred and allows the variation in binding to be corrected for.
Requested by referees
As an example, to be published in many Nature journals, a paper must meet four general criteria, the first of which is that it must "provide strong evidence for its conclusions." This directly correlates to the necessity of controls to prove that results obtained are valid.
Select a loading control that has a different molecular weight to the protein of interest. This ensures that you will be able to distinguish between the bands.
Use the table below to select the right loading control for your sample type.
Whatever your requirements, we have a loading control antibody to suit your experiment.
A wide range of targets
Select between 19 different targets for your research needs. Our loading control antibodies are perfect for multiple sample types, and cover molecular weights ranging from 15 kDa to 130 kDa.
Get sensitive and reproducible results with our loading control antibodies powered by RabMAb® technology.
We provide two pack sizes of 100 µL and 250 µL, and also offer a number of fully-validated HRP-conjugated loading controls.
Need more advice? Our experienced Scientific Support team is on hand to help.