When it comes to mouse tissue samples, not all western blot loading control proteins are expressed equally. For example, there are relatively low amounts of β-actin in mouse heart and relatively low amounts of GAPDH in mouse small intestine. Plus, under certain experimental conditions, the protein levels of some loading control proteins can be affected. β-actin mRNA levels, for instance, can change with insulin treatments in some tissues.
Blog: How do I choose a loading control for my western blot?
Therefore, it is important to do research in advance and determine what the best loading control is for your sample type and experiment. Continue reading this blog post for example western blots showing the expression levels of common loading control proteins in various mouse tissues.
How does a Cell Signaling Technology (CST) scientist prepare for testing mouse tissues by western blot? We check reliable expression databases and the literature to ensure the tissue type expresses a loading control protein before doing a western blot. We also look to see what the relative abundance of that protein is to help set expectations. Just because a tissue expresses the mRNA for a given target does not mean the amount of resulting protein will be high enough to be detected by western blot. Make sure a given loading control protein is highly expressed in your tissue samples before running any blots so you can save valuable time and reagents.
Related Resource: Western Blot Troubleshooting Guide
In an effort to help, we have already performed testing in-house using some common loading control antibodies and mouse tissue types to serve as a reference for your experiments. The results are summarized in the reference table below.
| Brain | Spleen | Kidney | Pancreas | Testis | Skin | Eye | Heart | Lung | Skeletal Muscle | |
| α-Tubulin | ✔ | ✔ | ✔ | Low | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ |
| β-Actin | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | Low | ✔ | ✔ |
| β-Tubulin | ✔ | ✔ | ✔ | Low | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ |
| Cox IV | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ |
| GAPDH | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ | ✔ |
| Histone H3 | ✔ | ✔ | ✔ | ✔ | ✔ | Low | Low | ✔ | ✔ | ✔ |
| β3-Tubulin | ✔ | Low | Low | Low | ✔ | Low | ✔ | Low | ✔ | ✔ |
Table: Expression level of common loading control antibodies in various mouse tissue types.
Below, you'll also find example western blots showing the expression levels of common loading control proteins by mouse tissue type. You'll also find a list of the CST loading control antibodies that have been validated for use in western blot, and a brief list of antibodies recognizing each mouse target protein.
For the most up-to-date product availabilities and selection, please check out the complete CST product catalog. Our team is also happy to answer any purchasing questions.
WB analysis of extracts from various mouse tissues using α-Tubulin (11H10) Rabbit mAB #2125.
CST α-tubulin antibodies for western blot with mouse reactivity:
Note: β-Actin can be cleaved by caspase-3 between Asp244 and Gly245, which will result in a 32 kDa fragment and a 15 kDa fragment. Depending on the location of its antigen, some β-actin antibodies can detect both full-length β-actin as well as the 32 kDa fragment, also known as fraction.
WB analysis of extracts from various mouse tissues using β-Actin (D6A8) Rabbit mAB #8457.
CST β-actin antibodies for western blot with mouse reactivity:
WB analysis of extracts from various mouse tissues using β-tubulin (9F3) Rabbit mAb #2128.
CST β-tubulin antibodies for western blot with mouse reactivity:
WB analysis of extracts from various mouse tissues using COX IV Antibody #4844.
CST COX IV antibodies for western blot with mouse reactivity:
WB analysis of extracts from various mouse tissues using GAPDH (D16H11) XP Rabbit mAB #5174.
CST GAPDH antibodies for western blot with mouse reactivity:
WB analysis of extracts from various mouse tissues using Histone H3 (D1H2) XP Rabbit mAB #4499.
CST Histone H3 antibodies for western blot with mouse reactivity:
WB analysis of extracts from various mouse tissues using β3-Tubulin (D65A4) XP Rabbit mAB #5666.
CST β3-Tubulin antibodies for western blot with mouse reactivity:
Still not sure what the best antibody is for your experiment? Feel free to contact CST Technical Support for advice about choosing an antibody or setting up your experiment. We look forward to helping you choose the best antibodies for your experiments!
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