Beta-actin antibody – A control that matters
Actin is a major component of the cytoskeleton, with six known isoforms in mammals. Four of these isoforms are expressed primarily in striated and smooth muscle cells, whereas beta-actin and gamma-actin isoforms are ubiquitously expressed. Though beta-actin has a molecular weight of 42 kDa, degradation will produce 35–38 kDa, ~ 30 kDa, 15 kDa, and other typical fragments on a Western blot, of which 35-38 kDa fragments are the most common.
Proteintech’s monoclonal beta-actin antibody (60008-1-Ig) (Figure 1) recognizes all fragments, while 66009-1-Ig antibody (Figure 2) can only detect the full length of beta-actin. Beta-actin is highly conserved among species. However, the N-terminal sequence is less conserved among species than the C-terminus. Since the immunogen for 60008-1-Ig is the internal sequence, this antibody can recognize a wide range of species and can be broadly used in research.
|Figure 1||Figure 2|
|Figure 1. Immunofluorescent analysis of HepG2 cells using 60008-1-Ig (beta-actin Antibody) at a dilution of 1:50 and Rhodamine-labeled goat anti-mouse IgG (red).||Figure 2. Immunohistochemistry of paraffin-embedded human colon cancer tissue slide using 66009-1-Ig (beta-actin antibody) at a dilution of 1:1000 (under 40x lens). Heat-mediated antigen retrieved with Tris-EDTA buffer(pH9).|
Loading controls are essential for proper interpretation of Western blots. They can be used to normalize the levels of protein detected by confirming that protein loading is the same across the gel. In Western blotting, beta-actin (42 kDa) is commonly chosen as a loading control due to its broad and consistent expression across all eukaryotic cell types and the fact that expression levels of this protein are not affected by most experimental treatments (Figure 3).Beta-Actin runs as a “loading control” to show that very similar amounts of protein were loaded in each lane. This is essential in some experimental systems where it may not be possible to always collect the exact same number of cells for each sample, such as when working with primary cells, tissue samples, or systems where you may be altering protein synthesis, cell size/number, or apoptosis (1).
|Figure 3. Western blot analysis of beta-actin in various tissues and cell lines using Proteintech antibody 66009-1-Ig at a dilution of 1:20000 (Exposure time: 10 seconds).|
Beta-actin has been implicated in hand, foot, and mouth disease by proteomics and large diffuse B-cell lymphoma (3, 4).
|Antigen Name||Catalog No||
|alpha Actin||23660-1-AP||ELISA, WB, IHC, IF, IP||5|
|11032-1-AP||ELISA, WB, IP||1|
|23082-1-AP||ELISA, IHC, WB||1|
|alpha cardiac Actin-specific||66125-1-Ig||ELISA, WB, IHC||0|
|smooth muscle actin||14395-1-AP||ELISA, IHC, IP, WB, FC, IF||51|
|23081-1-AP||ELISA, WB, IHC, IP||5|
|55135-1-AP||ELISA, IHC, WB||4|
|gamma Actin||11227-1-AP||ELISA, WB||0|