Beta actin western blot plus#

The expression levels of this protein do not vary drastically due to cellular treatment, which is another reason the protein makes a suitable control. Western blot analysis using anti-Beta Actin mAb against NIH/3T3 (1), Jurkat (2), Hela (3), CHO (4), PC12 (5), HEK293 (6), COS (7), A549 (8) and MCF-7 (9) cell lysates.īeta-Actin (42 kDa) is commonly chosen as a loading control due to its general expression across all eukaryotic cell types. The most popular loading control targets include Beta-Actin and GAPDH. Additionally, the loading control protein should demonstrate ubiquitous expression. When considering a loading control for your experiment, it is important to choose a protein that does not have a similar molecular weight to your target of interest. A loading control antibody helps determine if samples have been loaded equally across all wells and confirms effective protein transfer during the western blot protocol. To accurately determine protein expression and interpret Western blot results, it is important to use loading controls. Sufficient quantity of each antibody is included for several mini-blots.Western blotting is an essential technique to probe protein expression in complex cell or tissue lysates.

Beta actin western blot plus#

Set of four primary antibodies (beta-Actin, beta-Tubulin, GAPDH, Histone H3) specific to proteins frequently used for protein normalization, plus one HRP conjugated secondary antibody. Histone H2B nuclear loading control antibodiesĬaution: Amounts of histone doubles prior to cell division! Not suitable for comparing S-phase with with pre-S-phase cells. The VDAC/Porin antibodies are good alternative mitochondrial loading controls for proteins of this size. COX-4ĬOX-4 mitochondrial loading control antibodiesĬOX-4 is expressed at consistently high levels and thus makes a good loading control.Ĭaution: Many proteins run at the same 17 kD size as COX-4. VDAC is expressed at steady-state levels and thus makes a good loading control. VDAC/Porin mitochondrial loading control antibodies TubulinĬaution: Tubulin expression is known to vary according to resistance to antimicrobial and antimitotic drugs (Sangrajrang S. GAPDH is constitutively expressed in almost all tissues at high levels, therefore becoming the marker of choice as loading control in northern/western blots and protein normalization.Ĭaution: Some physiological factors, such as hypoxia and diabetes as well as some sort of cancers increase GAPDH expression in certain cell types. Suitability: Cytoplasmic and whole-cell extracts Suitability: Cytoplasmic and whole cell extractsĬaution: Changes in cell-growth conditions and interactions with extracellular matrix components have been found to alter actin protein synthesis (Farmer et al., 1983 Ben Zeev and Amsterdam,1986). It is also important that the protein chosen as a loading control has a different molecular weight than the protein of interest so that the bands are distinct and expression levels quantifiable. Thus “housekeeping genes” are frequently chosen for use as loading controls. Loading controls are usually proteins that exhibit high-level, constitutive expression in the cell type or sample you are examining. The expression levels of the loading control should not vary between the different sample lanes. They are used to normalize the levels of protein detected by confirming that protein loading is the same across the gel.

Loading controls are essential for proper interpretation of western blots.