The use of Beta Actin (AC-15) as a loading control across multiple species

Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two isoforms that are highly and ubiquitously expressed in the cell.  For this reason, measuring beta-actin levels has served as a useful control in research experiments in order to have a baseline of protein expression to compare cell manipulations to.  However, beta actin has other implications in scientific research aside from acting

Tips on choosing an ideal loading control antibody for Western Blotting

Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification.

GAPDH - A "housekeeping" gene with diverse functions in cellular homeostasis

Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a well-known housekeeping gene with functions in glycolysis. Many biologists are familiar with the gene and use GAPDH antibodies for a loading control when performing west blots. However, this primarily cytoplasmic protein is an essential metabolic regulator and has been shown to be involved in a variety of cellular processes like DNA repair, membrane fusion, and cell death (1). Cytoplasmic GAPDH exists as a tetramer and normally mediates the formation of ATP and NADH during glycolysis (1).

A New Standard in Antibody Testing - Simple Western Certified Antibodies

The Western blot is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. The traditional Western blot can be a labor-intensive and time-consuming process, leading many researchers to seek an alternative method that is more efficient, reproducible and quantitative.

GAPDH: More than a housekeeping gene

GAPDH is a 146kD tetramer glycolytic pathway metabolic enzyme composed of four 30-40 kDa subunits. It is responsible for reversibly phosphorylating its substrate glyceraldehyde 3-phosphate within the glycolytic pathway.  Apart from its role in glycolysis, GAPDH may have other roles such as transcriptional activation. Due to its housekeeping role, GAPDH is highly expressed in almost all tissues, allowing its use as an internal loading control (traditionally for mRNA expression comparisons, but also in protein studies.

GAPDH (Glyceraldehyde 3-Phosphate Dehydrogenase)

GAPDH is a 146 kD tetramer glycolytic pathway metabolic enzyme responsible for reversibly phosphorylating glyceraldehyde 3-phosphate. It may have other possible functions in transcriptional activation. GAPDH is highly expressed due to this housekeeping role, and its prevalent expression has allowed its use as an internal loading control – traditionally for mRNA expression comparisons – but also in protein studies.

Time to Start Actin Like a Reliable 'Housekeeper'!

A growing body of data and studies using actin antibodies supports a view of the actin cytoskeleton of smooth muscle cells as a dynamic structure that plays an integral role in regulating the development of mechanical tension and the material properties of smooth muscle tissues.

Beta Actin and GAPDH: The Importance of Loading Controls

Western blotting is an essential technique to probe protein expression in complex cell or tissue lysates. To accurately determine protein expression and interpret Western blot results, it is important to use loading controls. 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.

The GAPDH Antibody in Western blot Assays

The loading controls on our antibody database are widely used in gel electrophoresis and Western blotting studies. Products like the GAPDH antibody detect "housekeeping" proteins which are abundantly distributed in cells. This makes them useful for checking the even loading of gel samples, and the even transfer of proteins at the blotting stage. They also serve a purpose in quality control, by verifying reagents are working correctly, and in the standardization of experimental results.

New Primers Available for ChIP Procedures

Novus has recently released six new primer sets designed for use in real-time PCR DNA amplification of housekeeping, silent, or heterochromatin associated proteins.