What is the secondary antibody for GAPDH?

Goat Anti-Rabbit IgG H&L
We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) secondary antibody. Use a concentration of 5 µg/ml. We recommend Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody.

Why is GAPDH used in Western blot?

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.

What is the kDa of GAPDH?

GAPDH molecule is a homotetramer composed of 36 kDa subunits. Thus the molecular weight for the whole molecule is 144 kDa.

What is the purpose of probing with GAPDH?

In Western blotting we often use GAPDH as a loading control. What this means is that by probing for GAPDH we can check that we have a loaded equivalent amounts of proteins on different lanes of the blot. An example of use – say we have a disease that we think causes an elevation of a particular protein in the cell.

What type of protein is GAPDH?

GAPDH is a homo tetramer containing four identical 37 kDa subunits. Localized in chromosome 12, human GAPDH gene transcribes single mRNA species, consequently producing a subunit that comprises a polypeptide chain of 335 amino acids7,8.

Why is GAPDH a housekeeping gene?

GAPDH is a key regulatory enzyme, which catalyzes the oxidative phosphorylation of glyceraldehyde-3-phosphate during glycolysis. β-actin is a cytoskeletal protein involved in cell structure and motility. Based on these basic and ubiquitous cellular functions, GAPDH and β-actin are considered as housekeeping genes.

Why is GAPDH used in PCR?

As one of the most common reference genes, GAPDH is often used to normalize the gene expression data, being used as an endogenous control in the quantitative analysis of RT-PCR, since in some experimental systems, its expression is very constant (Edwards and Denhardt 1985; Winer et al. 1999).

Is GAPDH a good loading control?

GAPDH can work as a reliable loading control only in Rhesus monkeys model of myocardial ischemia. Total protein level is the best loading control for western blot in animal models of myocardial ischemic infarction.

How many base pairs is GAPDH?

In order to understand the mechanism of this regulation, we have isolated and sequenced 5.4 kilobase pairs of a 12-kilobase pair human genomic clone encoding a functional GAPDH gene. The gene consists of 9 exons and 8 introns with eukaryotic signals necessary for the transcription and translation of GAPDH mRNA.

What type of control is GAPDH?

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), initially identified as a glycolytic enzyme and considered as a housekeeping gene, is widely used as an internal control in experiments on proteins, mRNA, and DNA.

What regulates GAPDH?

GAPDH is regulated by several cancer-related factors. Moreover, PTMs, such as phosphorylation, ADP ribosylation, and acetylation, are involved in processes by which cancer cells hijack normal pathways, as well as in cancer cell development, progression, and dissemination105.

Why is GAPDH a bad reference gene?

Studies have shown diverse functions and activity of this protein from DNA and RNA binding to key roles in neurodegenerative disease such as Alzheimer’s [3]. Therefore, the variable expression in these different cell types may result in GAPDH serving as a poor reference gene.