GAPDH antibody, is usually used as a loading control antibody for Western Blot to normalize the levels of protein detected by confirming that protein loading is the same across the gel.
Contents
- 1 What is the purpose of GAPDH?
- 2 Why GAPDH is used as control in Western blot?
- 3 Is GAPDH a good loading control?
- 4 Why is GAPDH a housekeeping gene?
- 5 What are the other physiological functions of GAPDH?
- 6 Is GAPDH an antibody?
- 7 Why is B actin used as control?
- 8 Where does GAPDH run?
- 9 Where is GAPDH?
- 10 Is GAPDH a good housekeeping gene?
- 11 How big is GAPDH?
- 12 What is the purpose of a housekeeping gene?
- 13 What is protein loading control?
- 14 What is endogenous control?
What is the purpose of GAPDH?
Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is an enzyme involved in breaking down glucose to obtain energy. More specifically, in eukaryotes, this enzyme catalyzes the sixth step in glycolysis, converting glyceraldehyde 3-phosphate to D-glycerate 1,3-bisphosphate (1,3-BPG).
Why GAPDH is used as control in Western blot?
It is one of the so called housekeeping proteins and GAPDH is constitutively expressed in almost all tissues in high amounts. For this reason, GAPDH is widely used as a loading control for protein normalization in Western blotting.
Is GAPDH a good loading control?
GAPDH (36 kDa) is integral for glycolysis and plays many roles in nuclear function; such as transcription regulation and apoptosis. The stable and ubiquitous expression of GAPDH also make it a suitable loading control for many experiments.
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.
What are the other physiological functions of GAPDH?
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is an energy metabolism-related enzyme in the glycolytic pathway. Recently, it has been reported that GAPDH has other physiological functions, such as apoptosis, DNA repair and autophagy.
Is GAPDH an antibody?
This GAPDH antibody can be used as a loading control antibody. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. Preliminary data indicates that the GAPDH antibody- loading control ab8245 recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.
Why is B actin used as control?
Beta-actin, also known as a “housekeeping” protein, is usually used as a loading control, for among others, the integrity of cells, protein degradation, in PCR and Western blotting. They can be used to normalize the levels of protein detected by confirming that protein loading is the same across the gel.
Where does GAPDH run?
Transcription and apoptosis GAPDH moves between the cytosol and the nucleus and may thus link the metabolic state to gene transcription.
Where is GAPDH?
In addition to the cytoplasm where majority of GAPDH is located under the basal condition, GAPDH is also found in the particulate fractions, such as the nucleus, the mitochondria, and the small vesicular fractions. When cells are exposed to various stressors, dynamic subcellular re-distribution of GAPDH occurs.
Is GAPDH a good housekeeping gene?
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 western blots.
How big is 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 a housekeeping gene?
In molecular biology, housekeeping genes are typically constitutive genes that are required for the maintenance of basic cellular function, and are expressed in all cells of an organism under normal and patho-physiological conditions.
What is protein loading control?
Loading Control: A loading control is an antibody specific for a ubiquitously and constitutively expressed protein and can be used to normalize protein levels in Western blot. Loading controls can be general whole cell proteins or specific markers for subcellular fractions (e.g. nucleus, mitochondria, membrane).
What is endogenous control?
An endogenous control gene is a gene whose expression level should not differ between samples, such as a housekeeping or maintenance gene.
