GAPDH Mouse Monoclonal Antibody #abs137959

abs137959-50ug

In Stock

201

abs137959-100ug

In Stock

301

Description

As a critical enzyme in the glycolytic pathway, glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is widely expressed in most cell types. Consequently, GAPDH antibodies are commonly employed as loading controls for Western Blotting. However, in certain conditions, such as hypoxia and diabetes, the expression levels of GAPDH may be affected and fluctuate in certain cell types. Despite this, the crucial role of GAPDH as an enzyme in cellular metabolism remains unchanged.

Source

Mouse

Specificity

The monoclonal antibody against GAPDH in mice is capable of detecting the protein at its endogenous levels. This antibody enables the detection of total GAPDH protein in samples. By binding specifically to this protein, the mouse monoclonal antibody allows for precise and accurate measurements of GAPDH levels, making it a valuable tool in research.

Species Reactivity

We have a diverse range of specimens in our study, including mammals such as humans, mice, rats, pigs, dogs, monkeys, hamsters, and even chickens. We also have some aquatic organisms like zebrafish, as well as farm animals like cows, sheep, goats, and rabbits. Additionally, we're studying plants such as rice and fish to explore their genetic and cellular makeup. With this broad range of subjects, we're hopeful to uncover valuable insights that can benefit numerous fields.

Application

For Western blotting, it is recommended to use a dilution ratio of 1:3000 to 1:10000. When performing immunohistochemistry, a dilution ratio of 1:200 is optimal. For immunofluorescence and immunocytochemistry experiments, a dilution ratio of 1:200 is also recommended. For ELISA assays using peptide antigens, the recommended dilution range is between 1:20000 and 1:40000. It is important to carefully consider the appropriate dilution ratio for each application to achieve accurate and reliable results.

Immunogen

Full-length GAPDH protein of human.

Properties

Concentration

1mg/ml

Purification

Affinity-chromatography.

Clonality

Monoclonal Antibody

Stability & Storage

To ensure the preservation of the product, it is recommended to store it at a temperature of -20 °C for a duration of one year. It is important to avoid subjecting the product to repeated freeze/thaw cycles.

Storage buffer

This is a description of Mouse IgG1, which is stored in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, with 150mM NaCl, 0.02% sodium azide, and 50% glycerol at a temperature of -20 °C. The product is stable for a duration of 12 months starting from the date of receipt.

Target

Background

This multifunctional protein has two distinct activities: glyceraldehyde-3-phosphate dehydrogenase and nitrosylase. Its participation in glycolysis is due to the former activity, while its nuclear functions are attributed to its nitrosylase function. In the nucleus, this protein is involved in various events such as transcription, RNA transport, DNA replication, and apoptosis.
The nitrosylase activity of this protein is responsible for the cysteine S-nitrosylation of nuclear proteins SIRT1, HDAC2, and PRKDC, which modulates their activity. This protein also plays a crucial role in the organization and assembly of the cytoskeleton by facilitating microtubule and membrane associations. Its ability to stimulate the binding of CHP1 to microtubules is essential for the CHP1-dependent microtubule and membrane interactions.
Additionally, this protein is a key enzyme in glycolysis that catalyzes the conversion of D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. It also participates in inflammation processes by forming the GAIT (gamma interferon-activated inhibitor of translation) complex, which mediates interferon-gamma-induced transcript-selective translation inhibition. The GAIT complex binds to stem-loop-containing GAIT elements in the 3' -UTR of various inflammatory mRNAs, such as ceruplasmin, suppressing their translation.
Overall, this multifunctional protein plays integral roles in both metabolic and regulatory processes, making it a crucial component of cellular function.

Posttranslational modification

S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus (By similarity). S-nitrosylation of Cys-247 is induced by interferon-gamma and LDL(ox) implicating the iNOS-S100A8/9 transnitrosylase complex and seems to prevent interaction with phosphorylated RPL13A and to interfere with GAIT complex activity.ISGylated.Sulfhydration at Cys-152 increases catalytic activity.Oxidative stress can promote the formation of high molecular weight disulfide-linked GAPDH aggregates, through a process called nucleocytoplasmic coagulation. Such aggregates can be observed in vivo in the affected tissues of patients with Alzheimer disease or alcoholic liver cirrhosis, or in cell cultures during necrosis. Oxidation at Met-46 may play a pivotal role in the formation of these insoluble structures. This modification has been detected in vitro following treatment with free radical donor (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide. It has been proposed to destabilize nearby residues, increasing the likelihood of secondary oxidative damages, including oxidation of Tyr-45 and Met-105. This cascade of oxidations may augment GAPDH misfolding, leading to intermolecular disulfide cross-linking and aggregation.

Celluar localization

Cytoskeleton;Cytosol;Extracellular region or secreted;Nucleus;Plasma Membrane;

UniPort

P04406