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Facts about Aspartate aminotransferase, cytoplasmic.
The aspartate aminotransferase activity is involved in hepatic glucose synthesis during development and in adipocyte glyceroneogenesis. Using L-cysteine as substrate, regulates levels of mercaptopyruvate, an important source of hydrogen sulfide.
Human | |
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Gene Name: | GOT1 |
Uniprot: | P17174 |
Entrez: | 2805 |
Belongs to: |
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class-I pyridoxal-phosphate-dependent aminotransferase family |
aspartate aminotransferase, cytoplasmic; EC 2.6.1.1; GIG18; Glutamate oxaloacetate transaminase 1; glutamic-oxaloacetic transaminase 1, soluble (aspartate aminotransferase 1); growth-inhibiting protein 18; Transaminase A
Mass (kDA):
46.248 kDA
Human | |
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Location: | 10q24.2 |
Sequence: | 10; NC_000010.11 (99396870..99430624, complement) |
Cytoplasm.
Are you interested in Boster Bio's Anti-GOT1B GOLT1B Antibody? If so, you can read on to learn about its applications, ELISA kit kits, and high-affinity prima antibodies. You can also learn more about the product credit. These credits can be used by scientists all over the world. Here are a few:
GOT1 plays a critical role in cancer research. Antibodies against the GOT1 marker have a special affinity for human cancer cell lines and can detect different types of cancer cells. Boster Bio Anti GOT1B GOLT1B Antibody have been tested using Western blot, ELISA, and WB. It reacts with Human and Mouse cells.
The GOT1 gene encodes an enzyme that helps to export sialic acid free from lysosomes. Mutations to the gene can cause sialic-acid storage diseases, such as Salla and infantile SIACD (Itillic acid deposition disorder). GOT1 deficiency might be a target of therapeutics. This article focuses on several applications of GOT1 in therapeutics.
These experiments demonstrate that autophagy and iron levels are increased in orthotopic and subcutaneous cancers by knocking down GOT1. Also, GOT1 knockdown increases the sensitivity of the iron chelator DFO. FINO2 and ferric ammonium citrate potentiate the effect of GOT1 inhibition. GOT1 knockdown cells displaying autophagy are inhibited with calcein AM (FINO2) and calcein AM (calcein-AM).
GOT1 knockdown reduces PDA cell growth. This is a condition characterized as oxidative stress. GOT1 knockdown also enhances the effects of RSL3, a covalent inhibitor of GPX4, which induces ferroptosis. Inhibition of GOT1 increased intracellular cysteine and glutamate levels, while xCT protein was unaltered.
In vivo, GOT1 knockdown inhibits tumor proliferative ability. The PDA cells become firefly luciferase after 5 days of GOT1 knockdown. Inhibition of GOT1 also inhibits proliferation but does not cause cytotoxicity. Thus, GOT1 knockdown can be used for a variety of diagnostic purposes and drug development.
There are commercial ELISA products that can detect the GOT1 gene. They include reagents such as buffers and standards. A GOT1 ELISA set includes a sample diluent with detection antibody, capture antibody, streptavidin HRP, and a standard curve. It can also be used for testing different samples.
Reagents for competitive ELISA kit are the same as for direct ELISA. The reagents in an uncoated ELISA kit include the stop solution, wash buffer, and a diluted sample. The antibodies must be conjugated to a carrier protein. To avoid cross-reactivity between the antibodies and carrier proteins, they should not be stored at temperatures higher than 4°C or -20°C.
An ELISA kit for GOT1 identifies the enzyme responsible in the reaction between glycerol & glucose. The reagents can be used with samples from mice, rats, humans, and mice. GOT1 ELISA Kits can be used to detect cancer cells. GOT1 is an important regulator of nitrogen and carbon metabolism. Interestingly, its expression has been found to correlate with the growth of tumor cells.
ELISA technology has become a significant part of medical care and scientific research. It facilitates the development of new diagnostic tests and enables collaboration between different disciplines. ELISA technology has been a breakthrough in the diagnosis and treatment of HIV, Cancer, and Pregnancy. This marker is now more widely understood and the world is closer to finding a cure. With the advancement of diagnostic tests, we are more conscious of its role as saving lives.
It is essential to determine what type antigen will be used when selecting an ELISA test kit for the GOT1 mark. There are two types of sandwich ELISA kits. Sandwich ELISA uses two antibodies which recognize different epitopes. These are known as matched antibodies pairs. The capture antibody is coated on the multi-well plate, while the detection antibody is placed on the plate.
Biological marker are used to identify antigens with high affinity by recognizing them with high affinities. Primary antibodies are immunoglobulins, which bind to certain antigens. They can then be used for purification or detection. Primary antibodies are developed in animals as hosts. Some are monoclonal or others are polyclonal. In both cases, the antibodies detect a specific antibody.
In immunohistochemistry, high-affinity primary antibodies based on the GOT1 marker are widely used for molecular imaging. This antibody can recognize many antigens and detect posttranslational modifications. It can also identify proteins associated to various diseases. It is highly sensitive and long-lastingly active. It has an affinity constant that ranges between 10-6 and 10-9 for various antigens. Biotin, one of nature's strongest and most noncovalently bound bonds, is used to enhance the affinity for primary antibodies.
To obtain high-affinity primary antibodies, one must first prepare antigen-specific antibody preparations. This involves immobilizing the antigens on nitrocellulose and eluting them in acidic pH 2.0 glycine buffers. High-affinity antigens are not always desirable as it can be difficult to separate them. You must use optimal elution protocols.
The high-affinity primary antibodies can detect other antigen-specific proteins in addition to the GOT1 marker. These antibodies are vital for clean western-blot results. The antibodies detect proteins specific to the target antigen. However, antibodies may differ from one another, leading to inconsistent results. Additionally, a variety of epitopes may cause bands to be detected that are unspecific.
Secondary antibodies can be made using the GOT1 indicator by immobilizing serum proteins first and then lyophilizing them afterwards. Cross-adsorption is what is known and is highly recommended for applications that involve serum proteins and multiple primary antibody. This method is particularly helpful for detection of tumors and other samples in the bloodstream. Cross-adsorption comes with its own set of drawbacks. You can find out more about this process by reading the following.
This method involved transfecting 3T3-L1 cells with either an empty overexpression vector (or the GOT1 marker) with a 3T3-L1 cell line. After five days, protein lysates could be collected using RIPA buffer containing an inhibitor cocktail. Cells were then cultured another week. After this, differentiation was inducible and ORO staining took place 8 days later. Cells expressing GOT1 had higher levels of lipid accumulating. Furthermore, lipid accumulation was associated with an increase in the expression of PPARg and aP2.
To make secondary antibodies using the GOT1 marker, parental PDA cell lines were transduced with lentivirus containing shRNA plasmids at optimized viral titers. Stable cell lines were established after blasticidin and puromycin screening. Dmitri Wiederschain (Addgene) donated a TetpLKOpuro entry virus, which was used in the creation of dox-inducible GOT shRNAs.
After the preparation of the primary antibodies, secondary antibodies can then be conjugated to any number of labels. The downstream application will dictate the choice of label. Two commonly used enzymes to label secondary antibodies are horseradish peoxidase and alkaline-phosphatase. Alkaline Phosphatase is more stable and provides a longer-lasting signal. These secondary antibodies don't cost much.
PMID: 1974457 by Bousquet-Lemercier B., et al. Properties of human liver cytosolic aspartate aminotransferase mRNAs generated by alternative polyadenylation site selection.
PMID: 2241899 by Doyle J.M., et al. The amino acid sequence of cytosolic aspartate aminotransferase from human liver.
*More publications can be found for each product on its corresponding product page