|Product name||Human L-Glucosamine--fructose-6-phosphate aminotransferase (GFAT) ELISA Kit|
|Applications notes||This Human L-Glucosamine--fructose-6-phosphate aminotransferase (GFAT) ELISA Kit employs a two-site sandwich ELISA to quantitate GFAT in samples. An antibody specific for GFAT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyGFAT present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for GFAT is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of GFAT bound in the initial step. The color development is stopped and the intensity of the color is measured.|
|SampleType||Cell culture supernatants, Other biological fluids, Plasma, Serum|
|Assay type||Sandwich ELISA (quantitative)|
|Assay duration||Multiple steps standard sandwich ELISA assay with a working time of 3-5 hours. It depends on the experience of the operation person.|
|Kit components||• Human L-Glucosamine--fructose-6-phosphate aminotransferase microplate
• Human L-Glucosamine--fructose-6-phosphate aminotransferase standard
• Human L-Glucosamine--fructose-6-phosphate aminotransferase detect antibody
• Standard diluent
• Assay buffer
• HRP substrate
• Stop solution
• Wash buffer
• Plate covers
|Features & Benefits||Human L-Glucosamine--fructose-6-phosphate aminotransferase (GFAT) ELISA Kit has high sensitivity and excellent specificity for detection of Human GFAT. No significant cross-reactivity or interference between Human GFAT and analogues was observed.|
|Calibration range||Please inquire|
|Limit of detection||Please inquire|
|Usage notes||• Do not mix components from different kit lots or use reagents beyond the kit expiration date.
• Allow all reagents to warm to room temperature for at least 30 minutes before opening.
• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.
• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.
• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.
• It is recommended that all samples and standards be assayed in duplicate or triplicate.
|Storage instructions||The unopened kit should be stored at 2 - 8°C. After opening, please store refer to protocols.|
|Shipping||Gel pack with blue ice.|
|Precautions||The product listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.|
|Background||Glutamine:fructose-6-phosphate amidotransferase (GFAT) catalyzes the formation of glucosamine 6-phosphate and is the first and rate-limiting enzyme of the hexosamine biosynthetic pathway. GFAT controls the flux of glucose into the hexosamine pathway, and thus formation of hexosamine products, and is most likely involved in regulating the availability of precursors for N- and O-linked glycosylation of proteins. Expression of these cDNA in E. coli produced a protein of approximately 77 kD and increased GFAT activity 4.5-fold over endogenous bacterial levels. Recombinant.GFAT1L protein possessed functional GFAT activities and biochemical characteristics similar to those of GFAT1. Previously, GFAT1 was considered a simplex enzyme.|
Fig.1. Human L-Glucosamine--fructose-6-phosphate aminotransferase (GFAT) Standard Curve.
Fig.2. Abbkine ELISA kit is series of sandwich ELISA to quantitate specific protein in samples.