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GST Fluorometric Activity Assay Kit

based on 2 citations in multiple journalsGST Fluorometric Activity Assay Kit24.1 4
Highly sensitive in vitro assay
Catalog #: K260

In stock

$345.00

Product Details

Cat # +Size K260-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 380/461 nm)
Species Reactivity Mammalian
Applications GST level in samples can be easily detected using a fluorometer or a 96-well fluorometric plate reader. The kit can detect GST activity in crude cell lysate or purified protein fraction, and also quantitate GST-tagged fusion protein. Detects < 0.5 mU.
Features & Benefits • Simple procedure; takes only ~1-2 hours
• Fast and convenient
Kit Components • GST Assay Buffer
• GST Sample Buffer
• MCB Substrate
• Glutathione (lyophilized)
• GST Standard (20 mU/µl)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Glutathione S-transferase (GST) is a family of enzymes that play an important role in detoxification of xenobiotics. GST catalyzes the formation of the thiol group of glutathione to electrophilic xenobiotics. It utilizes glutathione to scavenge potentially toxic compounds including those produced as a result of oxidative stress and is part of the defense mechanism against the mutagenic, carcinogenic and toxic effects of such compounds. The GST Fluorometric Activity Assay Kit provides a simple, fluorescence-based in vitro assay for detecting the GST activity using fluorescence plate reader. The assay utilizes monochlorobimane (MCB), a dye that reacts with glutathione. The free form of MCB is almost nonfluorescent, whereas the dye fluoresces blue (ex/em = 380/461 nm) when reacts with glutathione. GST catalyzes the MCB-glutathione reactions and the fluorescence levels are proportionally to the amounts of GST presence in the reaction. Thus, the GST level in samples can be easily detected using a fluorometer or a 96-well fluorometric plate reader. The kit can detect GST activity in crude cell lysate or purified protein fraction, and also quantitate GST-tagged fusion protein. Detects < 0.5 mU.


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We have a 405/8 excitation filter and a 535/25 emission filter - do you think that will work with the kit or do we need emission filter to be closer to 505?
Most plate readers have flexibility in their band width of detection. Your emission filter of 535/25 nm should work perfectly fine as long as it has the flexibility of +/- 20 nm.
How can I calculate the exact Cathepsin L levels in my samples with this kit?
This is a relative assay which will just show the fold increase of Cathepsin L between you induced and uninduced samples. To find the absolute levels of Cathepsin L in your sample, you will have to make a standard curve with free AFC (Cat # 1077).
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
 This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kits components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
How can we control auto-activation during the lysis and assay procedure?
The Cell lysis buffer will eventually lyse everything, however, only activated form can cleave the substrate. Autoactivation can be accounted for by using non-treated samples as a control
What is the sensitivity of this kit?
10-100 ng/assay
How do I use adherent cells with this protocol?
Wash your plate/wells with ice cold PBS once. Trypsinize the cells, and centrifuge them to get the cell pellet. Wash this pellet again with ice cold PBS and continue with the step IV.3.
Fletcher et al., Influence of glutathione-S-transferase (GST) inhibition on lung epithelial cell injury: role of oxidative stress and metabolism. Am J Physiol Lung Cell Mol Physiol, Jun 2015; 308: L1274 - L1285.
Marumo T et al (2008) J. Am. Soc. Nephrol. 10.1681/ASN.2007091040.
For more citations of this product click here
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