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Product Details
| Cat # +Size | K321-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Absorbance (OD 293 nm) |
| Applications | Detection of Adenosine Deaminase activity |
| Features & Benefits | • Rapid, and convenient • The kit measures total Activity of Adenosine Deaminase with limit of quantification 1 mU recombinant Adenosine Deaminase. |
| Kit Components | • ADA Assay Buffer (10x) • ADA Convertor • ADA Developer • ADA Substrate • ADA Positive Control • Inosine Standard (10 mM) • U.V. transparent plate (96-well) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
Adenosine Deaminase (ADA) (E.C. 3.5.4.4.) is an enzyme that catalyzes the conversion of adenosine and 2’-deoxyadenosine to inosine and 2’-deoxyinosine. Adenosine Deaminase is widely distributed in various tissues and cells. There are two isoforms, ADA1 and ADA2. ADA1 is widely expressed in most cells in the body, particularly in lymphocytes and macrophages. It is present in the cytosol, nucleus and found associated with dipeptidyl peptidase-4 on the cell membrane. ADA2 was first found in the spleen but is predominantly found in the plasma and serum. Increased serum ADA levels are found in certain infectious diseases such as tuberculosis and various liver diseases such as acute hepatitis, alcoholic hepatic fibrosis, chronic active hepatitis to name a few. Adenosine Deaminase is also a marker for T-lymphocyte proliferation. In BioVision’s ADA Activity Assay, inosine formed from the breakdown of adenosine is converted to uric acid with ADA Convertor and ADA Developer. The uric acid is measured at OD 293 nm. The kit measures total Activity of Adenosine Deaminase with limit of quantification of 1 mU recombinant Adenosine Deaminase.
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Is the acetylated substrate from the cells?
The substrate is provided as a kit component. For easy understanding of the reaction, we have mentioned that it is an acetylated substrate bound to AMC.
What would be a “proper solvent” for the inhibitor? Will any liquid work? Also, what if the inhibitor is already in solution? I might want to test a cell or tissue extract
The type of solvent would depend on the inhibitor you use. For eg., Sirtinol (Cat # 2062-1,-5) can be dissolved in DMSO, ethanol and dimethyl formamide. The assay buffer in which you dilute it will act as the negative control. You can run the solvent alone as another negative control. Your inhibitor in the solvent will act as your expt samples. If you want to use inhibitors added to cell extracts, the cell extract itself will go in the place of the solvent as the solvent control.
Does the Inhibitor Control (Nicotinamide) already contain Assay Buffer? If not, it should be diluted with the same amount of Assay Buffer as the test samples
You can use the nicotinamide control as is. It does not need to be further diluted.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
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. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
