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GAPDH Activity Assay Kit

based on 1 citations in multiple journalsGAPDH Activity Assay Kit14.1 4
Sensitive Assay, HTS
Catalog #: K680

In stock

$175.00

Product Details

Cat # +Size K680-100
Size 100 assays
Detection Method Absorbance (450 nm)
Applications Measurement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity in various tissues/cells
Analysis of glycolysis and pentose phosphate pathways
Features & Benefits • Simple, rapid & convenient
• can measure GAPDH activityas low as 100 µU/mL in a variety of samples
Kit Components • GAPDH Assay Buffer
• GAPDH Substrate
• GAPDH Developer
• NADH Standard (500 nmol)
• GAPDH Positive Control
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase; EC 1.2.1.12) catalyzes the conversion of Glyceraldehyde-3-Phosphate (GAP) to 1, 3-Bisphosphate Glycerate (BPG) and plays a key role in glycolysis. The enzyme is involved in cellular processes such as apoptosis, membrane trafficking, iron metabolism and nuclear translocation. GAPDH (housekeeping gene) expression is stable and constitutive. Deregulation of GAPDH activity is associated with abnormal cell proliferation and carcinogenesis. Accurate quantitation of GAPDH activity is important for diagnosing diseases and studying normal cellular physiology. BioVision’s GAPDH Activity Assay Kit provides a simple and sensitive method for monitoring GAPDH activity in various samples. In this assay, GAPDH catalyzes conversion of GAP into BPG and an intermediate, which reacts with a developer to form a colored product that absorbs maximally at 450 nm. Our high-throughput adaptable assay can detect GAPDH activity as low as 100 µU/mL in a variety of samples.


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How to prepare tissue lysates for the assay?
Here’s a general protocol to preparing tissue lysates: Start with 10-20 mg of the muscle tissue, add 200-500 µl (or ~4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Use the supernatant for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve.
DO you use Baculovirus expression system to prepare any component in the kit?
None of the components are from Baculovirus.
Can we use frozen samples with this assay?
We would recommend using fresh samples. However, frozen samples can be used too provided they have not undergone multiple freeze thaw cycles.
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only.
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?
BioVisions’s assay kits expire 6-12 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet.
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.
Reisz, Julie A. et al. (2017) Oxidative modifications of glyceraldehyde 3-phosphate dehydrogenase regulate metabolic reprogramming of stored red blood cells, Blood. 2016 Sep 22;128(12):e32-42.
For more citations of this product click here