Glucose Colorimetric/Fluorometric Assay Kit

Sensitive Assay, HTS
Catalog #: K606 | abID: ab65333

Product Details

abID ab65333
Cat # +Size K606-100
Size 100 assays
Detection Method Absorbance (570 nm) or Fluorescence (Ex/Em 535/587 nm)
Species Reactivity Mammalian
Applications The kit detects 1-10000 µM glucose samples.
Features & Benefits • Simple procedure; takes ~40 minutes
• Fast and convenient
• Kit provides all necessary buffers and reagents for a accurate assay of glucose in various mammalian samples.
Kit Components • Glucose Assay Buffer
• Glucose Probe (in DMSO)
• Glucose Enzyme Mix (lyophilized)
• Glucose Standard (100 nmol/ml)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Glucose (C₆H₁₂O₆; FW: 180.16) is a very important fuel source to generate universal energy molecules ATP. Glucose level is a key diagnostic parameter for many metabolic disorders. Measurement of glucose can be very important in both research and drug discovery processes. The Glucose Assay Kit provides direct measurement of glucose in various biological samples (e.g., serum, plasma, body fluid, food, growth medium, etc.). Glucose Enzyme Mix specifically oxidizes glucose to generate a product which reacts with a dye to generate color (λ = 570 nm) and fluorescence (Ex/Em = 535/587 nm). The generated color and fluorescence is proportionally to the glucose amount. The method is rapid, simple, sensitive, and suitable for high throughput. The assay is also suitable for monitoring glucose level during fermentation and glucose feeding in protein expression processes. The kit detects 1-10000 µM glucose samples.

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Can I use this kit with tissue samples? If yes, can you provide a protocol for the sample preparation?
Yes, this kit can be used with tissue samples. Weigh 20-50 mg tissue, add 100-200 µl of cold assay buffer. Homogenize using a dounce homogenizer or by mild sonication on ice. Once the tissue is completely homogenized, centrifuge the homogenate at 10,000 g for 5 min. Take the supernatant and use it for the assay. We recommend deproteinizing the supernatant using a 10 kDa Spin column (BioVision Cat# 1997).
Will the phenol red in the media affect the assay readout?
Very low amounts of media are used for each sample. This will generate a very low background at the best. Please use only media as a background control and subtract this reading from all sample readings to accommodate for the phenol red.
Why does the final color development starts as pink, then goes to brown then disappears?
This is a very common phenomenon observed with use of the oxired probe and is caused due to excessive analyte concentration in the samples. Therefore, if your glucose samples are too concentrated, you may see this. Please dilute your samples with the assay buffer before reanalyzing.
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.
Can this kit be used with samples like bacteria, plants, drosophila 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 frozen tissue be used with this kit?
Although not ideal, flash-frozen tissue or cells stored at -80C can be used. It is important to remember that results may vary between fresh and frozen samples.
Is it possible to use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
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.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted
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.
Why are standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, the incubation time can be increased. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How are samples normalized against protein concentration?
A protein quantitation assay can be used on the supernatants from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can individual components of this kit be purchased?
Yes, any of the kit's components can be purchased without having to buy the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Does high sucrose concentration in the sample affect this assay's performance?
Yes, we have data comparing glucose in presence and absence of high sucrose (0.25M and 1M-effect almost identical) showing that the sucrose conc affects the assay. Therefore, this kit is not recommended to be used with samples.
What are the difference between the K606, K686 and K688 assay kit?
There are many differences between these 3 kits although they all measure the sample glucose concentration. A few of these differences which might make your decision of making a choice easier are:K606-100:Applicable for both colorimetric and fluorimetric detection.Requires filters for 570 nm and Ex/Em: 535/587 nm.Detetion limit of 1-10000 uM glucoseUtilizes the oxired probeK686-100Applicable only for colorimetric detection.Requires filters for 450 nm.Detetion limit of 20 uM to 10 mM glucoseDoes not utilize Oxired or Picoprobe (TM)Ideal for samples having reducing substancesK688-100Applicable only for fluorimetric detection.Requires filters for Ex/Em: 535/587 nm.Detetion limit of < 0.5 uM glucoseUtilizes the Picoprobe (TM)
Is it essential to make a standard curve for every expt, or is one curve/kit enough?
Yes, we strongly recommend you measure the standards every time you do an experiment. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
How can I prepare cell culture media for this assay?
When using cell culture supernatant (media), please spin down to get rid of the cell debris and use the supernatant for the assay. Cell culture media may not have as much proteins as seen in other samples types such as serum, plasma, etc. However, we still recommend to use the 10 kDa spin filter (Biovision Cat# 1997-25) to deproteinize the samples.
The highest detected concentration of this kit is at 10,000 μM. How is it calculated?
The highest limit of detection is calculated by dividing the highest Standard (10 nmol) by the lowest possible volume (1 µl in this case) = 10/1 = 10 nmol/µl = 10 mM = 10,000 µM.
Meredith A Sommars, Dynamic repression by BCL6 controls the genome-wide liver response to fasting and steatosis. eLife, April 2019;  30983568.
Reka Lorincz, In vivo monitoring of intracellular Ca2+ dynamics in the pancreatic β-cells of zebrafish embryos. Islets, Dec 2018; 30521410.
Ashley R. Maiuri, et al., (2018) Inflammation-induced DNA methylation of DNA polymerase gamma alters the metabolic profile of colon tumors, Cancer Metab, Jul. 2018, 30002826
Singh, Vikash et al. (2017) Salmonella Co-opts Host Cell Chaperone-mediated Autophagy for Intracellular Growth, J Biol Chem. 2017 Feb 3;292(5):1847-1864.
Yasuma, Taro et al. (2016) Amelioration of Diabetes by Protein S Diabetes. 2016 Jul;65(7):1940-51.
For more citations of this product click here