Glucose Assay Kit (ab65333)

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Overview

  • Product name
    Glucose Assay Kit
    See all Glucose kits
  • Detection method
    Colorimetric/Fluorometric
  • Sample type
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • Assay type
    Quantitative
  • Sensitivity
    1 µM
  • Range
    1 µM - 10000 µM
  • Assay time
    0h 40m
  • Product overview

    Glucose Assay Kit ab65333 is a rapid, simple and sensitive assay used to quantify glucose levels in biological samples such as serum, plasma, and other body fluids, food, growth medium, etc.


    In the glucose assay protocol, the glucose enzyme mix 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 amount of glucose.


    The kit detects glucose in the range 1-10000 µM.


    Glucose assay protocol summary:
    - add samples (deproteinized) and standards to wells
    - add reaction mix and incubate for 30 min at 37ºC
    - analyze with microplate reader

  • Notes

    If you have reducing substances in your samples, we recommend using Glucose Detection Kit II (ab102517).

    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Platform
    Microplate reader

Properties

Images

  • Functional studies- ab65333
    Functional studies- ab65333Image from Shao W et al., PLoS One 7(1), Fig 2C. Doi: 10.1371/journal.pone.0028784.

    Shao et al investigated the functional outcome of long- term curcumin supplementation on glucose homeostasis. Glucose metabolism was determined in animals with low fat diet (LFD), high fat diet (HFD) and HFD with curcumin feeding  using ab65333. Intraperitoneal insulin tolerance tests(IPITT) were conducted at the end of the 26 weeks. It was concluded curcumin improves insulin sensitivity and disposal of glucose. 

  • Functional Studies - Glucose Assay Kit (ab65333)
    Functional Studies - Glucose Assay Kit (ab65333)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - Glucose Assay Kit (ab65333)
    Functional Studies - Glucose Assay Kit (ab65333)

    Standard curve: mean of duplicates (+/- SD) with background reads subtracted

  • Functional Studies - Glucose Assay Kit (ab65333)
    Functional Studies - Glucose Assay Kit (ab65333)

    Glucose measured in cell lysates showing quantity (nmol) per million cells.

    Samples with the concentration of 2x107 cells/mL were used. Samples were diluted 1.5-13.5 fold and measured colorimetrically.

  • Functional Studies - Glucose Assay Kit (ab65333)
    Functional Studies - Glucose Assay Kit (ab65333)

    Glucose measured in human biological fluids showing quantity (µmol) per mL of tested sample. Samples were diluted 13.5 fold and measured colorimetrically.

Protocols

References

This product has been referenced in:
  • Stout AC  et al. Inflammation and glucose homeostasis are associated with specific structural features among adults without knee osteoarthritis: a cross-sectional study from the osteoarthritis initiative. BMC Musculoskelet Disord 19:1 (2018). Functional Studies . Read more (PubMed: 29304778) »
  • Williamson BK  et al. The Effects of Glaucoma Drainage Devices on Oxygen Tension, Glycolytic Metabolites, and Metabolomics Profile of Aqueous Humor in the Rabbit. Transl Vis Sci Technol 7:14 (2018). Read more (PubMed: 29423340) »
See all 31 Publications for this product

Publishing research using ab65333? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 32 Abreviews or Q&A

Question

I would like to confirm that this kit is not affected by the presence of phenol red in the media. I am using 1 ul of cell culture media with phenol red.

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Answer


The signal contributed by the phenol red in 1ul samples will be insignificant compared to the signal generated by the glucose in your samples. You should include a control consisting of 1ul of culture medium and subtract the absorbance or fluorometric value of that control from the values of your experimental samples.

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Question

Will peroxide interfere with this assay?

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Answer

Thank you for contacting us. Yes, peroxide is known to interfere with this Glucose Assay Kit. I hope this helps, please let me know if you need any additional information.

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Question

Can this kit distinguish between glucose and 2DG?

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Answer




This kit is only specific for D-glucose.

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Question

I want to do the glucose test on plasma from infected patients. As our plate reader is outside the BSL, we're not allowed to take out material without inactivating it. Could you tell me whether it is possible to inactivate samples with H2SO4 or PFA before using the test or after the color change has taken place or whether this will interfere with the assay or measurement?

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Answer

I have been in contact with the lab and unfortunately, we have not tested whether the addition of acids such as H2SO4 or PFA will interfere with the assay.

They have suggested that in this situation, perhaps it is best if you deproteinise your samples to remove most proteins/enzymes which would constitute the “biohazard” in your samples.

Alternatively, would 10% SDS be suitable for inactivating your samples? Although we have not tested this, itght work since it has been used in other assays with similar chemistry.

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Question

We need to measure free glucose in purified polysaccharide samples (bacterial derivation). Is kit ab65333 appropriate for use with our sample or will the enzyme break down larger polysaccharides, leading to false positive/high readings?

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Answer

We have not tested glucose in purified polysaccharide samples. The first step of the reaction is specific for glucose, so we expect this to work. It's possible that there will be some background, but this can be corrected for by subtracting the background control values (see page 8 of the protocol).

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Question

Our samples are not compatible with the oxidase/peroxidase-based glucose assay ab65333. What is the principle of the ab102517 assay?

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Answer

The main difference between these two glucose assays is that in ab65333 the probe reacts with hydrogen peroxide and gets oxidized to produce colored compound detectable at 570nm or at Ex/Em 535/587nm. In ab102517, the probe reacts with NADH (produced during glucose dehydrogenation in the first step) and forms a colored product measured at 450nm.

Assay ab65333 can measure 1uM-10mM glucose. The additional sensitivity in ab65333 is achieved in the fluorometric measurement of the assay. Assay ab102517 can measure concentrations within the range of 20uM-10mM.

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Question

I want to measure glucose being released into media. How should the glucose contained in media be removed prior to the measurements?

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Answer

I suggest exchanging the medium before collecting samples. That is, culture the cells in medium with a glucose/energy source and then exchange the medium with glucose-free medium such as Krebs-Ringer-Phosphate-HEPES (KRPH) buffer for example, and then run the glucose assay using the KRPH buffer. All the glucose in the buffer will be from the cells. There might be need for stimulation of glucose transporter for the cells to release glucose into the buffer. I would strongly suggest referring to publications on these types of experiments for protocols.

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Question

Two questions:
Question1:
Ab: 65333: Wants to confirm that the minimum sample volume is .5 uL that have a minimum of 1uM
Question 2:
Ab 65336: What is the minimum and maximum volume of serum that can be tested directly.

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Answer

Thank you for your enquiry.


Answer 1: Yes, the minimal serum volume for this assay is 0.5 µl. The lower detection limit of this assay is 1 µM. That does not mean that you have to start with a particular conc of the sample. More specifically, you would not know the conc of glucose to begin with.



Answer 2:

In the protocol step mammalian serum consists of 0.1-6 mM triglyceride. This is the conc of triglyceride, not serum. One can use a minimum of 1 µl and a max of 50 µl for this assay.

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Question

Inquiry: Dear Tech Support, I'm looking for the Abcam Glucose Assay Kit (ab65333). I've to use this kit to measure glucose consumption in various haematological cell lines and primary samples of AML and ALL. What are the concentrations that i should use in my experiments? 1x10^6 is sufficient or i need more cells?

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Answer

This kit would be used to measure glucose in the cell culture supernatant. The number of cells in culture would have an effect on the glucose consumption, but the amount of time the cells are cultured and the specific cell line you are testing for will also have an effect on glucose levels.

Therefore, it is difficult to provide precise details of the number of cells you should use. I can suggest growing the cells as normal, taking samples at various time points. In the protocol, we suggest testing several doses of your sample to make sure the readings are within the standard curve range. You may need to dilute them in order to ensure they are within range, in which case you will need to take this into account when calculating the glucose concentration from the reading obtained.

I am sorry it is difficult to be more precise than this for individual experiments, but I hope this will be helpful.

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Question

Respectable,

We have fermentation process and we need monitoring glucose in that process.
Expected concentracion of glucose is 0,1 - 3%.
We found two type test kits ( Glucose Assay Kit andGlucose Detection Kit) ). Please send me commercial offer for both and please recommendone that will match to our needs.
Best regards,

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Answer

Thank you for your enquiry which has been forwarded to the scientific support team.

I can confirm I would suggest Glucose Detection Kit ab102517or ab65333 Glucose Assay Kitswill be suitable for detection andquantification of glucose in the culture samples. I have put information below regarding the sensitivity and detection range of the kits, I can suggest to review this to decide which kit is best for your experiments (I am sorry we do not have concentration expressed as a percentage, I am not sure what this would be).

ab102517 has a senstivity of > 0.02 mM, range 0.02 mM - 10 mM
https://www.abcam.com/index.html?datasheet=102517 (or use the following: https://www.abcam.com/index.html?datasheet=102517).

ab65333 has a sensitivity of 1 µM, range 1 µM - 10000 µM
https://www.abcam.com/index.html?datasheet=65333 (or use the following: https://www.abcam.com/index.html?datasheet=65333).

I am sorry we do not sell directly to Russia. However, we do have an official distributor in Russia, Almabion. I can recommend to contact them directly to request a price quote:

ALMABION
ul. Vladimira Nevskogo d.59, k.166
Voronezh
394005
Russian Fed

Te l+7 4732 734828
Websitehttp://www.almabion.ru
Fax+7 4732 734828
mailto:Emailinfo@almabion.ru

I hope this will be helpful to you. Please do not hesitate to contact me if you have any further questions.

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