製品の概要

  • 製品名

  • 検出方法

    Colorimetric/Fluorometric
  • サンプルの種類

    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids
  • アッセイタイプ

    Quantitative
  • 検出感度

    1 µM
  • 検出範囲

    1 µM - 10000 µM
  • 全工程の試験時間

    0h 40m
  • 製品の概要

    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

  • 特記事項

    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.

  • 試験プラットフォーム

    Microplate reader

製品の特性

画像

  • 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. 

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

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

  • 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.

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

プロトコール

参考文献

This product has been referenced in:

See all 45 Publications for this product

レビューと Q&A

1-10 of 32 Abreviews or Q&A

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.

Read More

Question
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.

Read More

Question
Answer




This kit is only specific for D-glucose.

Read More

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.

Read More

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).

Read More

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.

Read More

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.

Read More

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.

Read More

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.

Read More

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.

Read More

1-10 of 32 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

登録