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    free-glycerol-assay-kit-ab65337.pdf

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Free Glycerol Assay Kit (ab65337)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (10)References (16)

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Functional Studies - Free Glycerol Assay Kit (ab65337)
  • Functional Studies - Free Glycerol Assay Kit (ab65337)
  • Functional Studies - Free Glycerol Assay Kit (ab65337)
  • Functional Studies - Free Glycerol Assay Kit (ab65337)
  • Free Glycerol Assay Kit - Functional Studies

Key features and details

  • Assay type: Quantitative
  • Detection method: Colorimetric/Fluorometric
  • Platform: Microplate reader
  • Assay time: 40 min
  • Sample type: Cell culture supernatant, Other biological fluids, Plasma, Serum, Tissue Extracts
  • Sensitivity: 1 µM

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Overview

  • Product name

    Free Glycerol Assay Kit
    See all Glycerol kits
  • Detection method

    Colorimetric/Fluorometric
  • Sample type

    Cell culture supernatant, Serum, Plasma, Other biological fluids, Tissue Extracts
  • Assay type

    Quantitative
  • Sensitivity

    > 1 µM
  • Range

    1 µM - 10000 µM
  • Assay time

    0h 40m
  • Product overview

    Glycerol Assay Kit (ab65337) provides a sensitive, easy assay to measure free glycerol concentration.


    In the glycerol assay protocol, glycerol is enzymatically oxidized to generate a product which reacts with the probe to generate color (λ= 570 nm) and fluorescence (Ex/Em = 535/587 nm).


    This glycerol assay kit can detect 50 pmol-10 nmol (or ~1-10000 µM range) of glycerol.


    Glycerol assay protocol summary:
    - add samples and standards to wells
    - add reaction mix and incubate for 30 min
    - analyze with a microplate reader

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called K630 Free Glycerol Colorimetric/Fluorometric Assay Kit. K630-100 is the same size as the 100 test size of ab65337.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Glycerol Assay Buffer WM 1 x 25ml
    Glycerol Enzyme Mix (lyophilized) Green 1 vial
    Glycerol Probe (in DMSO, anhydrous) Red 1 x 200µl
    Glycerol Standard (100 mM) Yellow 1 x 200µl
  • Research areas

    • Kits/ Lysates/ Other
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    • Sugar Assays
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    • Obesity
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    • Kits
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    • Lipid metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Cancer
  • Relevance

    Glycerol is widely used in foods, beverages, solvents, pharmaceutical and cosmetic products, etc. There is broad interest in quantification of glycerol for research and development.
  • Alternative names

    • Glycerin
    • Glycerine

Associated products

  • Related Products

    • Carrez Clarification Reagent Kit (ab202373)
    • Triglyceride Assay Kit - Quantification  (ab65336)

Images

  • Functional Studies - Free Glycerol Assay Kit (ab65337)
    Functional Studies - Free Glycerol Assay Kit (ab65337)

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

  • Functional Studies - Free Glycerol Assay Kit (ab65337)
    Functional Studies - Free Glycerol Assay Kit (ab65337)

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

  • Functional Studies - Free Glycerol Assay Kit (ab65337)
    Functional Studies - Free Glycerol Assay Kit (ab65337)

    Free Glycerol measured in cell culture lysates showing quantity (nmol) per 1 mln cells.

    Samples with the concentration of 1e7 cells/mL were used. Samples were diluted 2-8 fold and measured fluorometrically.

  • Functional Studies - Free Glycerol Assay Kit (ab65337)
    Functional Studies - Free Glycerol Assay Kit (ab65337)

    Free Glycerol measured in biological fluids showing quantity (nmol) per mL of tested sample. Samples were diluted 2-8 fold and measured colorimetrically.

  • Free Glycerol Assay Kit - Functional Studies
    Free Glycerol Assay Kit - Functional Studies

    a. Measurement of free glycerol in pooled human serum (10 μL). b. Measurement of free glycerol in rat liver lysate (500 μg).

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (16)

Publishing research using ab65337? Please let us know so that we can cite the reference in this datasheet.

ab65337 has been referenced in 16 publications.

  • Ishikura S  et al. The transcriptional regulator Zfat is essential for maintenance and differentiation of the adipocytes. J Cell Biochem 122:626-638 (2021). PubMed: 33522619
  • Huang M  et al. Adipose tissue lipolysis is regulated by PAQR11 via altering protein stability of phosphodiesterase 4D. Mol Metab 47:101182 (2021). PubMed: 33549845
  • Kim NY  et al. Anti-Obesity Effects of Morus alba L. and Aronia melanocarpa in a High-Fat Diet-Induced Obese C57BL/6J Mouse Model. Foods 10:N/A (2021). PubMed: 34441691
  • Raineri A  et al. 3,5-Dicaffeoylquinic Acid Lowers 3T3-L1 Mitotic Clonal Expansion and Adipocyte Differentiation by Enhancing Heme Oxygenase-1 Expression. Molecules 26:N/A (2021). PubMed: 34443613
  • Leiss V  et al. Lack of Gai2 proteins in adipocytes attenuates diet-induced obesity. Mol Metab 40:101029 (2020). PubMed: 32480042
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-10 of 10 Abreviews or Q&A

Question

Ab65337: en el protocolo de su web indica que es un kit con el que podemos detectar glicerol libre por fluorescencia a partir de sobrenadantes celulares. ¿sería posible hacerlo a partir de extractos celulares? ¿cúal sería el protocolo de lisis?
Ab1333130: en el protocolo de su web indica que es un kit cell-based para detectar glicerol por absorbancia pero también indica en el protocolo que se partiría de sobrenadantes celulares, ¿valdría para extractos celulares?

Read More

Abcam community

Verified customer

Asked on Jun 27 2013

Answer



Ab65337: Este kit puede usarse a partir de extractos celulares. Para ello, comenzar con un número aproximado de ˜2X10^6 células, suspender el pellet en 500ul (o aproximadamente 4 volúmenes) de assay buffer en hielo usando un homogeneizador (10-15 pases) hasta que se haya alcanzado una lisis eficiente, lo cual puede comprobarse al microscopio.

Centrifugar la muestra y recoger el sobrenadante para utilizarse en los ensayos posteriores. Se recomienda probar distintas diluciones de muestra para asegurar que las lecturas caen en el rango linear de la curva de estándares.

Ab1333130: En este caso no hemos probado el kit con extractos celulares, no podemos sugerir un protocolo concreto de lisis, pero pueden utilizar un protocolo estándar de extracción, similar al anterior, y optimizarlo para cada caso.

Read More

Ariana Veiga

Abcam Scientific Support

Answered on Jun 27 2013

Question

Can I use the same method of samples preparation in the protocol of Triglyceride Quantification Kit to prepare the samples to use your Free Glycerol Assay Kit to measure intracellular glycerol?

Read More

Abcam community

Verified customer

Asked on Jan 28 2013

Answer

No, I don’t think preparing samples once for both assays is a good idea. For the glycerol assay, you just need cell homogenates, whereas for the triglyceride assay you are actually preparing the lipid extracts from the cells and using them for further steps. You cannot use cell homogenates with the triglyceride assay and the lipid extracts for the glycerol assay. They are not interchangeable. So prepare samples for each assay as recommended and follow the protocol.

Read More

Abcam Scientific Support

Answered on Jan 28 2013

Question

We usually use HEPES lysis buffer to lysate cells. I want to ask can I use your Free Glycerol Assay Kit (ab65337) and Triglyceride Quantification Kit (ab65336) to measure glycerol level and triglyceride level in the cell extracts after I use this kind of lysis buffer?

Read More

Abcam community

Verified customer

Asked on Jan 25 2013

Answer

Yes, with the Free Glycerol Assay Kit, you can use HEPES for lysis, although the assay buffer provided with the kit is recommended.

With the Triglyceride Quantification Kit, we would recommend using HEPES and NP-40 since the NP-40 is important for breaking the cells.

Read More

Abcam Scientific Support

Answered on Jan 25 2013

Question

This kit is said usable with urine samples. In fact, whereas there is no problem with the standard curve (in buffer), urine samples spiked with glycerol don't give rise to any color even if spiked with glycerol at a concentration greater than the mid point of standard curve. In brief: a same concentration of glycerol diluted in buffer or in urine provides a good signal in the first case and no signal in the second one. What is the real enzymatic reaction (it is just said "glycerol is oxidised" in the data sheet)of this kit? Whatever, in the contrary of what is said on the notice, this kit does not work with urine samples.

Read More

Abcam community

Verified customer

Asked on Oct 03 2012

Answer

Thank you for your inquiry and your patience with this reply. I will be helping you further as Carolyn is out of the office.
I spoke to our development scientist about this issue and they thought that the urine contains some compound which may be blocking the assay development. The fact that the glycerol and the standards are working shows that the kit is working well.
We also have not received any complaints from the use of this kit in urine so far, so I think there should not be a problem with such sample type, albeit it depends on the way the urine was processed and stored. Can you please send us some raw data and I can consult with our scientists again in order to try and figure out what could be happening?
I look forward to your reply.

Read More

Abcam Scientific Support

Answered on Oct 03 2012

Question

Hi,



I would like to know the reaction that takes place in your glycerol kit, ab65337.

Which components are a part of it?



Best

Read More

Abcam community

Verified customer

Asked on Sep 27 2012

Answer

Thank you for your inquiry.

The exact description of the components of this kit is proprietary information.

But I am happy to confirm the way this kit works in more detail:

The probe that is used in kit ab65337is a highly sensitive and stable probe for H₂O. It is a fluorogenic (EX/EM = 535/587 nm) and Colorometric (l = 570 nm) substrate for peroxidase.
In the presence of horseradish peroxidase (HRP), the probe reacts in a 1:1 stoichiometry with H2O2 to produce highly fluorescent resorufin.

I hope this information is helpful and wish you good luck with your research.

Read More

Abcam Scientific Support

Answered on Sep 27 2012

Question

1. We would like to know the testing method or test type ?

Could you send us the instruction for use ?

2. Can this kit test pure glycerol or impure glycerol in Ester and glycerol?



We are looking forward to hearing from you soon.




Thanks & Best Regards !

Read More

Abcam community

Verified customer

Asked on May 09 2012

Answer

Thank you for your inquiry.

Please find the answers to your questions below:

1.)This Glycerol Assay Kit provides a sensitive, easy assay to measure free glycerol concentration in various samples. In the assay, glycerol is enzymatically oxidized to generate a product which reacts with the probe to generate color (λ= 570 nm) and fluorescence (Ex/Em = 535/587 nm). The kit can detect 50 pmol-10 nmol (or ˜1-10000 µM range) of glycerol in various samples.

I attached the protocol booklet for more information

2.) Free glycerol can be measured with this kit in a wide variety of samples. If I understand you correctly, you are asking for bound glycerol in tri-ester of glycerol thatpart offats and oils?

The kit most probably does not detect these since only free glycerol can be detected. But this kit has not been specifically tested with tri-esters of glycerol.

I hope this information is helpful andwish you good luck with your research.

Read More

Abcam Scientific Support

Answered on May 09 2012

Question

Hi all,
I need the msds for the free glycerol assay kit (ab65337) mentioned in the subject otherwise I cannot order this item.
Thanks.

Read More

Abcam community

Verified customer

Asked on Mar 23 2012

Answer

Thank you for contacting us.

I am happy to let you know that the Free Glycerol Assay Kitab65337does not contain any hazardous substances.

If you wish we could provide the MSDS for DMSO which would state that it is not a hazardous substance or mixture according to Regulation (EC) No. 1272/2008 ('This substance is not classified as dangerous according to Directive 67/548/EEC.')

I hope this information is helpful. Should you require the MSDS or have any further questions, please do not hesitate to contact us.

Read More

Abcam Scientific Support

Answered on Mar 23 2012

Question

Thank you for your email.   Our customer would like to receive a replacement with another lot#.   Please check this and arrange a replacement for our customer.  

Read More

Abcam community

Verified customer

Asked on Sep 27 2011

Answer

Thank you for your reply. I have contacted our Distributors Team and they will issue 1 free of charge replacement of ab65337 on your next Abcam order. I checked the lots we have in stock and this will come from a different lot but the same QC batch from the lab. We would guarantee this under the Abpromise, so do let me know if the customer has any other trouble. I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

Answered on Sep 27 2011

Question

our customer would like to know if the following kits: #AB83390, AB65342, AB83394, AB65337 can be used for cell culture. I see on your datasheet that only the Pyruvate Assay Kit can be used with biological sample such as cell culture, Do you please confirm me this?

Read More

Abcam community

Verified customer

Asked on Oct 06 2010

Answer

All of the kits: ab65337, ab65342, ab83390, ab83394 are suitable for cell and tissue culture supernatants. As well as many other biological fluids like urine, plasma and serum.

Read More

Abcam Scientific Support

Answered on Oct 06 2010

Question

I have bought the product ab65337, a kit for measuring Glycerol in cell medium. I 'm wondering how it's better to store the cell medium that is used for Glycerol measurement. Shall I store the medium at -20°C ? or at + 4°C? Thank you for this information

Read More

Abcam community

Verified customer

Asked on Feb 02 2010

Answer

Thank you for your enquiry. It would be best to use fresh sample when possible however I know that it may take time to collect all the necessary sample before running an assay. Therefore, I would recommend storing the sample at -20 °C or colder for long term, just because we do not know if any lysis has occurred or any enzymes leaked out. Generally, in an enzyme-free sample, the glycerol should be stable for a long time (6-12 months). I hope this information will be helpful. If there is anything else that I can help you with, please do not hesitate to contact me.

Read More

Abcam Scientific Support

Answered on Feb 02 2010

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