Overview

  • Product name

    Glucose 6 Phosphate Dehydrogenase Assay Kit (Colorimetric)
    See all Glucose 6 Phosphate Dehydrogenase kits
  • Detection method

    Colorimetric
  • Sample type

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

    Enzyme activity
  • Sensitivity

    > 0.04 mU/well
  • Assay time

    1h 00m
  • Product overview

    Glucose 6 Phosphate Dehydrogenase Assay Kit (Colorimetric) (ab102529) is a simple, sensitive and rapid assay detects the activity of Glucose 6 Phosphate Dehydrogenase (G6PDH or G6PD) in a variety of samples.


    In the G6PD assay protocol, glucose 6 phosphate is oxidized, which leads to the conversion of a nearly colorless probe to an intensely colored product with an absorbance at 450nm.


    The G6PD assay kit can detect as low as 0.04mU G6PD per well.


    G6PD assay protocol summary:
    - add samples and positive control to wells
    - add reaction mix
    - analyze on a microplate reader for 5-30 min

  • Platform

    Microplate reader

Properties

Images

  • G6PDH Activity measured in mouse tissue lysates. Protein concentration for samples varied from 7 mg/mL to 17 mg/mL. Samples were diluted 1-3 fold.

  • G6PDH Activity measured in cell lysates.

    Samples with the concentration of 4e6 cells/mL were used. Samples were undiluted.

  • G6PDH Activity measured in biological fluids. Samples were undiluted.

  • G6PDH Activity measured in mouse tissue lysates. Protein concentration for samples varied from 7 mg/mL to 17 mg/mL. Samples were undiluted.

  • G6PDH Activity measured in cell lysates.

    Samples with the concentration of 4e6 cells/mL were used. Samples were undiluted.

  • G6PDH Activity measured in biological fluids. Samples undiluted.

  • Example of positive control and pork liver samples test curves obtained using ab102529
  • Example of Standard curve obtained using ab102529

Protocols

References

This product has been referenced in:

  • Chhabra A  et al. Glucose-6-phosphate dehydrogenase is critical for suppression of cardiac hypertrophy by H2S. Cell Death Discov 4:6 (2018). Read more (PubMed: 29531803) »
  • Yamamoto Y  et al. Pentose phosphate pathway activation via HSP27 phosphorylation by ATM kinase: A putative endogenous antioxidant defense mechanism during cerebral ischemia-reperfusion. Brain Res 1687:82-94 (2018). Rat . Read more (PubMed: 29510140) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abreviews
Methanol lysis holds advantages over both NP-40 and mechanical lysis in that it much quicker, requiring only 2 minutes incubation at room temperature, followed by centrifugation and aspiration of the supernatant. Please find attached data from my experiment which shows that, despite identical NADH standard curves, lysis of human PBMCs in methanol leads to loss of measurable G6PDH activity when compared to NP-40 lysis.

Abcam user community

Verified customer

Submitted Oct 24 2014

Answer

This buffer should be fine.

We have not tested this specific combination of components in the concentrations mentioned. So there can always be some unforeseen interference, but theoretically the composition should work with this kit.

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Answer

Thank you for your enquiry.

I can confirm that plasma samples can be used with ab102529 Glucose 6 Phosphate Dehydrogenase Assay Kit.

Here is the plasma preparation protocol:

Collect whole blood into commercially available citrate-treated or heparinized tubes. Cells are removed from plasma by centrifugation for 10 minutes at 1,000-2,000 x g using a refrigerated centrifuge. Centrifugation for 15 minutes at 2,000 x g depletes platelets in the plasma sample.

The resulting supernatant is designated plasma. Following centrifugation, it is important to immediately transfer the liquid component (plasma) into a clean polypropylene tube using a Pasteur pipette. The samples should be maintained at 2-8oC while handling. If the plasma is not analyzed immediately, the plasma should be apportioned into 0.5 ml aliquots, stored, and transported at -20oC or lower. It is important to avoid freeze-thaw cycles.

Use this plasma directly with the assay kit. No homogenization or pre-treatment is required.

There is no specific volume we can recommend for the amount of plasma or for that matter any sample to be used since it is completely sample concentration and quality based. I can suggest to try a pilot experiment with several sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve.

Here are a few references in which the kit has been used which I hope may be helpful:

- Vázquez-Medina, J. et. al. Prolonged fasting increases glutathione biosynthesis in postweaned northern elephant seals. J. Exp. Biol., Apr 2011; 214: 1294 - 1299.

- I-Tung Chen, et. al. White Spot Syndrome Virus Induces Metabolic Changes Resembling the Warburg Effect in Shrimp Hemocytes in the Early Stage of Infection. J. Virol., Dec 2011; 85: 12919 - 12928.

I hope this information has been useful for you. Please do not hesitate to let me know if you have any other questions.

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Question
Answer

We have not tried using blood samples with this assay kit. Since blood is coloured, it will interfere with the colour of the reaction and contribute to the final readings. Also, since this is an enzymatic assay, you cannot use a filter to remove the proteins from blood to make it colorless, otherwise you would lose all the enzymes as well including the G6PH. I do not have a protocol which you can use for blood samples. I would instead recommend you to use serum/plasma.

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Answer

Merci de nous avoir contactés.
Le kit ab102529 (Glucose 6 Phosphate Dehydrogenase Assay Kit - Colorimetric) peut être utilisé pour des cultures cellulaires.
Il est recommandé de travailler avec 2 ou 5 millions de cellules homogénéisées avec du PBS froid ou un autre tampon de pH compris entre 6,5 et 8.
Il est recommandé de tester différentes quantités d'échantillon afin d'obtenir des valeurs dans la région linéaire.
J'ai mis en pièce jointe une image qui montre où se trouve l'onglet "Abreview" sur les fiches techniques.
J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

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Answer

I'm happy to help! Please feel free to contact me with any further questons.

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Answer

Thank you for your enquiry.

The cells can be homogenized with sonication on ice. I suggest contacting the supplier of the sonicator to confirm the best settings for cell lysis.

I would not recommend freeze/thaw but you could also use a dounce homogenizer to lyse the cells.

I hope this is helpful. Please contact us again if you have any further questions.

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Answer

For cells with this assay, 2-5 X 10^6 cells can be homogenized with an equivalent volume of ice cold PBS or other buffer (pH 6.5 - 8). Add 1 - 50µl samples into duplicate wells of a 96-well plate and bring volume to 50 µl with Assay Buffer. We suggest testing several doses of your samples to ensure readings are within the linear range.


Hope this information has been helpful for you. Please let me know if you have any other questions.

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Answer

Thank you for your patience while I waited to hear back from the lab.

Without giving up any of the proprietary information, it is a little complex to explain this reaction. Since NADH is one of the intermediates in this the reaction process utilized to produce color in this assay, we provide NADH as the standard.

I hope this information has been helpful for you and I am sorry that I cannot provide more specifics on the reactions.

Please let me know if you have any other questions.

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Answer

Thank you for contacting Abcam.


The NADH reference is correct for this kit, which is why we provide the NADH standard.

I hope this information has been helpful for you.

Please let me know if you have any other questions.

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