製品の概要

  • 製品名
    Hexokinase Activity Assay Kit (Colorimetric)
    Hexokinase キット 製品一覧
  • 検出方法
    Colorimetric
  • サンプルの種類
    Serum, Tissue, Adherent cells, Suspension cells, Tissue Extracts
  • アッセイタイプ
    Quantitative
  • 検出感度
    < 0.1 mU/well
  • 全工程の試験時間
    1h 00m
  • 種交差性
    交差種: Mammals, Other species
  • 製品の概要

    Hexokinase Activity Assay Kit (ab136957) is a product where glucose is converted to glucose-6-phosphate by hexokinase; the glucose-6-phosphate is oxidized by glucose-6-phosphate dehydrogenase to form NADH, which reduces a colorless probe to a colored product with strong absorbance at 450 nm.


    This kit has a simple, sensitive and rapid procedure which can detect hexokinase activity even less than 0.1 mU/well.

  • 特記事項

    Hexokinases (HK) are found in many organisms including bacteria, plants and mammals and play an important role in glucose metabolism. Hexokinases phosphorylate glucose and generate glucose-6-phosphate for glycolysis. Hexokinases have four isoforms (HK-I, II, III and IV). HK-I, HK-II and HK-III have low Km, while HK-IV has 100 fold high Km. Hexokinase deficiency leads to severe human diseases such as X-linked muscular dystrophy and a rare autosomal recessive hemolytic anemia. On the other hand, increased hexokinase activity is detected in various human tumors and is associated with metastasis. Early detection of abnormal hexokinase activity is crucial for diagnosis, prediction and treatment of the disease.

  • 試験プラットフォーム
    Microplate reader

製品の特性

画像

  • Hexokinase Activity measured in cell lysates showing quantity (nmol) per 1 mln cells after 35 min of incubation.

    Samples with the concentration of 5e6 cells/mL were used. Samples were diluted 1-27 fold.

  • Hexokinase Activity measured in tissue lysates showing quantity (nmol) per mg of extracted protein after 35 min of incubation.

    Protein concentration for samples varied from 13 mg/mL to 43 mg/mL. Samples were diluted 1-9 fold.

  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted
  • NADH standard curve (a). Hexokinase activity in Positive Control and Jurkat cell lysate (40 µg) (b).

プロトコール

参考文献

This product has been referenced in:
  • Yao J  et al. By blocking hexokinase-2 phosphorylation, limonin suppresses tumor glycolysis and induces cell apoptosis in hepatocellular carcinoma. Onco Targets Ther 11:3793-3803 (2018). Read more (PubMed: 30013360) »
  • Bramer SA  et al. Hexokinase 2 drives glycogen accumulation in equine endometrium at day 12 of diestrus and pregnancy. Reprod Biol Endocrinol 15:4 (2017). Functional Studies ; Horse . Read more (PubMed: 28056994) »
See all 4 Publications for this product

レビューと Q&A

1-9 of 9 Abreviews or Q&A

Answer



When preparing these samples please make sure to keep these on ice throughout the preparation procedure. Fresh samples will give you the best results with this kit, however, frozen samples can also be used. If freezing your samples, make sure to snap-freeze them at -80ºC in order to preserve them before using them with this kit.

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Answer

We have not specifically tested the kit with plasma sample however, it should work with mammalian samples including plasma. It should not affect the sensitivity or accuracy of the test.

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Answer

This kit is based on function of the Hexokinase enzyme (conversion of glucose to glucose6phosphate is measured) so this kit does not distinguish between the isoforms.

HKI, HKII and HKIII have low Km, while HKIV has 100 fold high Km. Hence it is very likely that you will measure hexokinase activity from I, II and III and small contribution if any from HK IV depending on the relative amounts of these isoforms in your sample.

HK-II is the predominant form in adipose and muscle cells. HKII is insulin-sensitive. So if the samples are from adipose tissue or muscle the customer will measure HK-II activity. Our positive control is HK-II from Bacillus sp.

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Answer

I can confirm the the Hexokinase substrate kit component included in the kit is glucose. This component was recently re-labeled. The error in the naming will be fixed shortly and a new protocol will be available for download from the website shortly. Unfortunately, the concentration of these components is proprietary information.

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Question
Answer

Yes, this buffer contains NP-40.

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Question
Answer

The assay buffer contains NP-40.

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Answer

I apologize for any misunderstanding regarding the unit label for this kit. I am glad you read through the documentation. You are correct in your assessment of how many actual samples can be tested using this kit, after taking into account controls and blanks.

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Answer

Unfortunately, the concentration of these components is proprietary information.

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Answer

Thank you for contacting us.

I have spoken with the lab in regards to these issues. While the positive control information is proprietary, I can confirm that this is a protein control.

Regarding separate purchase of the kit's developer, I am sorry but this cannot be offered a this time. I do apologize for the inconvenience.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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