製品の概要

  • 製品名
    Pyruvate Kinase (PK) Assay Kit
  • サンプルの種類
    Cell culture supernatant, Urine, Serum, Plasma, Other biological fluids, Tissue
  • アッセイタイプ
    Quantitative
  • 検出感度
    = 0.1 mU/ml
  • 全工程の試験時間
    0h 40m
  • 製品の概要

    Pyruvate Kinase (PK) Assay Kit (ab83432) provides a simple, direct and automation-ready procedure for measuring pyruvate kinase activity in cell culture supernatant, plasma, serum, tissue, urine and other biological fluids (not validated for use with whole blood). In the assay, PEP and ADP were catalyzed by PK to generate pyruvate and ATP. The generated pyruvate is oxidized by pyruvate oxidase to produce color (at λ = 570 nm) and fluorescence (at Ex/Em = 535/587 nm). Since the increase in color or fluorescence intensity is proportional to the increase in pyruvate amount, the PK activity can be accurately measured. The kit detects 0.1 mU/ml pyruvate kinase.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    Pyruvate kinase (PK, EC 2.7.1.40) is an enzyme involved in glycolysis. It catalyzes the transfer of a phosphate group from phosphoenolpyruvate (PEP) to ADP, yielding one molecule of pyruvate and one molecule of ATP. Lack of pyruvate kinase will slow down the process of glycolysis which causes the disease known as pyruvate kinase deficiency.

  • アプリケーション
    適用あり: Functional Studiesmore details

法規制情報

製品の特性

  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 100 tests
    OxiRed Probe Red 1 x 200µl
    PK Assay Buffer WM 1 x 25ml
    PK Enzyme Mix (Lyophilised) Green 1 vial
    PK Positive Control (Lyophilised) Blue 1 vial
    PK Substrate Mix (Lyophilised) Purple 1 vial
    Pyruvate Standard (100 nmol/µl) Yellow 1 x 100µl
  • 研究分野
  • 関連性
    Pyruvate kinase (PK, EC 2.7.1.40) is an enzyme involved in glycolysis. It catalyzes the transfer of a phosphate group from phosphoenolpyruvate (PEP) to ADP, yielding one molecule of pyruvate and one molecule of ATP. Lack of pyruvate kinase will slow down the process of glycolysis which causes the disease known as pyruvate kinase deficiency.
  • 細胞内局在
    Cytoplasmic and Nuclear
  • 別名
    • CTHBP
    • EC 2.7.1.40
    • MGC3932
    • OIP3
    • OPA interacting protein 3
    • p58
    • PK
    • PK muscle type
    • PK1
    • PK2
    • PK3
    • PKL
    • PKLR
    • PKM
    • PKM2
    • PKR
    • PKRL
    • Pyruvate kinase 1
    • Pyruvate kinase 2/3
    • Pyruvate kinase isozyme R/L
    • Pyruvate kinase isozymes M1/M2
    • Pyruvate kinase liver and blood cell
    • Pyruvate kinase liver and RBC
    • Pyruvate kinase muscle
    • Pyruvate kinase type L
    • R type/L type pyruvate kinase
    • Red cell/liver pyruvate kinase
    • RPK
    • TCB
    • THBP1
    • Thyroid hormone binding protein cytosolic
    • Tumor M2 PK
    see all

アプリケーション

Our Abpromise guarantee covers the use of ab83432 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
Functional Studies Use at an assay dependent dilution.

画像

  • Pyruvate measured in biological fluid after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per ml of tested sample.

  • Pyruvate measured in mouse tissue lysates after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per mg of tested sample.
  • Pyruvate measured in cell lysates after 2 min, 10 min and 20 min incubation time showing quantity (nmol) per 1 mln of tested cells.
  • Colorimetric standard curve: mean of duplicates (+/- SD) with background reads subtracted.
  • Fluorometric standard curve: mean of duplicates (+/- SD) with background reads subtracted.

  • Pyruvate kinase samples time line using ab83432.
  • Standard curve: mean of duplicates (+/- SD) with background reads subtracted.
  • Pyruvate standard curve using ab83432.
  • PC-3 and MCF-7 cells were treated with OA at the indicated doses and PK activity was determined 12 hr after OA treatment using ab83432. The average values of three independent experiments were shown as Mean ± SD. *, P<0.05, **, P<0.01.

プロトコール

参考文献

This product has been referenced in:
  • Liu J  et al. Oleanolic acid suppresses aerobic glycolysis in cancer cells by switching pyruvate kinase type M isoforms. PLoS One 9:e91606 (2014). Functional Studies . Read more (PubMed: 24626155) »
  • Chen Z  et al. TRIM35 Interacts with pyruvate kinase isoform M2 to suppress the Warburg effect and tumorigenicity in hepatocellular carcinoma. Oncogene N/A:N/A (2014). Read more (PubMed: 25263439) »

See all 2 Publications for this product

レビューと Q&A

Thank you for contacting us.

I would not recommend to use a normal cuvette for a spectrometer/Colorimeter because the volume you will use is much much smaller (50 ul to 1 ml or more). Or you will have to use much more to product which may be...

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Thank you for contacting us.
The pyruvate kinase gene is mammalian.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us.

While we have not specifically tested ab83432 with PKM2, as long as the PKM2 isoform also catalyzes PEP and ADP to generate pyruvate and ATP, it will be detected by this kit.

The probe used in this kit i...

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Thank you for contacting us.

Unfortunately, the composition and their concentrations of each of these kit components is proprietary information, which I cannot divulge.

I am sorry for not being able to provide you any useful inform...

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Thank you for your email.

Safety datasheets (SDS) are always available from the product page on our website.

The assay buffer contains Igepal, which may cause eye damage if it comes in contact with your eyes. That is the only hazardo...

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Thank you for your patience. After reviewing your data, my colleague in the lab believes that the high background is likely caused by contamination in one or more of the reagents. Please let me know if the replacement kit is working better for you. If ...

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Thank you for your enquiry.

Theoretically the Pyruvate Kinase (PK) Assay Kit should work with isolated red blood cells (RBC). However, we have not purified RBC and tested it with this kit, therefore, we do not have any specific protocol reco...

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Thank you for contacting us.

The lab recommends incubating the standards for 20-30 minutes and using these readings to plot the standard curve. The sample readings for T1 and T2 should be such that they fall within the linear range of the sta...

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Thank you for contacting abcam regarding assay buffers.

I have had a chance to speak with the lab about this. The assay buffers of both these kits are quite different and cannot be used interchangeably.

Please let me know if you h...

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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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