製品の概要

  • 製品名
    ENO1 Human Activity Assay Kit
  • 検出方法
    Colorimetric
  • 再現性
    Intra-Assay(同時再現性)
    サンプル N 平均値 SD CV%
    Cell extract 9 9%
    Inter-Assay(日差再現性)
    サンプル N 平均値 SD CV%
    Cell extract 4 11%
  • サンプルの種類
    Cell culture extracts, Tissue Extracts
  • アッセイタイプ
    Enzyme activity
  • 検出感度
    125 ng/ml
  • 検出範囲
    125 ng/ml - 2000 ng/ml
  • 添加回収試験

    90 %

    特定サンプルでの回収試験
    サンプルの種類 平均 % 測定範囲
    Cell culture supernatant 63 % - %
    Serum 140 % - %

  • 種交差性
    交差種: Human
    非交差種: Mouse, Rat
  • 製品の概要

    ab117994 is used to determine enolase 1 (ENO1) activity in a sample. The native enzyme is immunocaptured within the wells of the microplate; this removes all other enzymes. The enolase activity is determined via a coupled reaction to the consumption of NADH in an assay buffer.

  • 試験プラットフォーム
    Microplate reader

製品の特性

画像

  • Analysis of recombinant ENO1 (ab89248) and the neuron-specific recombinant ENO2 (ab78797), each at 2000 ng/mL. The relative ENO activity was determined from HepG2 standard curve as described in the protocol. The data show that the activity of the neuron-specific ENO2 can’t be detected.
  • Example of dilution series prepared from HepG2 cells extract.
  • Immunofluorescent detection of ENO1 in cytosol in HeLa cells using ab112994.
  • Scheme of reactions used in this kit. Abbreviations are as follows: enolase (ENO), 2-phospho-D-glycerate (2PG), phosphoenolpyruvate (PEP), pyruvate kinase (PK), lactate dehydrogenase (LDH).

プロトコール

参考文献

This product has been referenced in:
  • Marchesini MI  et al. A T4SS Effector Targets Host Cell Alpha-Enolase Contributing to Brucella abortus Intracellular Lifestyle. Front Cell Infect Microbiol 6:153 (2016). Read more (PubMed: 27900285) »
  • Bottermann K  et al. Systematic Analysis Reveals Elongation Factor 2 and a-Enolase as Novel Interaction Partners of AKT2. PLoS One 8:e66045 (2013). Read more (PubMed: 23823123) »
See all 2 Publications for this product

レビューと Q&A

1-2 of 2 Abreviews or Q&A

Answer

Regarding LDH control:
Please note that the LDH is simply added to the rehydrated 'Reagent Mix', along with PK, to produce the 1x Acitvity Solution.

Regarding data analysis:
If data collected in a kinetic mode, they are presented as a change of absorbance at 340 nm per unit of time, for example delta mOD/min, where mOD= 0.001 OD, as in the protocol booklet. The ENO1 activity of unknown samples can be expressed in relative arbitrary units, this is sufficient for most of the purposes. For this, follow the steps below:


1. Since consumed NADH is measured, the delta mOD/min is a negative number. So for convenience, convert the collected data to positive numbers by multiplying them by -1.
2. Subtract “zero standard” – see step 8.3 - from all other readings.
3. Construct standard curve of the reference sample dilution. As recommended, the standard Eno1 protein (ab89248) should be used as a reference sample, but other samples may be used as reference sample. Plot the protein concentration of the standard on the X axis and the corresponding zero standard-subtracted mOD/min values on Y axis as in Figure 2.. Construct a best fit curve through the datapoints. 4 parameter logistic (4PL) algoritm provides usually the best fit, though other algoritms may be used.
4. Relative activity of the unknown samples, e.g cell extract is then interpolated from the standard curve by using zero standard- subtracted mOD/min values of unknown samples as parameters on Y axis and from the standard curve interpolating the relative activities on the X axis.
5. As written in the protocol, the relative activity is the amount of reference sample required to generate the same amount of activity as the unknown sample.
6. Divide the relative activity of unknown samples by the total protein of the sample. This will allow to compare unknown samples one to another. This step is important only if unknown samples of different protein concentrations were analyzed.

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Answer

Vielen Dank für Ihren Anruf.

Bitte entschuldigen Sie die verspätete Antwort. MitoSciences sitzt an der Westküste in den USA und durch die Zeitverschiebung habe ich erst jetzt eine Antwort erhalten.

Ich freue mich Ihnen bestätigen zu können, dass in Kit ab117994 Nunc Platten (cat # 445101) namens C8 Maxisorp Nunc Immuno Module verwendet werden.

Ich hoffe, diese Information ist hilfreich und dass Ihr Analyzer diese Sorte Platten erkennt.

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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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