神經醯胺酶分析試劑盒| Neuraminidase Assay Kit 貨號Abnova™ KA1633
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神經醯胺酶分析試劑盒| Neuraminidase Assay Kit 貨號Abnova™ KA1633
Neuraminidase Assay Kit 靈敏度Colorimetric Assay: 0.1 to 10 U/L, Fluorimetric Assay: 0.01 to 2 U/L
神經醯胺酶（NEURAMINDASE也稱為唾液酸酶）是一種水解末端唾液酸殘基(terminal sialic acid residues )的酶。多醣鏈(poly-saccharide chains)。它主要在細菌(bacteria )和病毒( viruses)等微生物中表達。神經醯胺酶（NEURAMINDASE也稱為唾液酸酶）對唾液酸殘基的切割被認為在流感感染中發揮多種作用病毒。它被認為有助於滲透粘膜內層、侵入靶細胞、elution來自受感染細胞的子代病毒，並防止自我聚集(self-aggregation)。因此，神經醯胺酶（NEURAMINDASE也稱為唾液酸酶）是一種流感藥物開發的重要目標以及簡單、直接和自動化的程序測量神經氨酸酶活性(neuraminidase activity)在研究和藥物發現中有著廣泛的應用。
測定原理 (Principle of the Assay)
神經氨酸酶檢測試劑盒(The Neuraminidase Assay Kit )可一步測量神經氨酸酶釋放的唾液酸(sialic acid)。變化在反應產物在 570 nm 處的顏色強度或 λem/ex = 585/530 nm 處的螢光強度可直接與樣品中的神經氨酸酶活性成正比。
The standard curve is for the purpose of illustration only and should not be used to calculate unknowns. A standard curve should be generated each time the assay is performed.
Specification
- Product Description:
- Neuraminidase Assay Kit is a quantitative colorimetric/fluorimetric determination of neuraminidase activity.
- Suitable Sample:
- Biological Sample
- Sample Volume:
- 20 uL
- Detection Method:
- Colorimetric, Fluorometric
- Assay Type:
- Quantitative
- Calibration Range:
- Colorimetric Assay: 0.1 to 10 U/L, Fluorimetric Assay: 0.01 to 2 U/L
- Regulation Status:
- For research use only (RUO)
Colorimetric Procedure
Note: SH-group containing reagents (e.g. mercaptoethanol, DTT) may interfere with this assay and should be avoided in sample preparation.
1. Equilibrate all components to desired reaction temperature (i.e 37°C). Prepare a 400 μM Standard Premix by mixing 20 μL of the 10 mM Standard and 480 μL dH2O. Dilute Standard in distilled water as follows. Transfer 20 μL standards into separate wells of a clear flat-bottom 96-well plate.
2. Transfer 20 μL of each sample into two separate wells of the same plate. One well will be used for the sample activity and one for the sample blank.
3. Immediately prior to starting the reaction, prepare enough Working Reagent (WR) for all sample and standard wells by mixing per reaction tube: 30 μL Assay Buffer, 55 μL Substrate, 1 μL Cofactors, 1 μL Enzyme and 0.5 μL Dye Reagent. For the sample blank wells, substitute 55 μL Assay Buffer for the 55 μL Substrate. Add 80 μL of the appropriate WR to each well.
4. Incubate the reaction plate protected from light at 37°C (or desired temperature) for 20 min. Measure the OD at 570 nm (OD20min). Incubate reaction plate for a further 30 min, again protected from light and at 37°C (or desired temperature). Measure the OD (OD50min)
Fluorimetric Procedure
1. Dilute the Standards prepared in Colorimetric Procedure 1:5 in H2O. Transfer 20 μL standards into
separate wells of a black 96-well plate.
2. Transfer 20 μL of each sample into two separate wells of the same plate. One well will be used for the
sample activity and one for the sample blank.
3. Add 80 μL of appropriate Working Reagent (see Colorimetric Procedure) to each well. Tap plate to mix.
4. Incubate the reaction plate protected from light at 37°C (or desired temperature) for 20 min. Measure the
F (λex/em = 530/570 nm) (F20min). Incubate reaction plate for a further 30 min, again protected from light
and at 37°C (or desired temperature). Measure the F (F50min).
