PIERCE-15123-Reacti-Bind NeutrAvidin Coated Plates （clear, 96-well）
產品名稱：Reacti-Bind NeutrAvidin Coated Polystyrene 96-Well Plate, wi
產品貨號：PIERCE 15123
產品規(guī)格：5 PLATES
產品價格：1734
銷售直線：021-31322337、（何）
Introduction
Thermo Scientific Reacti-Bind NeutrAvidin Coated Plates are ideal for capturing biotin-labeled molecules without interference from nonspecific binding. NeutrAvidin™ Protein is deglycosylated avidin, which reduces lectin binding to undetectable levels while retaining stability and biotin-binding affinity. NeutrAvidin™ Protein offers the advantages of a near-neutral pI （6.3）, to minimize nonspecific adsorption, and the lack of the RYD sequence, which eliminates nonspecific binding to the RGD binding domain of adhesion receptors present in a variety of cells. NeutrAvidin™ Protein yields the lowest nonspecific binding among the known biotin-binding proteins. The clear, white and black plates can be used with colorimetric, chemiluminescent and fluorescent detection methods, respectively.
Example ELISA Protocol using NeutrAvidin™ Coated Plates
A.
Materials Required
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Wash Buffer: Tris-buffered saline （25 mM Tris, 150 mM NaCl; pH 7.2; Product No. 28376）, 0.1% BSA, 0.05% Tween®-20; alternatively, use Blocker™ BSA （Product No. 37520） supplemented with 0.05% Tween®-20
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Biotinylated capture antibody adjusted to 10 μg/ml, or other appropriate concentration, with Wash Buffer
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Antigen adjusted to appropriate concentration with Wash Buffer
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Primary antibody adjusted to appropriate concentration with Wash Buffer
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Enzyme-labeled secondary antibody adjusted to appropriate concentration with Wash Buffer
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Appropriate enzyme substrate: example substrates are the TMB Substrate Kit （Product No. 34021） for horseradish peroxidase and the Phosphatase Substrate Kit （Product No. 37620） for alkaline phosphatase
B.
Method
1.
Wash each well three times with 200 μl of Wash Buffer. Add 100 μl of the biotinylated capture antibody to each well and incubate for 2 hours at room temperature.
2.
Wash each well three times with 200 μl of Wash Buffer. Make a serial dilution of the antigen and add 100 μl to each well. Incubate plate for 30 minutes at room temperature.
3.
Wash each well three times with 200 μl of Wash Buffer. Add 100 μl of the primary antibody to each well and incubate plate for 30 minutes at room temperature.
4.
Wash each well three times with 200 μl of Wash Buffer. Add 100 μl of the enzyme-labeled secondary antibody to each well. Incubate plate for 30 minutes at room temperature.
5.
Wash each well three times with 200 μl of Wash Buffer.
6.
Follow the manufacturer’s instructions for the specific detection system.
Procedure for Determining Binding Activity of the NeutrAvidin™ Coated Plates
The binding activity of the plates may be tested using Biotinylated Alkaline Phosphatase （Product No. 29339） and PNPP （Product No. 37620） or Biotinylated Horseradish Peroxidase （Product No. 29139） and TMB （Product No. 34021）.
1.
Rinse each well with three times with 200 μl of wash buffer （e.g., TBS）.
2.
Prepare a 1 mg/ml solution of the biotinylated enzyme. Make 1:2 serial dilutions using a 1:1,000 dilution for the first well. Incubate the wells for 1 hour at room temperature.
3.
Wash each well three times with 200 μl of TBS containing 0.05%Tween®-20.
4.
Incubate with 100 μl of substrate solution for 15 minutes at room temperature.
5.
Measure the absorbance of each well. Active plates result in an absorbance of 0.5 to 1.0 OD at 405 nm.
