乙腦ELISA檢測(cè)試劑盒 品牌美國(guó)NOVABIOS

NOVABIOS JE Detect^TM IgM ANTIBODY

CAPTURE ELISA (MAC-ELISA)

• Positive, negative and unknown serum to be tested should be assayed in duplicate. Refer to flow chart at the end of this section for illustration of this procedure. Twenty-two test specimens can be tested in duplicate on one 96 well plate.

      乙腦ELISA檢測(cè)試劑盒 品牌美國(guó)NOVABIOS

• Mark the microtitration strips to be used.
• Dilute test sera, and controls to 1/100 using the provided Sample diluent. Use small polypropylene tubes for these dilutions and at least 4 µL of sera and positive and negative controls. For example: 4 µL serum plus 396 µL of Sample Dilution Buffer for JE Detect IgM to make 1/100 dilution.2

• Apply the 0 µL/well of 1/100 diluted test sera, JE Detect Negative Control, and JE Detect IgM Positive Control to the plate by single or multi-pipetter as appropriate. An exemplary arrangement for twenty-two test serum samples in duplicate is shown below.

Example for Serum Sample Application

• Cover the plate with parafilm just on the well opening surface, so the bottom of the plates is not covered.

Note: This is to make sure the temperature distribution is evenly spread out in all wells from bottom and sides; any extra parafilm can be cut-out once the top is sealed to block evaporation.

• Incubate the plate at 37^oC for 1hour in a humidified incubator with water container. Humidification can be achieved using a water tray at the bottom of incubator.

Note: Do not stack plates on top of each other. They should be spread out as a single layer. This is very important for even temperature distribution. Do not use CO₂ or other gases. Do not place plates in contact with any wet substances such as wet paper towels etc.

CORRECT METHOD

• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer. Use 300 µl per well in each wash cycle.

• Add 50µl /well of JERA into row A-D and 50µl /well of

NCA into row E-H by multi-pipetter.

An exemplary application for JERA and NCA is shown below.Example for JE Antigens Application

• Cover the plate with parafilm just on the well opening surface, so the bottom of the plate should not be covered (see step 5).

• Incubate the plate at 37^oC for 1 hour in a humidified incubator with a water container for humidification (see step 6).
• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer. Use 300 µl per well in each wash cycle.
• Add 50µl /well of ready to use Enzyme-HRP conjugate into all wells by multi-pipetter.
• Cover the plate with parafilm just on the well opening surface, so the bottom of the plate should not be covered (see step 5).

• Incubate the plate at 37^oC for 1hour in a humidified incubator (see step 6).

• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer.
• Add 150µl /well of EnWash into all wells by multi-pipetter.
• Incubate the plate at room temperature for 5 minutes without any cover on the plate.
• After the incubation, wash the plate 6 times with automatic plate washer using 1x Wash buffer.
• Add 75µl /well of Liquid TMB substrate into all wells 3

multi-pipetter.

• Place and incubate the plate at room temperature in a dark place (or container) for 10 minutes without any cover on the plate.
•  
• After the incubation, add 50µl /well of Stop solution into all wells by multi-pipetter and incubate at room temperature for 1 minute without any cover on the plate.
• After the incubation, read the OD 450 value with a Microplate reader.

• 更具體的說(shuō)明請(qǐng)根據(jù)以下信息

   

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  【公司名稱】 廣州健侖生物科技有限公司
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