Human anti-SARS-CoV-2 nucleoprotein(NP) IgM antibody ELISA Kit

Please read this instruction manual carefully before beginning this assay.

About This Assay

It is for the qualitative determination of anti-SARS-CoV-2 N protein-IgM in human serum, plasma.

Test  Principle 

The ELISA is based on the the qualitative enzyme immunoassay technique. The microplate  provided  in  this  kit  has  been  pre-coated  with  human novel coronavirus nucleoprotein. Samples are pipetted into the wells with anti-human IgM antibody conjugated Biotin. Next, Streptavidin-HRP is added to each microplate well and incubated. After TMB Substrate solution is added, Any antibodies specific for the antigen present will bind to the pre-coated antigen. Following a wash to remove any unbound reagent, a substrate solution is added to the wells and color develops in proportion to the amount of human novel coronavirus nucleoprotein (SARS-CoV-2 NP) IgM antibody bound in the initial step. The color development is stopped and the intensity of the color is measured.

Materials Supplied

 Store  as follows for Unopened kit,  it is 12 months shelf  life. Store at 4℃ for opened kit, it is one month shelf  life.

Materials Needed But Not Supplied

1.  Microplate reader (wavelength: 450 nm)

2.  37℃  incubator

3.  Automated plate washer

4.  Precision single and multi-channel pipette and disposable tips

5.  Clean tubes and Eppendorf  tubes

6.  Deionized or distilled water

Samples Collection and Storage

Serum:  Use a serum separator tube and allow samples to clot for two hours at room temperature or overnight at 4 ℃ before centrifugation for 25 minutes at approximately   1,000×g. Assay freshly prepared serum immediately or store samples in aliquot     

at -20 ℃ or -80 ℃ for later use.  Avoid repeated freeze-thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 20 minutes at 1,000×g at 2-8 ℃ within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20 ℃ or -80 ℃ for later use. Avoid repeated freeze-thaw cycles.

Note:

1.  Samples to be used within 5 days may be stored at 4 ℃, otherwise samples must be

  stored at -20℃ (≤6 months) or -80℃ (≤12 months) to avoid loss of bioactivity and contamination.

2.  Samples hemolysis will influence the result, so hemolytic specimen should not be used.

3.  When performing the assay, bring samples to room temperature.

4. It is highly recommended to use serum instead of plasma for the detection based on

      quantity of our in-house data.

Samples Preparation

1. Dilute the serum or plasma samples with Universal Diluent(1:200) before test. The suggested 

200-fold dilution can be achieved by adding 1.5 μL sample to <s