Wells of microtiter plates were coated overnight with either 100 ng M2e-tGCN4, 100 ng BM2e-tGCN4, 100 ng M2e peptide or 100 ng mouse FcγRIV protein. The coated plates were blocked with 5% milk powder in phosphate buffered saline (PBS) and dilution series of the VHHs were added to the wells. In the M2e ELISA, bound VHHs were detected with mouse anti-Histidine Tag antibody (MCA1396, Abd Serotec) followed by horseradish peroxidase (HRP)-linked anti-mouse IgG (1/2000, NXA931, GE Healthcare). In the ELISA with coated recombinant FcγRIV protein, binding was detected with a HRP conjugated rabbit anti-camelid VHH antibody (A01861-200, GenScript). After washing 50 μl of TMB substrate (Tetramethylbenzidine, BD OptETA) was added to every well. The reaction was stopped by addition of 50 μl of 1M H2SO4, after which the absorbance at 450 nM was measured with an iMark Microplate Absorbance Reader (Bio Rad).
A01861 200
Manufactured by GenScript
The A01861-200 is a lab equipment item offered by GenScript. It has a core function of providing a specific measurement or testing capability for use in laboratory settings. No further details on the intended use or performance capabilities of this product are provided.
Automatically generated - may contain errors
2 protocols using a01861 200
1
Quantification of VHH Binding to M2e and FcγRIV
2
ELISA Binding Assay for SARS and MERS Protein Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Wells of microtiter plates (type II, F96 Maxisorp, Nuc) were coated overnight at 4°C, respectively, with 100 ng recombinant MERS-CoV S-2P protein (with foldon), SARS-CoV-1 S-2P protein (with foldon), MERS-CoV RBD, MERS-CoV NTD, MERS-CoV S1, SARS-CoV-1 RBD, SARS-CoV-1 NTD or Fc-tagged SARS-CoV-2 RBD-SD1. The coated plates were blocked with 5% milk powder in PBS. Dilution series of the VHHs were added to the wells. Binding was detected by incubating the plates sequentially with either mouse anti-Histidine Tag antibody (MCA1396, Abd Serotec) followed by horseradish peroxidase (HRP)-linked anti-mouse IgG (1/2000, NXA931, GE Healthcare) or Streptavidin-HRP (554066, BD Biosciences) or by an HRP-linked rabbit anti-camelid VHH monoclonal antibody (A01861-200, GenScript). After washing 50 μL of TMB substrate (Tetramethylbenzidine, BD OptETA) was added to the plates and the reaction was stopped by addition of 50 μL of 1 M H2SO4. The absorbance at 450 nM was measured with an iMark Microplate Absorbance Reader (Bio Rad). Curve fitting was performed using nonlinear regression (Graphpad 7.0).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!