Binding of recombinant antibodies to E1 and/or E2 was determined using an ELISA in which (a) the aa 384–717 E2 ectodomain with a His6 tag or (b) a 1% triton lysate of 293T/17 cells transfected with E1E2 was captured on plates coated with Galanthus nivalis lectin (GNA) (Sigma) as described previously [30 (link)]. In brief, pre-coated plates were blocked with 1% fish skin gelatin (Sigma) and 0.05% tween in phosphate buffered saline before antibodies were added. To detect bound antibodies, the plates were incubated with horseradish peroxidase conjugated polyclonal goat anti-human IgG (Jackson, 1:2500 diluted). Bound antibodies were detected using 3,3’, 5,5’ tetramethyl benzidine (Sigma) and the reaction was stopped using H2SO4. Optical density (OD) at 450nm was measured with an EnVision Multilabel Reader (PerkinElmer).
Horseradish peroxidase conjugated polyclonal goat anti human igg
Manufactured by Jackson ImmunoResearch
Horseradish peroxidase conjugated polyclonal goat anti-human IgG is a laboratory reagent used in immunoassays and related techniques. It consists of polyclonal antibodies raised in goats against human immunoglobulin G (IgG), which have been conjugated to the enzyme horseradish peroxidase.
Automatically generated - may contain errors
2 protocols using horseradish peroxidase conjugated polyclonal goat anti human igg
1
Recombinant Antibody Binding ELISA
2
Antibody Binding Characteristics by ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To determine the binding characteristics of the antibodies, ELISA assays were performed as described previously [26] (link). In brief, plates were coated either with the HIS-tagged E2 ectodomain (Supplementary material and methods) or with Galanthus nivalis lectin on which E1E2 protein containing 293T/17 cell lysate was captured. Then, antibodies present in B cell supernatant were added and binding was detected by horseradish peroxidase-conjugated polyclonal goat antihuman IgG (Jackson). Optical density (OD) at 450nm was measured with an EnVision Multilabel Reader (PerkinElmer).
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!