Whole-CCHFV-virion-specific IgM and IgG responses in serum were quantified by an in-house ELISA as previously described 23 (link). To measure NP or Gc specific responses we performed a sandwich ELISA. Nunc Maxisorp plates were coated with 200ng/well of mouse monoclonal antibodies against NP (clone 9D5, BEI Resources) or Gc (11E7, BEI resources). Plates were then blocked with 5% skim milk in PBS 0.05% Tween and then semi-purified whole virus lysate 23 (link) applied. Plates were washed and a 1:1000 dilution of serum applied. Bound antibodies were detected with goat horseradish peroxidase conjugated anti-monkey IgG (Seracare, catalog KPL 074–11-021) at 1:2000 and plates developed with ABTS solution (Seracare). Development was stopped with 5% sodium dodecyl sulfate in water and absorbance at 405nm read on a Synergy HTX (Biotek). For data analysis, absorbance values of wells coated with anti-NP or anti-Gc but receiving no CCHFV lysate were subtracted from wells receiving lysate. All measurements were performed in duplicate.
Abts solution
Manufactured by LGC
ABTS solution is a colorimetric reagent used in biochemical assays. It is a stable radical cation that is commonly used to measure the antioxidant capacity of samples. The ABTS solution changes color when it reacts with antioxidants, and this color change can be quantified spectrophotometrically.
Automatically generated - may contain errors
2 protocols using abts solution
1
Serological Assay for CCHFV Antibodies
2
Serological Assay for CCHFV Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole-CCHFV-virion-specific IgM and IgG responses in serum were quantified by an in-house ELISA as previously described 23 (link). To measure NP or Gc specific responses we performed a sandwich ELISA. Nunc Maxisorp plates were coated with 200ng/well of mouse monoclonal antibodies against NP (clone 9D5, BEI Resources) or Gc (11E7, BEI resources). Plates were then blocked with 5% skim milk in PBS 0.05% Tween and then semi-purified whole virus lysate 23 (link) applied. Plates were washed and a 1:1000 dilution of serum applied. Bound antibodies were detected with goat horseradish peroxidase conjugated anti-monkey IgG (Seracare, catalog KPL 074–11-021) at 1:2000 and plates developed with ABTS solution (Seracare). Development was stopped with 5% sodium dodecyl sulfate in water and absorbance at 405nm read on a Synergy HTX (Biotek). For data analysis, absorbance values of wells coated with anti-NP or anti-Gc but receiving no CCHFV lysate were subtracted from wells receiving lysate. All measurements were performed in duplicate.
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!