Anti-non-Gal IgM and IgG serum antibodies were evaluated using GalT-KO PBMCs and mTg Tri-KO PBMCs5 (link),19 (link) (Table S1 ). Serum Ab binding to cells was detected by FCM (FACS CANTO II, BD Biosciences). FITC-conjugated goat anti-human IgM or IgG antibody (Invitrogen) were used as described.19 (link) Data were expressed as median fluorescence intensity (MFI) “antibody index (Ab index)” which was calculated as follows: (Experimental median fluorescence intensity (MFI) – Negative control MFI)/(Positive control MFI – Negative Control MFI) × 100. Positive control was the recipient serum against Gal + pig PBMC, and negative control was secondary ab (IgM or IgG) with target cells without serum.
Protocol detail
Serum Antibody Binding Assay
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Columbia University, Nippon Medical School, Columbia University Irving Medical Center
Top 5 similar protocols
Variable analysis
independent variables
- Anti-non-Gal IgM and IgG serum antibodies
- Cell types (GalT-KO PBMCs and mTg Tri-KO PBMCs)
- Serum Ab binding to cells (detected by FCM)
- FITC-conjugated goat anti-human IgM or IgG antibody (Invitrogen)
- Positive control (recipient serum against Gal + pig PBMC)
- Negative control (secondary ab (IgM or IgG) with target cells without serum)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!