Protocol detail

Generation of Rat and Rabbit Antibodies to VAPB and PTPIP51

Find Similar Protocols

Rat and rabbit antibodies to VAPB and PTPIP51 were generated by immunisation with GST-VAPB(1-220) and GST-PTPIP51(36-470) respectively^{20 (link)}. Rabbit anti-PTPIP51 was from Atlas, rabbit anti-hemagglutin (HA), mouse anti-α-tubulin (DM1A) and rabbit anti-Mitofusin2 were from Sigma. Rabbit anti-TOM20 was from Santa Cruz Biotechnology. Rabbit anti-GFP and mouse anti-βactin were from Abcam. Mouse PDI (RL77) was from Affinity Bioreagents. Rabbit anti-TDP-43 (12892-1-AP) was from Proteintech. Antibodies to total and ser9 phosphorylated (inactive) GSK3β were from BD Transduction Labs (mouse 610201) and Cell Signalling (rabbit 9336) respectively. The concentrations used for the different antibodies are shown in Supplementary Table 1. Secondary antibodies were horseradish peroxidase-coupled goat anti-mouse, anti-rabbit and anti-rat Igs (GE Healthcare), and Alexa fluorophore (350, 488 and 568) coupled goat anti-mouse and anti-rabbit IgGs (Invitrogen). AR-A014418 and CT99021 were from Abcam and Cayman respectively.

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.

Stoica R., De Vos K.J., Paillusson S., Mueller S., Sancho R.M., Lau K.F., Vizcay-Barrena G., Lin W.L., Xu Y.F., Lewis J., Dickson D.W., Petrucelli L., Mitchell J.C., Shaw C.E, & Miller C.C. (2014). ER–mitochondria associations are regulated by the VAPB–PTPIP51 interaction and are disrupted by ALS/FTD-associated TDP-43. Nature Communications, 5, 3996.

Publication 2014

mouse anti-α-tubulin (DM1A) rabbit anti-Mitofusin2 Rabbit anti-TOM20 Rabbit anti-GFP and mouse anti-βactin Mouse PDI (RL77) Rabbit anti-TDP-43 (12892-1-AP) Antibodies to total and ser9 phosphorylated (inactive) GSK3β mouse 610201) AR-A014418 CT99021

Anti iggs Antibodies Ar a014418 Cayman Ct99021 Goat Gsk3β Horseradish peroxidase Immunisation Mitofusin2 Mouse Rabbit Tdp 43 α tubulin

Top 5 similar protocols

Variable analysis

independent variables

Immunization with GST-VAPB(1-220) to generate rat and rabbit antibodies to VAPB
Immunization with GST-PTPIP51(36-470) to generate rat and rabbit antibodies to PTPIP51

dependent variables

The generated antibodies to VAPB and PTPIP51

control variables

The use of GST-tagged proteins as immunogens to generate the antibodies

controls

Positive controls: None specified
Negative controls: None specified

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.

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?

Revolutionizing how scientists
search and build protocols!