First, regarding GPR75 unconjugated primary antibody with item number GPCR75-121AP:Whether the antibody can be used for WB, IP, and IHC experiments?
We have 2 GPCR75 antibodies (GPR75-101AP and GPR75-121AP) both antibodies are validated for Western blot application. Our GPR75-121AP is validated to work on Western blot
Second, regarding GPR75 biotinylated primary antibody with item number gpcr75.121-BIOTIN:
1. Could this antibody be used directly for WB or IHC experiments? Or is it that biotinylated primary antibodies for WB experiments do not require incubation of secondary antibodies, but require a specialized biotin kit for ECL, whereas streptavidin-labeled HRP is required for IHC?
The biotinylated GPR75-121AP is usually used in ELISA, IHC, IF or cell labeling experiments using a Strep-Avidin-HRP conjugate to localize the primary antibody in various applications.
2. Is there a recommended streptavidin magnetic bead to pull down the protein for this biotinylated primary antibody to be used in the Co-IP experiment?
We have not used the nanobeads for immunoprecipitation assay. For immunoprecipitation we used 1-1.5ug IgG and pull it down with protein A sepharose beads using 250ug total cell extracts in 250ul solution. I am sure with biotin conjugated antibody slightly less IgG will be required for pull down assays.
Finally, if this biotinylated primary antibody can be used for WB and IHC experiments, could you please provide me with a protocol?
Our standard IHC protocol required a fixed thin section on slide with antigen retrieval using Expo Buffer A in a pressure cooker for 15 minutes. Followed by blocking and primary antibody incubation (1-1.5ug IgG/ml) in humidified chambers overnight.