Immunofluorescent analysis of CAIN (green) in NRK cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and blocked with 3% BSA in PBS (Product # 37525) for 30 minutes at room temperature. Cells were stained with a CAIN antibody (Product # PA3-026) at a dilution of 20ug/mL in staining buffer for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (green). Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acid residues 1881-2173 of rat cain.|
|Storage buffer||whole serum diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:1,000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA3-026 detects calcineurin inhibitor (cain) from human and rat tissues.
PA3-026 has been successfully used in Western blot, immunoprecipitation, immunofluorescence, and ELISA procedures. By Western blot, this antibody detects an ~240 kDa protein representing cain from rat liver extract. Immunoprecipitation of cain from rat brain tissues shows an interaction between cain and amphiphysin 1, adaptin, and dynamin 1.
The PA3-026 immunizing peptide is a His-tagged fusion protein corresponding to amino acid residues 1881-2173 from rat cain.
Calcineurin is a widely distributed Ser/Thr, calcium and calmodulin-dependent protein phosphatase. It mediates the immunosuppressive actions of drugs such as FK506 and cyclosporin, and has also been implicated in a number of calcium-sensitive pathways in the nervous system. Calcineurin has been found to be associated with other proteins, such as calmodulin, FKBP12, the ryanodine receptor, inositol 1,4,5-trisphosphate, and a recently identified protein, calcineurin inhibitor (cain).
Cain has been shown to bind calcineurin through its C-terminal tail. Recent studies have shown cain to bind to both amphiphysin 1 and calcineurin simultaneously and therefore acts as a physiological inhibitor of calcineurin. The binding of cain to amphiphysin 1 does not affect the interaction of amphiphysin 1 with other endocytic proteins. Cain, like calcineurin, is highly expressed in neurons and other tissues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Cain, a novel physiologic protein inhibitor of calcineurin.
PA3-026 was used in western blot to characterize cain and its interaction with calcineurin as a protein inhibitor
|Lai MM,Burnett PE,Wolosker H,Blackshaw S,Snyder SH||The Journal of biological chemistry (273:18325)||1998|