Immunofluorescent analysis of Mitofilin (green) in 3T3 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% Blocker BSA (Product # 37525) in PBS for 30 minutes at room temperature. Cells were stained with or without Mitofilin rabbit polyclonal antibody (Product # PA3-870), at a dilution of 1:100 for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:1000 for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.
|Tested species reactivity||Dog, Cat, Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant human Mitofilin.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:5000|
* 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.
|Miscellaneous PubMed (MISC)||See 1 publications below|
PA3-870 detects Mitofilin from human, mouse, rat, canine, and feline tissues.
PA3-870 has been successfully used in Immunoflorescence, and Western blotting procedures. By Western blot it detects a 90-kDa band representing PA3-870.
PA3-870 antigen is recombinant human Mitofilin.
Mitofilin is a mitochondria-associated protein expressed in all human cell types tested. Unlike coiled coil motor proteins, mitofilin is resistant to detergent extraction. The presence of mitochondrial targeting and stop-transfer sequences, along with the accessibility of mitofilin to limited proteolysis suggests that it resides predominantly in the intermembrane space, consistant with immuno-electron micrographs which show mitofilin mainly at the mitochondrial periphery.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Molecular characterization of mitofilin (HMP), a mitochondria-associated protein with predicted coiled coil and intermembrane space targeting domains.
PA3-870 was used in immunocytochemistry and western blot to characterize mitofilin at the molecular level and investigate its cellular localization
|Odgren PR,Toukatly G,Bangs PL,Gilmore R,Fey EG||Journal of cell science (109 ( Pt 9):2253)||1996|