Immunofluorescence analysis of beta COP was performed using 70% confluent log phase L6 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with beta COP Rabbit Polyclonal Antibody (Product # PA1-068) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Rat|
|Published species reactivity||Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues K(494) V L A A Q E T H E G V T E D G I E(511) of rat beta-COP.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2 µg/ml|
* 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.
|Immunocytochemistry (ICC)||See 1 publications below|
PA1-068 detects coatomer-protein I beta' (COPI beta) from rat cells.
PA1-068 has been successfully used in Western blot procedures. By Western blot, this antibody detects an ~115 kDa protein representing COP beta from PC-12 cell extract. A non-specific band at ~100 kDa may be seen.
PA1-068 immunogen is a synthetic peptide corresponding to residues K(494) V L A A Q E T H E G V T E D G I E(511) of rat beta-COP. The PA1-068 immunizing peptide (Cat. # PEP-126) is available for use in neutralization and control experiments.
Coatomer proteins are involved in regulating transport between the endoplasmic reticulum (ER) and the Golgi complex and in intra-Golgi transport. There exist two coatomer-protein mechanisms (COPI and COPII) and although they have mechanistic parallels, they are molecularly distinct. The COPI coat is comprised of seven subunits (alpha-, beta-, beta'-, gamma-, delta-, epsilon-, and zeta-COP) in a complex called coatomer. Assembly of the coatomer (COPI) onto non-clathrin coated vesicles is regulated by ADP-ribosylation factor (ARF). Vesicle formation, budding, fusion, and disassembly is dependent on GDP-GTP exchange, COPI, and ARF. COPI has been shown to facilitate retrograde intracellular transport from the ER to the Golgi complex. By contrast, COPII facilitates anterograde transport between these subcellular organelles. COPII has been shown to be independently and selectively recruited to the ER relative to COPI subunits.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Uptake pathway of polyomavirus via ganglioside GD1a.
PA1-068 was used in immunocytochemistry to investigate the mechanism of entry of polyomavirus
|Gilbert J,Benjamin T||Journal of virology (78:12259)||2004|