Immunofluorescent analysis of COP II (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a COP II polyclonal antibody (Product # PA1-069A) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 °C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a flourescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Hamster, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) T T Y L E F I Q Q N E E R D G V R (18) C of rat COPII.|
|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|
|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.
PA1-069A detects coatomer-protein II (COPII) from rat, mouse and human cells.
PA1-069A has been successfully used in Western blot and ICC/IF procedures. By Western blot, this antibody detects an ~85 kDa protein representing COPII from PC-12 cell extract.
The PA1-069A immunogen is a synthetic peptide corresponding to residues M(1) T T Y L E F I Q Q N E E R D G V R (18) C of rat COPII. This sequence is completely conserved between human, mouse, and rat. PA1-069A immunizing peptide (Cat. # PEP-127) 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. COPII consists of three parts: Sar1p and the two protein complexes, Sec23p-Sec24p and Sec13p-Sec31p.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A stress assembly that confers cell viability by preserving ERES components during amino-acid starvation.
PA1-069A was used in immunocytochemistry and western blot to study a stress assembly that supports cell viability by preserving ERES components during amino-acid starvation
|Zacharogianni M,Aguilera-Gomez A,Veenendaal T,Smout J,Rabouille C||eLife (3:null)||2014|
On and off membrane dynamics of the endoplasmic reticulum-golgi tethering factor p115 in vivo.
PA1-069A was used in immunocytochemistry to investigate the mechanism for membrane recruitment of the p115 tethering factor.
|Brandon E,Szul T,Alvarez C,Grabski R,Benjamin R,Kawai R,Sztul E||Molecular biology of the cell (17:2996)||2006|
Cholesterol is required for efficient endoplasmic reticulum-to-Golgi transport of secretory membrane proteins.
PA1-069A was used in immunocytochemistry to investigate the role of cholesterol during endoplasmic reticulum-to-Golgi transport.
|Ridsdale A,Denis M,Gougeon PY,Ngsee JK,Presley JF,Zha X||Molecular biology of the cell (17:1593)||2006|
Golgi inheritance in mammalian cells is mediated through endoplasmic reticulum export activities.
PA1-069A was used in immunocytochemistry to investigate the ER-dependent model of Golgi inheritance in mammalian cells.
|Altan-Bonnet N,Sougrat R,Liu W,Snapp EL,Ward T,Lippincott-Schwartz J||Molecular biology of the cell (17:990)||2006|
Cis-Golgi matrix proteins move directly to endoplasmic reticulum exit sites by association with tubules.
PA1-069A was used in immunocytochemistry to demonstrate the role of Golgi cis-medial matrix proteins in Golgi-to-ER traffic and in tubule formation and targeting.
|Mardones GA,Snyder CM,Howell KE||Molecular biology of the cell (17:525)||2006|
Biogenesis of tubular ER-to-Golgi transport intermediates.
PA1-069A was used in western blot to investigate the roles of intact microtubules during the formation and motility of Tubular transport intermediates.
|Simpson JC,Nilsson T,Pepperkok R||Molecular biology of the cell (17:723)||2006|
Involvement of BNIP1 in apoptosis and endoplasmic reticulum membrane fusion.
PA1-069A was used in immunoprecipitation to investigate the role of the alpha-SNAP during the apoptosis.
|Nakajima K,Hirose H,Taniguchi M,Kurashina H,Arasaki K,Nagahama M,Tani K,Yamamoto A,Tagaya M||The EMBO journal (23:3216)||2004|