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Immunofluorescent 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 (PA1-061) at 2ug/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 conjugate (A27034) 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 (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||Human, Mouse, Rat|
|Published species reactivity||Yeast, Rat, Non-human primate, Hamster, Bovine, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues E(496) A G E L K P E E E I T V G P V Q K(513) of rat beta-COP.|
|Purification||Ammonium sulfate precipitation|
|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.
PA1-061 detects Golgi beta-coatomer protein (beta-COP) from mouse, human and rat tissues.
PA1-061 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects an ~110kDa protein which represents beta-COP from rat brain homogenate. Indirect immunofluorescence staining of beta-COP in NIH-3T3 cells with PA1-061 results in beta-COP localization to the Golgi complex and to distinct vesicular structures scattered throughout the cytoplasm. Golgi localization of beta-COP is sensitive to treatment with brefeldin A or ATP depletion.
The PA1-061 immunogen is a synthetic peptide corresponding to residues E(496) A G E L K P E E E I T V G P V Q K(513) of rat beta-COP. This sequence is completely conserved in human beta-COP.
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.
The deubiquitinating enzyme UBPy/USP8 interacts with TrkA and inhibits neuronal differentiation in PC12 cells.
PA1-061 was used in western blot to investigate the role USP8 on TrkA turnover in PC12 cells
|Ceriani M,Amigoni L,D'Aloia A,Berruti G,Martegani E||Experimental cell research (333:49)||2015|
The lipid raft-bound alkaline phosphatase activity increases and the level of transcripts remains unaffected in liver of merosin-deficient LAMA2dy mouse.
PA1-061 was used in western blot to study the mechanisms underlying the increased hepatic lipid raft alkaline phosphatase activity observed in a dystrophic mouse model
|Montenegro MF,Moral-Naranjo MT,Campoy FJ,Muñoz-Delgado E,Vidal CJ||Chemico-biological interactions (216:1)||2014|
Syntaxin 17 cycles between the ER and ERGIC and is required to maintain the architecture of ERGIC and Golgi.
PA1-061 was used in western blot to identify structural requirements for syntaxin 17 localization
|Muppirala M,Gupta V,Swarup G||Biology of the cell / under the auspices of the European Cell Biology Organization (103:333)||2011|
H89 sensitive kinase regulates the translocation of Sar1 onto the ER membrane through phosphorylation of ER-coupled β-tubulin.
PA1-061 was used in western blot to investigate the regulation of Sar1 translocation onto endoplasmic reticulum membrane by H89 sensitive kinase
|Nakagawa H,Miyazaki S,Abe T,Umadome H,Tanaka K,Nishimura K,Komori M,Matsuo S||The international journal of biochemistry & cell biology (43:423)||2011|
Inhibition of GTP-dependent vesicle trafficking impairs internalization of plasmalemmal eNOS and cellular nitric oxide production.
PA1-061 was used in western blot to study the effect of GTP-dependent vesicle trafficking on eNOS endocytosis and on intracellular nitric oxide production.
|Chatterjee S,Cao S,Peterson TE,Simari RD,Shah V||Journal of cell science (116:3645)||2003|
Localization of endothelial nitric-oxide synthase phosphorylated on serine 1179 and nitric oxide in Golgi and plasma membrane defines the existence of two pools of active enzyme.
PA1-061 was used in western blot to investigate the intracellular localization of phosphorylated eNOS.
|Fulton D,Fontana J,Sowa G,Gratton JP,Lin M,Li KX,Michell B,Kemp BE,Rodman D,Sessa WC||The Journal of biological chemistry (277:4277)||2002|
Differential dendritic targeting of AMPA receptor subunit mRNAs in adult rat hippocampal principal neurons and interneurons.
PA1-061 was used in immunohistochemistry to study the localization of GluA1-4 mRNAs in hippocampal principal neurons and interneurons in adult rat brain
|Cox DJ,Racca C||The Journal of comparative neurology (521:1954)||2013|
Vaccinia virus L2 protein associates with the endoplasmic reticulum near the growing edge of crescent precursors of immature virions and stabilizes a subset of viral membrane proteins.
PA1-061 was used in immunocytochemistry to investigate the role of vaccinia virus L2 protein in viral crescent formation
|Maruri-Avidal L,Weisberg AS,Moss B||Journal of virology (85:12431)||2011|
Isoform-specific localization of the deubiquitinase USP33 to the Golgi apparatus.
PA1-061 was used in immunocytochemistry to investigate the subcellular distribution of USP33 isoforms
|Thorne C,Eccles RL,Coulson JM,Urbé S,Clague MJ||Traffic (Copenhagen, Denmark) (12:1563)||2011|
Novel C-terminal motif within Sec7 domain of guanine nucleotide exchange factors regulates ADP-ribosylation factor (ARF) binding and activation.
PA1-061 was used in immunocytochemistry to characterize the structure and function of the Sec7 domain of an ADP-ribosylation factor
|Lowery J,Szul T,Seetharaman J,Jian X,Su M,Forouhar F,Xiao R,Acton TB,Montelione GT,Lin H,Wright JW,Lee E,Holloway ZG,Randazzo PA,Tong L,Sztul E||The Journal of biological chemistry (286:36898)||2011|
Intra-Golgi formation of IgM-glycosaminoglycan complexes promotes Ig deposition.
PA1-061 was used in immunocytochemistry to investigate the role of intra-Golgi IgM-glycosaminoglycan complexes in the pathogenesis of immune complex deposition disorders
|Khan SN,Cox JV,Nishimoto SK,Chen C,Fritzler MJ,Hendershot LM,Weigert M,Radic M||Journal of immunology (Baltimore, Md. : 1950) (187:3198)||2011|
TRPM7 regulates polarized cell movements.
PA1-061 was used in immunocytochemistry to investigate the role of TRPM7 in cell morphology, polarization and movements
|Su LT,Liu W,Chen HC,González-Pagán O,Habas R,Runnels LW||The Biochemical journal (434:513)||2011|
Possible involvement of calpain-like activity in normal processing of cellular prion protein.
PA1-061 was used in immunocytochemistry to investigate the normal proteolytic processing of cellular prion protein
|Hachiya N,Komata Y,Harguem S,Nishijima K,Kaneko K||Neuroscience letters (490:150)||2011|
Antibacterial autophagy occurs at PI(3)P-enriched domains of the endoplasmic reticulum and requires Rab1 GTPase.
PA1-061 was used in immunocytochemistry to investigate the mechanism for antibacterial autophagy
|Huang J,Birmingham CL,Shahnazari S,Shiu J,Zheng YT,Smith AC,Campellone KG,Heo WD,Gruenheid S,Meyer T,Welch MD,Ktistakis NT,Kim PK,Klionsky DJ,Brumell JH||Autophagy (7:17)||2011|
Resveratrol, a natural polyphenolic compound, inhibits cholera toxin-induced cyclic AMP accumulation in Vero cells.
PA1-061 was used in immunocytochemistry to investigate the effect of resveratrol on cAMP accumulation induced by cholera toxin
|Morinaga N,Yahiro K,Noda M||Toxicon : official journal of the International Society on Toxinology (56:29)||2010|
Lack of maternal Heat Shock Factor 1 results in multiple cellular and developmental defects, including mitochondrial damage and altered redox homeostasis, and leads to reduced survival of mammalian oocytes and embryos.
PA1-061 was used in immunocytochemistry to study the role of HSF1 in oogenesis
|Bierkamp C,Luxey M,Metchat A,Audouard C,Dumollard R,Christians E||Developmental biology (339:338)||2010|
Intracellular localization of Brucella abortus and Francisella tularensis in primary murine macrophages.
PA1-061 was used in immunocytochemistry to study intracellular cycles of Brucella abortus and Francisella tularensis in murine macrophages
|Celli J||Methods in molecular biology (Clifton, N.J.) (431:133)||2008|
Urocortin trafficking in cerebral microvessel endothelial cells.
PA1-061 was used in immunocytochemistry to investigate the mechanism of urocortin endocytosis in cerebral microvessel endothelial cells
|Tu H,Kastin AJ,Bjorbaek C,Pan W||Journal of molecular neuroscience : MN (31:171)||2007|
Androgen-regulated formation and degradation of gap junctions in androgen-responsive human prostate cancer cells.
PA1-061 was used in immunocytochemistry to study the gap junction formation and degradation in human prostate cancer cells.
|Mitra S,Annamalai L,Chakraborty S,Johnson K,Song XH,Batra SK,Mehta PP||Molecular biology of the cell (17:5400)||2006|
SMAP2, a novel ARF GTPase-activating protein, interacts with clathrin and clathrin assembly protein and functions on the AP-1-positive early endosome/trans-Golgi network.
PA1-061 was used in immunocytochemistry to characterize the SMAP gene family and study its function in vesicle trafficking.
|Natsume W,Tanabe K,Kon S,Yoshida N,Watanabe T,Torii T,Satake M||Molecular biology of the cell (17:2592)||2006|
Cholesterol is required for efficient endoplasmic reticulum-to-Golgi transport of secretory membrane proteins.
PA1-061 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|
CREB4, a transmembrane bZip transcription factor and potential new substrate for regulation and cleavage by S1P.
PA1-061 was used in immunocytochemistry to characterize a new transmembrane bZip transcription factor CREB4.
|Stirling J,O'hare P||Molecular biology of the cell (17:413)||2006|
Nm23H2 facilitates coat protein complex II assembly and endoplasmic reticulum export in mammalian cells.
PA1-061 was used in immunocytochemistry to investigate the role of Nm23H2 in mammalian ER export.
|Kapetanovich L,Baughman C,Lee TH||Molecular biology of the cell (16:835)||2005|
Retrograde transport of cholera toxin from the plasma membrane to the endoplasmic reticulum requires the trans-Golgi network but not the Golgi apparatus in Exo2-treated cells.
PA1-061 was used in immunocytochemistry to determine the role of the Golgi apparatus in cholera toxin trafficking by Exo2.
|Feng Y,Jadhav AP,Rodighiero C,Fujinaga Y,Kirchhausen T,Lencer WI||EMBO reports (5:596)||2004|
V region carbohydrate and antibody expression.
PA1-061 was used in immunocytochemistry to demonstrate the effect of inappropriate V region glycosylation on antibody production from expressed immunoglobulin genes.
|Gala FA,Morrison SL||Journal of immunology (Baltimore, Md. : 1950) (172:5489)||2004|
Endocytosed cation-independent mannose 6-phosphate receptor traffics via the endocytic recycling compartment en route to the trans-Golgi network and a subpopulation of late endosomes.
PA1-061 was used in immunocytochemistry to study the mechanism for the cation-independent mannose 6-phosphate receptor endocytosis.
|Lin SX,Mallet WG,Huang AY,Maxfield FR||Molecular biology of the cell (15:721)||2004|
Heparan sulfate regulates amyloid precursor protein processing by BACE1, the Alzheimer's beta-secretase.
PA1-061 was used in immunocytochemistry to study the role of heparan sulfate in amyloid precursor protein processing.
|Scholefield Z,Yates EA,Wayne G,Amour A,McDowell W,Turnbull JE||The Journal of cell biology (163:97)||2003|
Characterization of the Golgi retention motif of Rift Valley fever virus G(N) glycoprotein.
PA1-061 was used in immunocytochemistry to study the Golgi retention motif of rift valley fever virus GN glycoprotein.
|Gerrard SR,Nichol ST||Journal of virology (76:12200)||2002|
|Non-human primate||Not Cited||
Differential requirements for COPI coats in formation of replication complexes among three genera of Picornaviridae.
PA1-061 was used in immunocytochemistry to investigate the role of the COPI during the formation of replication complexes in different Picornaviridae.
|Gazina EV,Mackenzie JM,Gorrell RJ,Anderson DA||Journal of virology (76:11113)||2002|
Vesicle-associated membrane protein-associated protein-A (VAP-A) interacts with the oxysterol-binding protein to modify export from the endoplasmic reticulum.
PA1-061 was used in immunocytochemistry to study the interaction between vesicle-associated membrane protein-associated protein-A (VAP-A) and the oxysterol-binding protein and its effect on the export from the endoplasmic reticulum.
|Wyles JP,McMaster CR,Ridgway ND||The Journal of biological chemistry (277:29908)||2002|
Overexpression of an ADP-ribosylation factor-guanine nucleotide exchange factor, BIG2, uncouples brefeldin A-induced adaptor protein-1 coat dissociation and membrane tubulation.
PA1-061 was used in immunocytochemistry to investigate the role of BIG2 during the membrane association of AP-1.
|Shinotsuka C,Yoshida Y,Kawamoto K,Takatsu H,Nakayama K||The Journal of biological chemistry (277:9468)||2002|
Toll-like receptor 4 resides in the Golgi apparatus and colocalizes with internalized lipopolysaccharide in intestinal epithelial cells.
PA1-061 was used in immunocytochemistry to investigate the localization of the toll-like receptor 4 in the Golgi apparatus.
|Hornef MW,Frisan T,Vandewalle A,Normark S,Richter-Dahlfors A||The Journal of experimental medicine (195:559)||2002|
Association of a novel PDZ domain-containing peripheral Golgi protein with the Q-SNARE (Q-soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor) protein syntaxin 6.
PA1-061 was used in immunocytochemistry to investigate the association between the PDZ domain-containing peripheral Golgi protein and the Q-SNARE protein syntaxin 6.
|Charest A,Lane K,McMahon K,Housman DE||The Journal of biological chemistry (276:29456)||2001|
Retargeting of the mitochondrial protein p32/gC1Qr to a cytoplasmic compartment and the cell surface.
PA1-061 was used in immunocytochemistry to investigate the retargeting of the mitochondrial protein p32/gC1Qr in cellular compartments.
|van Leeuwen HC,O'Hare P||Journal of cell science (114:2115)||2001|
Coat proteins and vesicle budding.
PA1-061 was used in immunocytochemistry to investigate the cellular mechanism of protein trafficking
|Schekman R,Orci L||Science (New York, N.Y.) (271:1526)||1996|
|Non-human primate||Not Cited||
In vitro assembly and disassembly of coatomer.
PA1-061 was used in immunoprecipitation to study the formation and dissolution of coatomer
|Lowe M,Kreis TE||The Journal of biological chemistry (270:31364)||1995|
beta coat protein; beta-coat protein; beta-COP; coatomer protein complex, subunit beta 1; coatomer subunit beta; COPB; receptor for activated C-kinase-2
2610019B04Rik; COPB; COPB1; MSTP026; Rack2