Key features and details
- Rabbit polyclonal to GBF1
- Suitable for: WB, IP, ICC/IF, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Product nameAnti-GBF1 antibody
See all GBF1 primary antibodies
DescriptionRabbit polyclonal to GBF1
Tested applicationsSuitable for: WB, IP, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat, Guinea pig, Chimpanzee, Rhesus monkey, Chinese hamster, Orangutan
A region between residue 1809 and 1859 of Human GBF1 (NP_004184.1).
- Whole cell lysate from HeLa, 293T and NIH3T3 cells
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.09% Sodium azide
Constituents: 0.1% BSA, Tris buffered saline
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab86071 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000 - 1/10000. Predicted molecular weight: 206 kDa.|
|IP||Use at 2-5 µg/mg of lysate.|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||1/100 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionGuanine-nucleotide exchange factor (GEF) for members of the Arf family of small GTPases involved in trafficking in the early secretory pathway; its GEF activity initiates the coating of nascent vesicles via the localized generation of activated ARFs through replacement of GDP with GTP. Recruitment to cis-Golgi membranes requires membrane association of Arf-GDP and can be regulated by ARF1, ARF3, ARF4 and ARF5. Involved in the recruitment of the COPI coat complex to the endoplasmic reticulum exit sites (ERES), and the endoplasmic reticulum-Golgi intermediate (ERGIC) and cis-Golgi compartments which implicates ARF1 activation. Involved in COPI vesicle-dependent retrograde transport from the ERGIC and cis-Golgi compartments to the endoplasmatic reticulum (ER) (PubMed:16926190, PubMed:17956946, PubMed:18003980, PubMed:12047556, PubMed:12808027, PubMed:19039328, PubMed:24213530). Involved in the trans-Golgi network recruitment of GGA1, GGA2, GGA3, BIG1, BIG2, and the AP-1 adaptor protein complex related to chlathrin-dependent transport; the function requires its GEF activity (probably at least in part on ARF4 and ARF5) (PubMed:23386609). Has GEF activity towards ARF1 (PubMed:15616190). Has in vitro GEF activity towards ARF5 (By similarity). Involved in the processing of PSAP (PubMed:17666033). Required for the assembly of the Golgi apparatus (PubMed:12808027, PubMed:18003980). The AMPK-phosphorylated form is involved in Golgi disassembly during mitotis and under stress conditions (PubMed:18063581, PubMed:23418352). May be involved in the COPI vesicle-dependent recruitment of PNPLA2 to lipid droplets; however, this function is under debate (PubMed:19461073, PubMed:22185782). In neutrophils, involved in G protein-coupled receptor (GPCR)-mediated chemotaxis und superoxide production. Proposed to be recruited by phosphatidylinositol-phosphates generated upon GPCR stimulation to the leading edge where it recruits and activates ARF1, and is involved in recruitment of GIT2 and the NADPH oxidase complex (PubMed:22573891).
Sequence similaritiesContains 1 SEC7 domain.
DomainThe DCB (dimerization and cyclophiln-binding) and HUS (homology upstream of Sec7) domains are necessary for dimerization. The DCB domain is proposed to support constitutive homodimerization; the HUS domain interacts with the DCB domain which may occur intramolecular or intermolecuar.
modificationsAMPK-mediated phosphorylation at Thr-1337 is induced by 2-deoxyglucose (2-DG) and AICA ribonucleotide, and occurs during mitosis leading to membrane disassociation and inactivation of ARF1 during mitosis.
Cellular localizationGolgi apparatus, cis-Golgi network. Endoplasmic reticulum-Golgi intermediate compartment. Golgi apparatus, trans-Golgi network. Cytoplasm. Lipid droplet. Membrane. Cycles rapidly on and off early Golgi membranes (PubMed:15616190). Stabilized on membranes when complexed with ARF1-GDP and is released from both ARF1 and membranes after it catalyzes GDP displacement and ARF1 binds GTP. Continuous cycles of recruitment and dissociation of GBF1 to membranes are required for sustained ARF activation and COP I recruitment (PubMed:15813748). In neutrophils is translocated from the Golgi to the leading edge upon GPCR stimulation (PubMed:22573891). Localization to lipid droplets is questionable (PubMed:22185782).
- Information by UniProt
- ARF1GEF antibody
- BFA resistant GEF 1 antibody
- BFA-resistant GEF 1 antibody
ab86071 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab86071 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling GBF1 with ab86071 at 1/200 (1µg/ml). Detection: DAB.
All lanes : Anti-GBF1 antibody (ab86071) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg
Lane 2 : HeLa whole cell lysate at 15 µg
Lane 3 : HeLa whole cell lysate at 5 µg
Lane 4 : 293T cell lysate at 50 µg
Lane 5 : mouse NIH3T3 cell lysate at 50 µg
Developed using the ECL technique.
Predicted band size: 206 kDa
Exposure time: 10 seconds
Immunoprecipitation/ Western Blot of ab86071. Lane 1: ab86071 at 3µg/mg whole cell lysate. Lane 2: Control IgG. ab86071 at 1µg/ml for WB. Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded. Chemiluminescence with an exposure time of 3 seconds.
ab86071 staining GBF1 in HeLa cells treated with Exo-1 (ab120292), by ICC/IF. Increase in GBF1 expression correlates with increased concentration of Exo-1 as described in literature.
The cells were incubated at 37°C for 5 minutes in media containing different concentrations of ab120292 ( Exo-1) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab86071 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab86071 has been referenced in 8 publications.
- Jagdeo JM et al. N-Terminomics TAILS Identifies Host Cell Substrates of Poliovirus and Coxsackievirus B3 3C Proteinases That Modulate Virus Infection. J Virol 92:N/A (2018). WB, ICC/IF . PubMed: 29437971
- Zhao L et al. Identification of pharmacological inhibitors of conventional protein secretion. Sci Rep 8:14966 (2018). PubMed: 30297865
- Gomes-da-Silva LC et al. Photodynamic therapy with redaporfin targets the endoplasmic reticulum and Golgi apparatus. EMBO J 37:N/A (2018). PubMed: 29807932
- Vonderstein K et al. Viperin targets flavivirus virulence by inducing assembly of non-infectious capsid particles. J Virol N/A:N/A (2017). PubMed: 29046456
- Richards AL et al. Generation of unique poliovirus RNA replication organelles. MBio 5:e00833-13 (2014). ICC/IF ; Human . PubMed: 24570367
- Covington JD et al. Skeletal muscle perilipin 3 and coatomer proteins are increased following exercise and are associated with fat oxidation. PLoS One 9:e91675 (2014). WB ; Human . PubMed: 24632837
- Arita M et al. Valosin-containing protein (VCP/p97) is required for poliovirus replication and is involved in cellular protein secretion pathway in poliovirus infection. J Virol 86:5541-53 (2012). ICC/IF . PubMed: 22379090
- Arita M et al. Phosphatidylinositol 4-kinase III beta is a target of enviroxime-like compounds for antipoliovirus activity. J Virol 85:2364-72 (2011). WB ; Human . PubMed: 21177810