Recombinant
RabMAb

Recombinant Anti-GBF1 antibody [EPR14889] - C-terminal (ab189512)

Overview

  • Product name

    Anti-GBF1 antibody [EPR14889] - C-terminal
    See all GBF1 primary antibodies
  • Description

    Rabbit monoclonal [EPR14889] to GBF1 - C-terminal
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, Flow Cyt, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human GBF1 aa 1800 to the C-terminus. The exact sequence is proprietary.
    Database link: Q92538

  • Positive control

    • HeLa, Jurkat, 293, PC12 and NIH 3T3 cell lysates; Human kidney and Mouse cardiac muscle tissues; 293 and HeLa cells.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab189512 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/250 - 1/500.
Flow Cyt 1/230.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/10000 - 1/50000. Detects a band of approximately 250 kDa (predicted molecular weight: 206 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Guanine-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).
  • Tissue specificity

    Ubiquitous.
  • Sequence similarities

    Contains 1 SEC7 domain.
  • Domain

    The 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.
  • Post-translational
    modifications

    AMPK-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 localization

    Golgi 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
  • Database links

  • Alternative names

    • ARF1GEF antibody
    • BFA resistant GEF 1 antibody
    • BFA-resistant GEF 1 antibody
    • FLJ21263 antibody
    • FLJ21500 antibody
    • GBF1 antibody
    • GBF1_HUMAN antibody
    • golgi brefeldin A resistant guanine nucleotide exchange factor 1 antibody
    • Golgi specific brefeldin A resistance factor 1 antibody
    • Golgi specific Brefeldin A-resistance factor antibody
    • Golgi-specific brefeldin A-resistance guanine nucleotide exchange antibody
    • Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 antibody
    • KIAA0248 antibody
    • MGC134877 antibody
    • MGC134878 antibody
    see all

Images

  • All lanes : Anti-GBF1 antibody [EPR14889] - C-terminal (ab189512) at 1/20000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 206 kDa
    Observed band size: 250 kDa
    why is the actual band size different from the predicted?

  • Anti-GBF1 antibody [EPR14889] - C-terminal (ab189512) at 1/50000 dilution + 293 cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 206 kDa
    Observed band size: 250 kDa why is the actual band size different from the predicted?

  • All lanes : Anti-GBF1 antibody [EPR14889] - C-terminal (ab189512) at 1/5000 dilution

    Lane 1 : PC12 cell lysate
    Lane 2 : NIH 3T3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

    Predicted band size: 206 kDa
    Observed band size: 250 kDa why is the actual band size different from the predicted?

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling GBF1 with ab189512 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin (Inset: negative control).

  • Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling GBF1 with ab189512 at 1/100 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin (Inset: negative control).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed 293 cells labeling GBF1 with ab189512 at 1/250 dilution followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary antibody at 1/200 dilution. Counter stained with DAPI (blue).

  • Flow cytometric analysis of 2% paraformaldehyde-fixed HeLa cells labeling GBF1 with ab189512 at 1/230 dilution (red) compared to a Rabbit monoclonal IgG isotype control (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.

References

ab189512 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa)
Total protein in input
4e+006 cells
Immuno-precipitation step
Protein A
Specification
HeLa

Dr. Aurelie Mousnier

Verified customer

Submitted Feb 19 2018

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