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    giantin-antibody-golgi-marker-ab80864.pdf

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Tags & Cell Markers Subcellular Markers Organelles Golgi
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Validated using a knockout cell line

Anti-Giantin antibody - Golgi Marker (ab80864)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (14)

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Western blot - Anti-Giantin antibody - Golgi Marker (ab80864)
  • Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Giantin antibody - Golgi Marker (ab80864)
  • Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)

Key features and details

  • Rabbit polyclonal to Giantin - Golgi Marker
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

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Protein
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Anti-Pericentrin antibody [EPR21987] (ab220784)

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Overview

  • Product name

    Anti-Giantin antibody - Golgi Marker
    See all Giantin primary antibodies
  • Description

    Rabbit polyclonal to Giantin - Golgi Marker
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to Human Giantin aa 1250-1350 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab101412)

  • Positive control

    • This antibody gave a positive signal in cervival tissue and MCF7 whole cells using immunohistochemistry and immunocytochemistry, respectively.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • Golgi

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Giantin protein (ab152432)
  • Related Products

    • Golgi Staining Kit - Green Fluorescence - Cytopainter (ab139483)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab80864 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
IHC-P (1)
Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF (1)
Use a concentration of 1 µg/ml.
WB
1/1000. Predicted molecular weight: 376 kDa.
Notes
IHC-P
Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.
WB
1/1000. Predicted molecular weight: 376 kDa.

Target

  • Function

    May participate in forming intercisternal cross-bridges of the Golgi complex.
  • Cellular localization

    Golgi apparatus membrane.
  • Target information above from: UniProt accession Q14789 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2804 Human
    • Omim: 602500 Human
    • SwissProt: Q14789 Human
    • Unigene: 213389 Human
    • Alternative names

      • 372 kDa Golgi complex associated protein antibody
      • 372 kDa Golgi complex-associated protein antibody
      • GCP antibody
      • GCP372 antibody
      • Giantin antibody
      • GOGB1_HUMAN antibody
      • GOLGB1 antibody
      • Golgi autoantigen golgin subfamily a member 1 antibody
      • Golgi autoantigen golgin subfamily b macrogolgin (with transmembrane signal) 1 antibody
      • Golgi autoantigen, golgin subfamily b, macrogolgin 1 antibody
      • Golgi complex associated protein 372kDa antibody
      • Golgi complex-associated protein 372kDa antibody
      • Golgin B1 antibody
      • Golgin B1, golgi integral membrane protein antibody
      • Golgin subfamily B member 1 antibody
      • GOLIM1 antibody
      • Macrogolgin antibody
      • MGC33154 antibody
      see all

    Images

    • Western blot - Anti-Giantin antibody - Golgi Marker (ab80864)
      Western blot - Anti-Giantin antibody - Golgi Marker (ab80864)
      All lanes : Anti-Giantin antibody - Golgi Marker (ab80864) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : GOLGB1 knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 376 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab80864 observed at 377kDa. Red - loading control, ab130007, observed at 130kDa.

      ab80864 was shown to recognize GOLGB1 in wild-type HAP1 cells as signal was lost at the expected MW in GOLGB1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GOLGB1 knockout samples were subjected to SDS-PAGE. Ab80864 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)
      Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)

      ab80864 stained in Hela cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab80864 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were ab150120 (pseudo-colored red) and ab150081 (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Giantin antibody - Golgi Marker (ab80864)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Giantin antibody - Golgi Marker (ab80864)

      IHC image of ab80864 staining in human cervix carcinoma formalin fixed paraffin embedded tissue section*, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab80864, 0.1µg/ml for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)
      Immunocytochemistry/ Immunofluorescence - Anti-Giantin antibody - Golgi Marker (ab80864)This image is courtesy of an anonymous Abreview

      Ab80864 staining Giantin in WIF-B cells (hepatoma-derived hybrid cell line) by ICC/IF (Immunocytochemistry/Immunofluorescence). Cells were fixed with formaldehyde; permeabilized with 0.2% Triton X-100 and blocked with 1% Donkey serum in 0.1% PBST for 1 hour at 22°C. Samples were incubated at 1/25 dilution for 3 hours at 22°C. An Alexa Fluor® 594 Donkey anti rabbit was used as the secondary antibody at 1/200 dilution.

      See Abreview

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (14)

    Publishing research using ab80864? Please let us know so that we can cite the reference in this datasheet.

    ab80864 has been referenced in 14 publications.

    • Wang Y  et al. Generation of a caged lentiviral vector through an unnatural amino acid for photo-switchable transduction. Nucleic Acids Res N/A:N/A (2019). PubMed: 31361892
    • Yarur HE  et al. Type 2ß Corticotrophin Releasing Factor Receptor Forms a Heteromeric Complex With Dopamine D1 Receptor in Living Cells. Front Pharmacol 10:1501 (2019). PubMed: 31969820
    • Ishida M & Bonifacino JS ARFRP1 functions upstream of ARL1 and ARL5 to coordinate recruitment of distinct tethering factors to the trans-Golgi network. J Cell Biol 218:3681-3696 (2019). PubMed: 31575603
    • Bachelot A  et al. A common African variant of human connexin 37 is associated with Caucasian primary ovarian insufficiency and has a deleterious effect in vitro. Int J Mol Med 41:640-648 (2018). PubMed: 29207017
    • Tian S  et al. Genome-wide CRISPR screens for Shiga toxins and ricin reveal Golgi proteins critical for glycosylation. PLoS Biol 16:e2006951 (2018). PubMed: 30481169
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Giantin antibody - Golgi Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (cervix)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
    Permeabilization
    Yes - Commercial Wash Buffer with Tween
    Specification
    cervix
    Fixative
    Paraformaldehyde
    Read More

    Mr. Rudolf Jung

    Verified customer

    Submitted Apr 14 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Giantin antibody - Golgi Marker

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (WIF-B cells, the hepatoma-derived hybrid cell line)
    Permeabilization
    Yes - 0.2% Triton X-100
    Specification
    WIF-B cells, the hepatoma-derived hybrid cell line
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 17 2016

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