Product nameAnti-Giantin antibody - Golgi Marker
See all Giantin primary antibodies
DescriptionRabbit polyclonal to Giantin - Golgi Marker
Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human Giantin aa 1250-1350 conjugated to keyhole limpet haemocyanin.
(Peptide available as
- This antibody gave a positive signal in cervival tissue and MCF7 whole cells using immunohistochemistry and immunocytochemistry, respectively.
Storage instructionsShipped 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 bufferpH: 7.40
Preservative: 0.02% Sodium azide
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...
PurityImmunogen affinity purified
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.
|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.|
FunctionMay participate in forming intercisternal cross-bridges of the Golgi complex.
Cellular localizationGolgi apparatus membrane.
- Information by UniProt
- 372 kDa Golgi complex associated protein antibody
- 372 kDa Golgi complex-associated protein antibody
- GCP antibody
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.
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.
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
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.
This product has been referenced in:
- 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). Read more (PubMed: 31361892) »
- 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). Read more (PubMed: 29207017) »