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
Overview
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Product name
Anti-Giantin antibody - Golgi Marker
See all Giantin primary antibodies -
Description
Rabbit polyclonal to Giantin - Golgi Marker -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, ICC/IF, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human Giantin aa 1250-1350 conjugated to keyhole limpet haemocyanin.
(Peptide available asab101412) -
Positive control
- ICC/IF: MCF7 cells. IHC-P: Human kidney tissue.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
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.
Application | Abreviews | Notes |
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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.
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ICC/IF | (1) |
Use a concentration of 1 µg/ml.
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WB |
1/1000. Predicted molecular weight: 376 kDa.
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Notes |
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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
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Function
May participate in forming intercisternal cross-bridges of the Golgi complex. -
Cellular localization
Golgi apparatus membrane. - Information by UniProt
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Database links
- Entrez Gene: 2804 Human
- Omim: 602500 Human
- SwissProt: Q14789 Human
- Unigene: 213389 Human
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Alternative names
- 372 kDa Golgi complex associated protein antibody
- 372 kDa Golgi complex-associated protein antibody
- GCP antibody
see all
Images
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ab80864 staining Giantin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab80864 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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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 kDaLanes 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.
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IHC image of ab80864 staining in human kidney 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
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (24)
ab80864 has been referenced in 24 publications.
- Zhu Y et al. Allosteric inhibition of SHP2 uncovers aberrant TLR7 trafficking in aggravating psoriasis. EMBO Mol Med 14:e14455 (2022). PubMed: 34936223
- Xu X et al. Specific motifs mediate post-synaptic and surface transport of G protein-coupled receptors. iScience 25:103643 (2022). PubMed: 35024582
- Liliac IM et al. E-Cadherin Modulation and Inter-Cellular Trafficking in Tubular Gastric Adenocarcinoma: A High-Resolution Microscopy Pilot Study. Biomedicines 10:N/A (2022). PubMed: 35203558
- Luo Q et al. Nondegradable ubiquitinated ATG9A organizes Golgi integrity and dynamics upon stresses. Cell Rep 40:111195 (2022). PubMed: 35977480
- Elmasri Z et al. Requirement of a functional ion channel for Sindbis virus glycoprotein transport, CPV-II formation, and efficient virus budding. PLoS Pathog 18:e1010892 (2022). PubMed: 36191050