Anti-DBC-1 antibody [EPR19747] (ab215852)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19747] to DBC-1
- Suitable for: IP, ICC/IF, Flow Cyt, IHC-P, WB
- Reacts with: Human
Overview
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Product name
Anti-DBC-1 antibody [EPR19747]
See all DBC-1 primary antibodies -
Description
Rabbit monoclonal [EPR19747] to DBC-1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, 293 and Jurkat whole cell lysates; Human lung cancer, breast cancer and fetal kidney lysates. IHC-P: Human colon, cervix cancer and colon cancer tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
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General notes
Previously labelled as KIAA1967.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19747 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-DBC-1 antibody [EPR19747] (ab215852) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : 293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 102 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 3 minutes; Lane 3: 10 seconds.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling DBC-1 with ab215852 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling DBC-1 with ab215852 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on human colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling DBC-1 with ab215852 at 1/700 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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DBC-1 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cells from cervix adenocarcinoma cell line) whole cell lysate with ab215852 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab215852 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution
Lane 1: HeLa whole cell lysate, 10µg (Input).
Lane 2: ab215852 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215852 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-DBC-1 antibody [EPR19747] (ab215852) at 1/1000 dilution
Lane 1 : Human lung cancer lysate
Lane 2 : Human breast cancer lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution
Predicted band size: 102 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1/2: 3 minutes; Lane 3: 3 seconds.
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Immunohistochemical analysis of paraffin-embedded human cervix cancer tissue labeling DBC-1 with ab215852 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human cervix cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling DBC-1 with ab215852 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Nuclear staining on human colon cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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