Recombinant Anti-ATAD2 antibody [EPR12730] (ab176319)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12730] to ATAD2
- Suitable for: Flow Cyt (Intra), WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ATAD2 antibody [EPR12730]
See all ATAD2 primary antibodies -
Description
Rabbit monoclonal [EPR12730] to ATAD2 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human ATAD2 aa 50-150 (Cysteine residue). The exact sequence is proprietary.
Database link: Q6PL18 -
Positive control
- WB: HeLa, MCF-7, T47-D and Saos-2 cell lysates. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
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General notes
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12730 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab176319 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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Flow Cyt (Intra) |
1/60.
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WB |
1/1000 - 1/10000. Predicted molecular weight: 159 kDa.
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ICC/IF |
1/100 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/60. |
WB
1/1000 - 1/10000. Predicted molecular weight: 159 kDa. |
ICC/IF
1/100 - 1/250. |
Target
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Function
May be a transcriptional coactivator of the nuclear receptor ESR1 required to induce the expression of a subset of estradiol target genes, such as CCND1, MYC and E2F1. May play a role in the recruitment or occupancy of CREBBP at some ESR1 target gene promoters. May be required for histone hyperacetylation. Involved in the estrogen-induced cell proliferation and cell cycle progression of breast cancer cells. -
Tissue specificity
Highly expressed in estrogen receptor positive breast tumors and in osteosarcoma tumors. -
Sequence similarities
Belongs to the AAA ATPase family.
Contains 1 bromo domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 29028 Human
- Omim: 611941 Human
- SwissProt: Q6PL18 Human
- Unigene: 370834 Human
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Alternative names
- AAA nuclear coregulator cancer-associated protein antibody
- AAA nuclear coregulator, cancer-associated antibody
- ANCCA antibody
see all
Images
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Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: ATAD2 knockout HAP1 whole cell lysate (20 µg)Lanes 1 - 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab176319 was shown to specifically react with ATAD2 in wild type cells as signal was lost in ATAD2 knockout cells. Wild-type and ATAD2 knockout samples were subjected to SDS-PAGE. Ab176319 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 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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Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATAD2 (red) with ab176319 at a 1/60 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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Immunofluorescent staining of HeLa cells labeling ATAD2 using ab176319 at a 1/100 dilution.
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All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ATAD2 CRISPR/Cas9 edited HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 159 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab176319 was shown to react with ATAD2 in wild-type HeLa cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab264957 (CRISPR/Cas9 edited cell lysate ab257359) lane below 200kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ATAD2 CRISPR/Cas9 edited HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab176319 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution
Lane 1 : HeLa lysate
Lane 2 : MCF-7 lysate
Lane 3 : T47-D lysate
Lane 4 : Saos-2 lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 159 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab176319 has been referenced in 2 publications.
- Cao LJ et al. ATAD2 interacts with C/EBPß to promote esophageal squamous cell carcinoma metastasis via TGF-ß1/Smad3 signaling. J Exp Clin Cancer Res 40:109 (2021). PubMed: 33757572
- Wu S et al. Overexpression of microRNA-186 inhibits angiogenesis in retinoblastoma via the Hedgehog signaling pathway by targeting ATAD2. J Cell Physiol N/A:N/A (2019). PubMed: 30993715