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.
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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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