Recombinant Anti-ADAR1 antibody [EPR7033] (ab126745)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR7033] to ADAR1
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ADAR1 antibody [EPR7033]
See all ADAR1 primary antibodies -
Description
Rabbit monoclonal [EPR7033] to ADAR1 -
Host species
Rabbit -
Specificity
The immunogen is designed to detect the p150 isoform and not the p110.
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Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human ADAR1 aa 200-300. The exact sequence is proprietary. The immunogen used to raise this antibody is designed to detect isoform 1 (p150) and isoforms 2-4. It does not detect Isoform 5 (p110).
Database link: P55265 -
Positive control
- WB: HEK293T, HeLa, Ramos and SH-SY5Y cell lysates. IHC-P: Human brain tissue 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. Stable for 12 months at -20°C. -
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
EPR7033 -
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
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab126745 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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WB |
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 136 kDa).
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/1000 - 1/10000. Detects a band of approximately 150 kDa (predicted molecular weight: 136 kDa). |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Function
Converts multiple adenosines to inosines and creates I/U mismatched base pairs in double-helical RNA substrates without apparent sequence specificity. Has been found to modify more frequently adenosines in AU-rich regions, probably due to the relative ease of melting A/U base pairs as compared to G/C pairs. Functions to modify viral RNA genomes and may be responsible for hypermutation of certain negative-stranded viruses. Edits the messenger RNAs for glutamate receptor (GLUR) subunits by site-selective adenosine deamination. Produces low-level editing at the GLUR-B Q/R site, but edits efficiently at the R/G site and HOTSPOT1. Binds to short interfering RNAs (siRNA) without editing them and suppresses siRNA-mediated RNA interference. Binds to ILF3/NF90 and up-regulates ILF3-mediated gene expression. -
Tissue specificity
Ubiquitously expressed, highest levels were found in brain and lung. -
Involvement in disease
Defects in ADAR are a cause of dyschromatosis symmetrical hereditaria (DSH) [MIM:127400]; also known as reticulate acropigmentation of Dohi. DSH is a pigmentary genodermatosis of autosomal dominant inheritance characterized by a mixture of hyperpigmented and hypopigmented macules distributed on the dorsal parts of the hands and feet. -
Sequence similarities
Contains 1 A to I editase domain.
Contains 2 DRADA repeats.
Contains 3 DRBM (double-stranded RNA-binding) domains. -
Post-translational
modificationsSumoylation reduces RNA-editing activity. -
Cellular localization
Cytoplasm. Nucleus > nucleolus. Isoform 1 is found predominantly in cytoplasm but appears to shuttle between the cytoplasm and nucleus. Isoform 5 is found exclusively in the nucleolus. - Information by UniProt
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Database links
- Entrez Gene: 103 Human
- Omim: 146920 Human
- SwissProt: P55265 Human
- Unigene: 12341 Human
- Unigene: 679967 Human
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Alternative names
- 136 kDa double-stranded RNA-binding protein antibody
- 136kDa double stranded RNA binding protein antibody
- Adar 1 antibody
see all
Images
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All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ADAR knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 136 kDa
Observed band size: 130 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab126745 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.
ab126745 Anti-ADAR1 antibody [EPR7033] was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADAR1 antibody [EPR7033] (ab126745)
ab126745, at 1/50 dilution, staining ADAR1 in paraffin-embedded Human brain tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of permeabilized Ramos cells, staining ADAR1 (red) with ab126745. 1x106 cells were collected and washed with blocking buffer. Cells were fixed with 2% paraformaldehyde, permeabilized with 1X FACS permeabilizing solution and blocked with blocking buffer for 30 minutes at room temperature. Cells were incubated with primary antibody (1/10) for 30 minutes at room temperature before a Fluorescently-conjugated secondary antibody or 30 min at room temperature. A rabbit IgG was used as a negative control (green).
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ADAR1 knockout HAP1 cell lysate (20 µg)
Lane 3: HepG2 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab126745 observed at 150 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab126745 was shown to recognize ADAR1 when ADAR1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging. -
All lanes : Anti-ADAR1 antibody [EPR7033] (ab126745) at 1/1000 dilution
Lane 1 : HeLa (treated with IFN-alpha) cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 136 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab126745 has been referenced in 9 publications.
- Gromisch CM et al. Humanized anti-DEspR IgG4S228P antibody increases overall survival in a pancreatic cancer stem cell-xenograft peritoneal carcinomatosis ratnu/nu model. BMC Cancer 21:407 (2021). PubMed: 33853558
- Kung CP et al. Evaluating the therapeutic potential of ADAR1 inhibition for triple-negative breast cancer. Oncogene 40:189-202 (2021). PubMed: 33110236
- Yang H et al. LINC00667 promotes the proliferation, migration, and pathological angiogenesis in non-small cell lung cancer through stabilizing VEGFA by EIF4A3. Cell Biol Int 44:1671-1680 (2020). PubMed: 32281700
- Ma C et al. Circular RNA hsa_circ_0004872 inhibits gastric cancer progression via the miR-224/Smad4/ADAR1 successive regulatory circuit. Mol Cancer 19:157 (2020). PubMed: 33172486
- Yu J et al. ADAR1 p110 Enhances Adhesion of Tumor Cells to Extracellular Matrix in Hepatocellular Carcinoma via Up-Regulating ITGA2 Expression. Med Sci Monit 25:1469-1479 (2019). PubMed: 30798327
- Lazzari E et al. Alu-dependent RNA editing of GLI1 promotes malignant regeneration in multiple myeloma. Nat Commun 8:1922 (2017). PubMed: 29203771
- Shi L et al. Circular RNA expression is suppressed by androgen receptor (AR)-regulated adenosine deaminase that acts on RNA (ADAR1) in human hepatocellular carcinoma. Cell Death Dis 8:e3171 (2017). PubMed: 29144509
- Dou N et al. Aberrant overexpression of ADAR1 promotes gastric cancer progression by activating mTOR/p70S6K signaling. Oncotarget 7:86161-86173 (2016). PubMed: 27863387
- Arias C et al. KSHV 2.0: a comprehensive annotation of the Kaposi's sarcoma-associated herpesvirus genome using next-generation sequencing reveals novel genomic and functional features. PLoS Pathog 10:e1003847 (2014). WB ; Human . PubMed: 24453964