ADAR Recombinant Rabbit Monoclonal Antibody [JU99-33]
Catalog# ET7106-72
ADAR Recombinant Rabbit Monoclonal Antibody [JU99-33]
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WB
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IF-Cell
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IHC-P
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IF-Tissue
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FC
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Human
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unconjugated
Overview
Product Name
ADAR Recombinant Rabbit Monoclonal Antibody [JU99-33]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human ADAR aa 180-280 / 1,226.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, IF-Tissue, FC
Molecular Weight
Predicted band size: 136 kDa
Positive Control
HeLa cell lysate, 293T cell lysate, SH-SY5Y cell lysate, HepG2 cell lysate, HeLa, human colon cancer tissue, human kidney tissue, human lung cancer tissue, human pancreas tissue.
Conjugation
unconjugated
Clone Number
JU99-33
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:100
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IHC-P
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1:200-1:1,000
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IF-Tissue
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1:50
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FC
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1:1,000
Target
Function
RNA-specific adenosine deaminase (ADAR1, DSH, IFI4, p136, DRADA, DSRAD, K88dsRBP) mediates RNA editing by destabilizing double stranded RNA through deamination of adenosine to inosine in structured or double-stranded RNAs. ADAR1 is expressed from an interferon-response promoter and has a Z-DNA/Z-RNA binding domain at its N-terminus. ADAR1 co-localizes with SUMO-1 in a subnucleolar region that is distinct from the fibrillar center, the dense fibrillar component and the granular component. Localization of nuclear ADAR1 is under the influence of a nucleolar localization signal (NoLS) in the middle of ADAR1 and the exporting activity of the nuclear exporter signal (NES) near the N terminus. ADAR1 upregulates nuclear factor 90 (NF90)-mediated gene expression by interacting with NF110, NF90 and NF45. ADAR1 binds short interfering RNA (siRNA), and gene silencing by siRNA is significantly more effective in mouse fibroblasts homozygous for an ADAR1 null mutation than in wild-type cells. ADAR1 may limit the efficacy of siRNA in mammalian cells.
Background References
1. Kim U et al. Molecular cloning of cDNA for double-stranded RNA adenosine deaminase, a candidate enzyme for nuclear RNA editing. Proc Natl Acad Sci U.S.A. 91:11457-11461 (1994).
2. Patterson J B et al. Expression and regulation by interferon of a double-stranded-RNA-specific adenosine deaminase from human cells: evidence for two forms of the deaminase. Mol Cell Biol 15:5376-5388 (1995).
Tissue Specificity
Ubiquitously expressed, highest levels were found in brain and lung. Isoform 5 is expressed at higher levels in astrocytomas as compared to normal brain tissue and expression increases strikingly with the severity of the tumor, being higher in the most aggressive tumors.
Post-translational Modification
Sumoylation reduces RNA-editing activity.
Subcellular Location
Cytoplasm. Nucleus.
UNIPROT
Synonyms
136 kDa double-stranded RNA-binding protein antibody
136kDa double stranded RNA binding protein antibody
Adar 1 antibody
ADAR antibody
Adar1 antibody
Adenosine deaminase acting on RNA 1 A antibody
Adenosine deaminase RNA specific 1 antibody
Adenosine deaminase RNA specific antibody
Adenosine deaminase that act on RNA antibody
AGS6 antibody
ExpandImages
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Western blot analysis of ADAR on different lysates with Rabbit anti-ADAR antibody (ET7106-72) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: SH-SY5Y cell lysate
Lane 4: HepG2 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 136 kDa
Observed band size: 150 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7106-72) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling ADAR with Rabbit anti-ADAR antibody (ET7106-72) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ADAR antibody (ET7106-72) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-ADAR antibody (ET7106-72) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-72) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-ADAR antibody (ET7106-72) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-72) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-ADAR antibody (ET7106-72) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-72) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-ADAR antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes.The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-72, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of HeLa cells labeling ADAR.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7106-72, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"