ET7106-72

ADAR Recombinant Rabbit Monoclonal Antibody [JU99-33]

cat.: ET7106-72

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IHC-P, IF-Tissue, FC
Clonality:	Monoclonal
Clone number:	JU99-33

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 136 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within Human ADAR aa 180-280 / 1,226.
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.
Subcellular location:	Cytoplasm. Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue FC	1:1,000 1:100 1:200-1:1,000 1:50 1:1,000
Uniprot #:	SwissProt: P55265 Human
Alternative names:	136 kDa double-stranded RNA-binding protein 136kDa double stranded RNA binding protein Adar 1 ADAR Adar1 Adenosine deaminase acting on RNA 1 A Adenosine deaminase RNA specific 1 Adenosine deaminase RNA specific Adenosine deaminase that act on RNA AGS6 AV242451 Double stranded RNA specific adenosine deaminase Double-stranded RNA-specific adenosine deaminase Double-stranded RNA-specific editase Adar DRADA Dsh Dsrad DSRAD_HUMAN dsRNA adenosine deaminase EC 3.5.4.- G1P1 IFI 4 IFI-4 IFI4 Ifi4 protein Interferon induced protein 4 Interferon inducible protein 4 Interferon-inducible protein 4 K88DSRBP mZaADAR P136 Pre-mRNA adenosine deaminase RNA adenosine deaminase 1 RNA-editing deaminase 1 RNA-editing enzyme 1

Images

Fig1: 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.

Fig2: 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.

	Fig3: 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) 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.
	Fig4: 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) 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.
	Fig5: 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) 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.

Fig6: 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.

Fig7: 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).

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