ET1703-97

NUR77 Recombinant Rabbit Monoclonal Antibody [JM59-11]

cat.: ET1703-97

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JM59-11

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: 64 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human NUR77 aa 10-49 / 598.
Positive control:	Rat brain tissue lysates, HepG2, Hela, NIH/3T3, human liver tissue, human colon carcinoma tissue, human breast tissue, mouse testis tissue, mouse ovarian tissue.
Subcellular location:	Cytoplasm. Nucleus. Nucleus membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC	1:2,000-1:5,000 1:50-1:200 1:50-1:200 1:50-1:1,000 1;1,000
Uniprot #:	SwissProt: P22736 Human \| P12813 Mouse \| P22829 Rat
Alternative names:	Early response protein NAK1 GFRP 1 GFRP GFRP1 Growth factor inducible nuclear protein N10 Growth Factor Inducible Nuclear Protein NP10 Growth Factor Response Protein 1 Hbr1 HMR Hormone Receptor MGC9485 N10 N10 nuclear protein NAK 1 NAK1 Nerve growth factor IB nuclear receptor variant 1 NGFIB NP 10 NP10 NR4A1 NR4A1_HUMAN Nuclear hormone receptor NUR/77 Nuclear Hormone Receptor TR3 Nuclear receptor subfamily 4 group A member 1 NUR77 NUR77, mouse, homolog of Orphan nuclear receptor HMR Orphan nuclear receptor NR4A1 Orphan nuclear receptor TR3 Orphan receptor tr3 Receptor NGFIB ST 59 ST-59 ST59 Steroid receptor TR3 Testicular receptor 3 TR 3 TR3 TR3 orphan receptor

Images

Fig1: Western blot analysis of NUR77 on different lysates with Rabbit anti-NUR77 antibody (ET1703-97) at 1/5,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: Mouse brain tissue lysate
Lane 3: Rat brain tissue lysate

Lysates/proteins at 20,40 µg/Lane.

Predicted band size: 64 kDa
Observed band size: 64 kDa

Exposure time: 3 minutes

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 (ET1703-97) at 1/5,000 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: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-NUR77 antibody (ET1703-97) at 1/5,000 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 (ET1703-97) at 1/5,000 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-NUR77 antibody (ET1703-97) at 1/5,000 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 (ET1703-97) at 1/5,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-NUR77 antibody (ET1703-97) at 1/5,000 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 (ET1703-97) at 1/5,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-NUR77 antibody (ET1703-97) at 1/5,000 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 (ET1703-97) at 1/5,000 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: Immunocytochemistry analysis of HepG2 cells labeling NUR77 with Rabbit anti-NUR77 antibody (ET1703-97) 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-NUR77 antibody (ET1703-97) 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.
	Fig7: Immunocytochemistry analysis of PC-12 cells labeling NUR77 with Rabbit anti-NUR77 antibody (ET1703-97) at 1/200 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-NUR77 antibody (ET1703-97) at 1/200 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.
	Fig8: Flow cytometric analysis of HepG2 cells labeling NUR77. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-97, 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).

Fig9: Flow cytometric analysis of PC-12 cells labeling NUR77.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1703-97, 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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