HA722548

NR2E1 Recombinant Rabbit Monoclonal Antibody [JE38-26]

cat.: HA722548

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE38-26

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. 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: 43 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within
Positive control:	HepG2 cell lysate, HL-60 cell lysate, SH-SY5Y cell lysate, U-87 MG cell lysate, NIH/3T3 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HL-60, mouse brain tissue, mouse colon tissue, rat brain tissue, rat colon tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000 1:100 1:1,000 1:1,000
Uniprot #:	SwissProt: Q9Y466 Human \| Q64104 Mouse Entrez Gene: 684085 Rat
Alternative names:	hTll NR2E1 / Tailless nr2e1 NR2E1_HUMAN Nuclear receptor subfamily 2 group E member 1 Nuclear receptor TLX Orphan nuclear receptor NR2E1 Protein tailless homolog Tailless homolog Tll TLX XTLL

Images

Fig1: Western blot analysis of NR2E1 on different lysates with Rabbit anti-NR2E1 antibody (HA722548) at 1/1,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: HL-60 cell lysate (20 µg/Lane)
Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
Lane 4: U-87 MG cell lysate (20 µg/Lane)
Lane 5: NIH/3T3 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (30 µg/Lane)
Lane 7: Rat brain tissue lysate (30 µg/Lane)

Predicted band size: 43 kDa
Observed band size: 50 kDa

Exposure time: 25 seconds; 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 (HA722548) 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 HL-60 cells labeling NR2E1 with Rabbit anti-NR2E1 antibody (HA722548) 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-NR2E1 antibody (HA722548) 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 mouse brain tissue with Rabbit anti-NR2E1 antibody (HA722548) 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 (HA722548) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-NR2E1 antibody (HA722548) 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 (HA722548) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-NR2E1 antibody (HA722548) 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 (HA722548) 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 rat colon tissue with Rabbit anti-NR2E1 antibody (HA722548) 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 (HA722548) 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.

Fig7: Flow cytometric analysis of HL-60 cells labeling NR2E1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722548, 1/1,000) (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).

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.