HA750243

Tyrosine Hydroxylase Recombinant Rabbit Monoclonal Antibody [SN59-03]

cat.: HA750243

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat, Cynomolgus monkey
Applications:	WB, IF-Cell, IHC-P, FC, IF-Tissue, IHC-Fr
Clonality:	Monoclonal
Clone number:	SN59-03

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 59 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within C-terminal human Tyrosine Hydroxylase.
Positive control:	Rat brain tissue lysates, PC-12 cell lysates, N2A, NIH/3T3, SH-SY5Y, mouse brain tissue, rat brain tissue, HEK-293.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P IHC-Fr FC IF-Tissue	1:1,000-1:10,000 1:50-1:200 1:2,000 1:200-1:1,000 1:1,000 1:200
Uniprot #:	SwissProt: P07101 Human \| P24529 Mouse \| P04177 Rat
Alternative names:	Dystonia 14 DYT14 DYT5b EC 1.14.16.2 OTTHUMP00000011225 OTTHUMP00000011226 ple Protein Pale TH The TY3H_HUMAN TYH Tyrosine 3 hydroxylase Tyrosine 3 monooxygenase Tyrosine 3-hydroxylase Tyrosine 3-monooxygenase Tyrosine hydroxylase

Images

	Fig1: Immunofluorescence analysis of frozen mouse brain tissue with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750243, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig2: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/2,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750243) at 1/2,000 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 rat brain tissue with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/2,000 dilution. 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 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750243) at 1/2,000 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: Western blot analysis of Tyrosine Hydroxylase on rat brain tissue lysates with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/1,000 dilution. Lysates/proteins at 20 µg/Lane. Predicted band size: 59 kDa Observed band size: 59 kDa Exposure time: 2 minutes; 10% 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 (HA750243) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
	Fig5: Western blot analysis of Tyrosine Hydroxylase on PC-12 cell lysates with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/2,000 dilution. Lysates/proteins at 15 µg/Lane. Predicted band size: 59 kDa Observed band size: 59 kDa Exposure time: 24 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 (HA750243) at 1/2,000 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.
	Fig6: Immunofluorescence analysis of paraffin-embedded mouse brain tissue labeling Tyrosine Hydroxylase with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/200 dilution. 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750243, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

	Fig7: Immunofluorescence analysis of paraffin-embedded rat brain tissue labeling Tyrosine Hydroxylase with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/200 dilution. 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA750243, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
	Fig8: Immunocytochemistry analysis of N2A cells labeling Tyrosine Hydroxylase with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution in 2% negative goat serum 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.
	Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling Tyrosine Hydroxylase with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution in 2% negative goat serum 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.

Fig10: Immunocytochemistry analysis of SH-SY5Y cells labeling Tyrosine Hydroxylase with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Tyrosine Hydroxylase antibody (HA750243) at 1/50 dilution in 2% negative goat serum 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.

Fig11: Flow cytometric analysis of HEK-293 cells labeling Tyrosine Hydroxylase.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA750243, 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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