antibody

Anti-Parkin Antibody [JF82-09]

ET1702-60TR

20 uL

99 USD

monoclonal

domestic rabbit

nonconjugated

JF82-09

human, mouse, rat

western blot, immunoprecipitation, flow cytometry, immunohistochemistry - paraffin section

Anti-Parkin Antibody [JF82-09]

Human,Mouse,Rat

WB,IF-Cell,IF-Tissue,IHC-P,IP,FC

Non-conjugated

Synthetic peptide within N-terminal human Parkin.

SwissProt: O60260 Human;SwissProt: Q9WVS6 Mouse;SwissProt: Q9JK66 Rat

Rabbit

JF82-09

IgG

20 uL

99 USD

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Liquid

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Protein A affinity purified.

Recombinant Rabbit monoclonal Antibody

293T cell lysates, SH-SY5Y, N2A, PC-3M, rat brain tissue, mouse testis tissue,Jurkat cell lysate,293 cell lysate.

52 kDa

Mitochondrion, mitochondrion outer membrane, endoplasmic reticulum, cytosol, Nucleus, neuron projection, postsynaptic density, presynapse.

1 mg/mL.

WB: 1:500-1:1,000 ;IF-Cell: 1:50-1:200 ;IF-Tissue: 1:50-1:200 ;IHC-P: 1:50-1:200 ;FC: 1:500-1:1,000 ;IP: Use at an assay dependent concentration.

ET1702-60_2.jpg

Immunocytochemistry analysis of SH-SY5Y cells labeling Parkin with Rabbit anti-Parkin antibody (ET1702-60) at 1/100 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-Parkin antibody (ET1702-60) at 1/100 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. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

ET1702-60_3.jpg

ICC staining of Parkin in N2A cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-60, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1702-60_4.jpg

ICC staining of Parkin in PC-3M cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-60, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

ET1702-60_5.jpg

Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Parkin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-60, 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.

ET1702-60_6.jpg

Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Parkin antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-60, 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.

ET1702-60_7.jpg

Flow cytometric analysis of SH-SY5Y cells labeling Parkin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-60, 1ug/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).

ET1702-60_8.jpg

Western blot analysis of Parkin on different lysates with Rabbit anti-Parkin antibody (ET1702-60) at 1/1,000 dilution. Lane 1: 293 cell lysate Lane 2: Jurkat cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 52 kDa Observed band size: 52 kDa Exposure time: 1 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 (ET1702-60) 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.