WB
Western blot analysis of Parkin on different lysates with Rabbit anti-Parkin antibody at 1/2,000 dilution. Exposure time: 6 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Parkin antibody at 1/1,000 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 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.IHC
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Parkin antibody at 1/1,000 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 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.ICC/IF
Immunocytochemistry analysis of SH-SY5Y cells labeling Parkin with Rabbit anti-Parkin antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Parkin antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of SH-SY5Y cells labeling Parkin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Parkin was immunoprecipitated from 0.2 mg SH-SY5Y cell lysate with Parkin Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Mouse anti-Parkin antibody at 1/1,000 dilution. Anti-Mouse IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SH-SY5Y cell lysate (input). Lane 2: Parkin Recombinant Rabbit Monoclonal Antibody IP in SH-SY5Y cell lysate. Lane 3: Mouse IgG instead of Parkin Recombinant Rabbit Monoclonal Antibody in SH-SY5Y cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 2 minutes.IF-P
Application: Immunofluorescence (IF-tissue). Species: Mouse. Tissue: Hippocampus. Sample: Paraffin-embedded section. Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST. Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: Parkin Recombinant Rabbit Monoclonal Antibody, 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.IF-P
Application: Immunofluorescence (IF-tissue). Species: Rat. Tissue: Brain. Sample: Paraffin-embedded section. Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: Parkin Recombinant Rabbit Monoclonal Antibody, 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.IF-F
Application: IHC-Fr. Species: Mouse. Site: Cerebral cortex. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.IF-F
Application: IHC-Fr. Species: Rat. Site: Cerebral cortex. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.| Product Name | Parkin Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human Parkin aa 1-200. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIF-FIF-PIHCIPWB |
| WB:1:2000-1:10000 IHC:1:1000 ICC/IF:1:50-1:100 IF-P:1:200 IF-F:1:500 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | PRKN |
|---|---|
| Gene Synonyms | PDJ AR-JP LPRS2 PARK2 |
| Gene Full Name | parkin RBR E3 ubiquitin protein ligase |
| Gene Summary | The precise function of this gene is unknown; however, the encoded protein is a component of a multiprotein E3 ubiquitin ligase complex that mediates the targeting of substrate proteins for proteasomal degradation. Mutations in this gene are known to cause Parkinson disease and autosomal recessive juvenile Parkinson disease. Alternative splicing of this gene produces multiple transcript variants encoding distinct isoforms. Additional splice variants of this gene have been described but currently lack transcript support. [provided by RefSeq, Jul 2008] |
| Alternative Names | AR JP antibody E3 ubiquitin ligase antibody E3 ubiquitin protein ligase parkin antibody E3 ubiquitin-protein ligase parkin antibody FRA6E antibody LPRS 2 antibody LPRS2 antibody PARK 2 antibody Park2 antibody Parkin 2 antibody Parkinson disease (autosomal recessive juvenile) 2 antibody Parkinson disease (autosomal recessive juvenile) 2 parkin antibody Parkinson disease protein 2 antibody Parkinson juvenile disease protein 2 antibody Parkinson protein 2 E3 ubiquitin protein ligase antibody Parkinson protein 2 E3 ubiquitin protein ligase (parkin) antibody PDJ antibody PRKN 2 antibody PRKN antibody PRKN2 antibody PRKN2_HUMAN antibody Ubiquitin E3 ligase PRKN antibody |
| Molecular Weight(MW) | 52 kDa |
| Cellular Localization | Mitochondrion, mitochondrion outer membrane, endoplasmic reticulum, cytosol, Nucleus, neuron projection, postsynaptic density, presynapse. |
WB
Western blot analysis of Parkin on different lysates with Rabbit anti-Parkin antibody at 1/2,000 dilution. Exposure time: 6 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Parkin antibody at 1/1,000 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 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Parkin antibody at 1/1,000 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 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.
ICC/IF
Immunocytochemistry analysis of SH-SY5Y cells labeling Parkin with Rabbit anti-Parkin antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Parkin antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of SH-SY5Y cells labeling Parkin. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Parkin was immunoprecipitated from 0.2 mg SH-SY5Y cell lysate with Parkin Recombinant Rabbit Monoclonal Antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Mouse anti-Parkin antibody at 1/1,000 dilution. Anti-Mouse IgG for IP, VHH(HRP) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: SH-SY5Y cell lysate (input). Lane 2: Parkin Recombinant Rabbit Monoclonal Antibody IP in SH-SY5Y cell lysate. Lane 3: Mouse IgG instead of Parkin Recombinant Rabbit Monoclonal Antibody in SH-SY5Y cell lysate. Blocking/Dilution buffer: 5% NFDM/TBST. Exposure time: 2 minutes.
IF-P
Application: Immunofluorescence (IF-tissue). Species: Mouse. Tissue: Hippocampus. Sample: Paraffin-embedded section. Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST. Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: Parkin Recombinant Rabbit Monoclonal Antibody, 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.
IF-P
Application: Immunofluorescence (IF-tissue). Species: Rat. Tissue: Brain. Sample: Paraffin-embedded section. Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃. Wash buffer: 1× TBST Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. Primary antibody: Parkin Recombinant Rabbit Monoclonal Antibody, 1/200, overnight at 4℃. Secondary antibody: Goat Anti-Rabbit IgG (488), 1.5 hours at room temperature.
IF-F
Application: IHC-Fr. Species: Mouse. Site: Cerebral cortex. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.
IF-F
Application: IHC-Fr. Species: Rat. Site: Cerebral cortex. Sample: Frozen section. Antibody concentration: 1/500. Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.| Application Notes | WB:1:2000-1:10000 IHC:1:1000 ICC/IF:1:50-1:100 IF-P:1:200 IF-F:1:500 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage Buffer | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
