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PRODUCT CODE: ET1702-60

Parkin Recombinant Rabbit Monoclonal Antibody [JF82-09] (ET1702-60)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

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Western blot analysis of Parkin on 293T cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-60, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Parkin on 293T cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-60, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • 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).
  • ICC staining of Parkin in SH-SY5Y 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).
  • 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).
  • 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.
  • 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.
  • Flow cytometric analysis of Parkin was done on SH-SY5Y cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-60, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Parkin on 293T cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-60, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • IP

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Parkin Recombinant Rabbit Monoclonal Antibody [JF82-09] (ET1702-60)

Immunogen

Synthetic peptide within n-terminal human parkin.

Host

Rabbit

Positive Control

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

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JF82-09

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

52 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • 1:50

TARGET

UNIPROT #

O60260

PROTEIN NAME

Parkin

SYNONYMS

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

SEQUENCE SIMILARITIES

Belongs to the RBR family. Parkin subfamily.

TISSUE SPECIFICITY

Highly expressed in the brain including the substantia nigra. Expressed in heart, testis and skeletal muscle. Expression is down-regulated or absent in tumor biopsies, and absent in the brain of PARK2 patients. Overexpression protects dopamine neurons from kainate-mediated apoptosis. Found in serum (at protein level).

POST-TRANSLATIONAL MODIFICATION

Auto-ubiquitinates in an E2-dependent manner leading to its own degradation. Also polyubiquitinated by RNF41 for proteasomal degradation.; S-nitrosylated. The inhibition of PRKN ubiquitin E3 ligase activity by S-nitrosylation could contribute to the degenerative process in PD by impairing the ubiquitination of PRKN substrates.; Phosphorylation at Ser-65 by PINK1 contributes to activate PRKN activity. It is however not sufficient and requires binding to phosphorylated ubiquitin as well.

SUBCELLULAR LOCATION

Mitochondrion, cytosol, Nucleus, Endoplasmic reticulum.

FUNCTION

Parkin is a zinc-finger protein that is related to ubiquitin at the amino terminus. The wild type Parkin gene, which maps to human chromosome 6q25.2-27, encodes a 465 amino acid full-length protein that is expressed as multiple isoforms. Mutations in the Parkin gene are responsible for autosomal recessive juvenile Parkinson's disease and commonly involve deletions of exons 3-5. In humans, Parkin is expressed in a subset of cells of the basal ganglia, midbrain, cerebellum and cerebral cortex, and is subject to alternative splicing in different tissues. Parkin expression is also high in the brainstem of mice, with the majority of immunopositive cells being neurons. The Parkin gene has been identified in a diverse group of organisms including mammals, birds, frog and fruit flies, suggesting that analogous functional roles of the Parkin protein may have been highly conserved during the course of evolution.

CITATIONS

  • Lei, Zhong et al.

    PARK2 inhibits osteosarcoma cell growth through the JAK2/STAT3/VEGF signaling pathway. | Cell Death & Disease [2018]