Anti-Parkin antibody [PRK8] (ab77924)
![Western blot - Anti-Parkin antibody [PRK8] (ab77924)](https://www.abcam.com/ps/products/77/ab77924/Images/ab77924-469680-anti-parkin-antibody-prk8-western-blot-wildtype-shsy5y-prkn-knockout-shsy5y-human-brain-huvec.jpg "Western blot - Anti-Parkin antibody [PRK8] (ab77924)")
Key features and details
- Mouse monoclonal [PRK8] to Parkin
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
- Isotype: IgG2b
Overview
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Product name
Anti-Parkin antibody [PRK8]
See all Parkin primary antibodies -
Description
Mouse monoclonal [PRK8] to Parkin -
Host species
Mouse -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Drosophila melanogaster
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Immunogen
Recombinant full length protein corresponding to Human Parkin.
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Epitope
The epitope is the second ring domain (aa 399-465). -
Positive control
- WB: SH-SY5Y and HUVEC whole cell lysate. Human, mouse and rat brain tissue lysates.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading... -
Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
PRK8 -
Isotype
IgG2b -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab77924 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (7) |
1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa).
Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
| Notes |
|---|
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WB
1/2000. Detects a band of approximately 55 kDa (predicted molecular weight: 52 kDa). Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Application notes
Is unsuitable for Flow Cyt.
Target
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Function
Functions within a multiprotein E3 ubiquitin ligase complex, catalyzing the covalent attachment of ubiquitin moieties onto substrate proteins, such as BCL2, SYT11, CCNE1, GPR37, STUB1, a 22 kDa O-linked glycosylated isoform of SNCAIP, SEPT5, ZNF746 and AIMP2. Mediates monoubiquitination as well as 'Lys-48'-linked and 'Lys-63'-linked polyubiquitination of substrates depending on the context. Participates in the removal and/or detoxification of abnormally folded or damaged protein by mediating 'Lys-63'-linked polyubiquitination of misfolded proteins such as PARK7: 'Lys-63'-linked polyubiquitinated misfolded proteins are then recognized by HDAC6, leading to their recruitment to aggresomes, followed by degradation. Mediates 'Lys-63'-linked polyubiquitination of SNCAIP, possibly playing a role in Lewy-body formation. Mediates monoubiquitination of BCL2, thereby acting as a positive regulator of autophagy. Promotes the autophagic degradation of dysfunctional depolarized mitochondria. Mediates 'Lys-48'-linked polyubiquitination of ZNF746, followed by degradation of ZNF746 by the proteasome; possibly playing a role in role in regulation of neuron death. Limits the production of reactive oxygen species (ROS). Loss of this ubiquitin ligase activity appears to be the mechanism underlying pathogenesis of PARK2. May protect neurons against alpha synuclein toxicity, proteasomal dysfunction, GPR37 accumulation, and kainate-induced excitotoxicity. May play a role in controlling neurotransmitter trafficking at the presynaptic terminal and in calcium-dependent exocytosis. Regulates cyclin-E during neuronal apoptosis. May represent a tumor suppressor gene. -
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). -
Pathway
Protein modification; protein ubiquitination. -
Involvement in disease
Defects in PARK2 are a cause of Parkinson disease (PARK) [MIM:168600]. A complex neurodegenerative disorder characterized by bradykinesia, resting tremor, muscular rigidity and postural instability. Additional features are characteristic postural abnormalities, dysautonomia, dystonic cramps, and dementia. The pathology of Parkinson disease involves the loss of dopaminergic neurons in the substantia nigra and the presence of Lewy bodies (intraneuronal accumulations of aggregated proteins), in surviving neurons in various areas of the brain. The disease is progressive and usually manifests after the age of 50 years, although early-onset cases (before 50 years) are known. The majority of the cases are sporadic suggesting a multifactorial etiology based on environmental and genetic factors. However, some patients present with a positive family history for the disease. Familial forms of the disease usually begin at earlier ages and are associated with atypical clinical features.
Defects in PARK2 are the cause of Parkinson disease type 2 (PARK2) [MIM:600116]; also known as early-onset parkinsonism with diurnal fluctuation (EPDF) or autosomal recessive juvenile Parkinson disease (PDJ). A neurodegenerative disorder characterized by bradykinesia, rigidity, postural instability, tremor, and onset usually befor 40. It differs from classic Parkinson disease by early DOPA-induced dyskinesia, diurnal fluctuation of the symptoms, sleep benefit, dystonia and hyper-reflexia. Dementia is absent. Pathologically, patients show loss of dopaminergic neurons in the substantia nigra, similar to that seen in Parkinson disease; however, Lewy bodies (intraneuronal accumulations of aggregated proteins) are absent.
Note=Defects in PARK2 may be involved in the development and/or progression of ovarian cancer. -
Sequence similarities
Belongs to the RBR family. Parkin subfamily.
Contains 1 IBR-type zinc finger.
Contains 2 RING-type zinc fingers.
Contains 1 ubiquitin-like domain. -
Domain
The ubiquitin-like domain binds the PSMD4 subunit of 26S proteasomes. -
Post-translational
modificationsAuto-ubiquitinates in an E2-dependent manner leading to its own degradation. Also polyubiquitinated by RNF41 for proteasomal degradation.
S-nitrosylated. The inhibition of PARK2 ubiquitin E3 ligase activity by S-nitrosylation could contribute to the degenerative process in PD by impairing the ubiquitination of PARK2 substrates. -
Cellular localization
Cytoplasm > cytosol. Nucleus. Endoplasmic reticulum. Mitochondrion. Mainly localizes in the cytosol. Co-localizes with SYT11 in neutrites. Co-localizes with SNCAIP in brainstem Lewy bodies. Relocates to dysfunctional mitochondria that have lost the mitochondial membrane potential; recruitement to mitochondria is PINK1-dependent. - Information by UniProt
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Database links
- Entrez Gene: 5071 Human
- Entrez Gene: 50873 Mouse
- Entrez Gene: 56816 Rat
- Omim: 602544 Human
- SwissProt: O60260 Human
- SwissProt: Q9WVS6 Mouse
- SwissProt: Q9JK66 Rat
- Unigene: 132954 Human
see all -
Alternative names
- AR JP antibody
- E3 ubiquitin ligase antibody
- E3 ubiquitin protein ligase parkin antibody
see all
Images
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Western blot - Anti-Parkin antibody [PRK8] (ab77924)All lanes : Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/ml
Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : PRKN knockout SH-SY5Y cell lysate
Lane 3 : Human Brain tissue lysate
Lane 4 : HUVEC cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 49 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab77924 observed at 49 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab77924 was shown to react with Parkin in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PRKN knockout cell line ab280042 (PRKN knockout cell lysate ab280101). Wild-type SH-SY5Y and PRKN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab77924 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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![Western blot - Anti-Parkin antibody [PRK8] (ab77924)](https://www.abcam.com/ps/products/77/ab77924/Images/ab77924-465374-anti-parkin-antibody-prk8-western-blot-human-mouse-rat.jpg)
Western blot - Anti-Parkin antibody [PRK8] (ab77924)All lanes : Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/ml
Lane 1 : Human brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 8 minutesBlocking buffer: 3% milk.
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![Western blot - Anti-Parkin antibody [PRK8] (ab77924)](https://www.abcam.com/ps/products/77/ab77924/Images/ab77924-169148-ab77924GR1239510-WB.jpg)
Western blot - Anti-Parkin antibody [PRK8] (ab77924)All lanes : Anti-Parkin antibody [PRK8] (ab77924) at 1/2000 dilution
Lane 1 : SH-SY5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 2 : Brain (Rat) Tissue Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 : Brain (Human) Tissue Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutesAll lanes blocked with 3% milk.
Datasheets and documents
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SDS download
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Datasheet download
References (101)
ab77924 has been referenced in 101 publications.
- Choi GE et al. BNIP3L/NIX-mediated mitophagy protects against glucocorticoid-induced synapse defects. Nat Commun 12:487 (2021). PubMed: 33473105
- Zhou B et al. Mitochondrial quality control protects photoreceptors against oxidative stress in the H2O2-induced models of retinal degeneration diseases. Cell Death Dis 12:413 (2021). PubMed: 33879768
- Liu D et al. c-MYC-induced long noncoding RNA MEG3 aggravates kidney ischemia-reperfusion injury through activating mitophagy by upregulation of RTKN to trigger the Wnt/ß-catenin pathway. Cell Death Dis 12:191 (2021). PubMed: 33602903
- Roca-Agujetas V et al. Cholesterol alters mitophagy by impairing optineurin recruitment and lysosomal clearance in Alzheimer's disease. Mol Neurodegener 16:15 (2021). PubMed: 33685483
- Castro-Gonzalez S et al. HIV-1 Nef counteracts autophagy restriction by enhancing the association between BECN1 and its inhibitor BCL2 in a PRKN-dependent manner. Autophagy 17:553-577 (2021). PubMed: 32097085