Anti-Parkin antibody [PRK8] ab77924 is a mouse monoclonal antibody that is used in Parkin western blotting. Suitable for human, mouse and rat samples.
- Antibody clone PRK8 is the most widely used clone for Parkin on the market and is cited in >520 publications
- Specificity confirmed with PRKN knockout cell line validation
IgG2b
Mouse
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Liquid
Monoclonal
Flow Cyt | WB | |
---|---|---|
Human | Not recommended | Tested |
Mouse | Not recommended | Tested |
Rat | Not recommended | Tested |
Drosophila melanogaster | Not recommended | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human, Drosophila melanogaster | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species Rat | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species Human | Dilution info 1/1000 - 1/2000 | Notes Abcam recommends using 1-3% Milk as the blocking agent. Higher percentage blocking solutions may not give optimal results. |
Species | Dilution info | Notes |
---|---|---|
Species Drosophila melanogaster | Dilution info - | Notes - |
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Functions within a multiprotein E3 ubiquitin ligase complex, catalyzing the covalent attachment of ubiquitin moieties onto substrate proteins (PubMed:10888878, PubMed:10973942, PubMed:11431533, PubMed:12150907, PubMed:12628165, PubMed:15105460, PubMed:16135753, PubMed:21376232, PubMed:21532592, PubMed:22396657, PubMed:23620051, PubMed:23754282, PubMed:24660806, PubMed:24751536, PubMed:29311685, PubMed:32047033). Substrates include SYT11 and VDAC1 (PubMed:29311685, PubMed:32047033). Other substrates are BCL2, CCNE1, GPR37, RHOT1/MIRO1, MFN1, MFN2, STUB1, SNCAIP, SEPTIN5, TOMM20, USP30, ZNF746, MIRO1 and AIMP2 (PubMed:10888878, PubMed:10973942, PubMed:11431533, PubMed:12150907, PubMed:12628165, PubMed:15105460, PubMed:16135753, PubMed:21376232, PubMed:21532592, PubMed:22396657, PubMed:23620051, PubMed:23754282, PubMed:24660806, PubMed:24751536). Mediates monoubiquitination as well as 'Lys-6', 'Lys-11', 'Lys-48'-linked and 'Lys-63'-linked polyubiquitination of substrates depending on the context (PubMed:19229105, PubMed:20889974, PubMed:25474007, PubMed:25621951, PubMed:32047033). 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 (PubMed:17846173, PubMed:19229105). Mediates 'Lys-63'-linked polyubiquitination of a 22 kDa O-linked glycosylated isoform of SNCAIP, possibly playing a role in Lewy-body formation (PubMed:11431533, PubMed:11590439, PubMed:15105460, PubMed:15728840, PubMed:19229105). Mediates monoubiquitination of BCL2, thereby acting as a positive regulator of autophagy (PubMed:20889974). Protects against mitochondrial dysfunction during cellular stress, by acting downstream of PINK1 to coordinate mitochondrial quality control mechanisms that remove and replace dysfunctional mitochondrial components (PubMed:11439185, PubMed:18957282, PubMed:19029340, PubMed:19966284, PubMed:21376232, PubMed:22082830, PubMed:22396657, PubMed:23620051, PubMed:23933751, PubMed:24660806, PubMed:24784582, PubMed:24896179, PubMed:25474007, PubMed:25527291, PubMed:32047033). Depending on the severity of mitochondrial damage and/or dysfunction, activity ranges from preventing apoptosis and stimulating mitochondrial biogenesis to regulating mitochondrial dynamics and eliminating severely damaged mitochondria via mitophagy (PubMed:11439185, PubMed:19029340, PubMed:19801972, PubMed:19966284, PubMed:21376232, PubMed:22082830, PubMed:22396657, PubMed:23620051, PubMed:23685073, PubMed:23933751, PubMed:24896179, PubMed:25527291, PubMed:32047033, PubMed:33499712). Activation and recruitment onto the outer membrane of damaged/dysfunctional mitochondria (OMM) requires PINK1-mediated phosphorylation of both PRKN and ubiquitin (PubMed:24660806, PubMed:24784582, PubMed:25474007, PubMed:25527291). After mitochondrial damage, functions with PINK1 to mediate the decision between mitophagy or preventing apoptosis by inducing either the poly- or monoubiquitination of VDAC1, respectively; polyubiquitination of VDAC1 promotes mitophagy, while monoubiquitination of VDAC1 decreases mitochondrial calcium influx which ultimately inhibits apoptosis (PubMed:27534820, PubMed:32047033). When cellular stress results in irreversible mitochondrial damage, promotes the autophagic degradation of dysfunctional depolarized mitochondria (mitophagy) by promoting the ubiquitination of mitochondrial proteins such as TOMM20, RHOT1/MIRO1, MFN1 and USP30 (PubMed:19029340, PubMed:19966284, PubMed:21753002, PubMed:22396657, PubMed:23620051, PubMed:23685073, PubMed:23933751, PubMed:24896179, PubMed:25527291). Preferentially assembles 'Lys-6'-, 'Lys-11'- and 'Lys-63'-linked polyubiquitin chains, leading to mitophagy (PubMed:25621951, PubMed:32047033). The PINK1-PRKN pathway also promotes fission of damaged mitochondria by PINK1-mediated phosphorylation which promotes the PRKN-dependent degradation of mitochondrial proteins involved in fission such as MFN2 (PubMed:23620051). This prevents the refusion of unhealthy mitochondria with the mitochondrial network or initiates mitochondrial fragmentation facilitating their later engulfment by autophagosomes (PubMed:23620051). Regulates motility of damaged mitochondria via the ubiquitination and subsequent degradation of MIRO1 and MIRO2; in motor neurons, this likely inhibits mitochondrial intracellular anterograde transport along the axons which probably increases the chance of the mitochondria undergoing mitophagy in the soma (PubMed:22396657). Involved in mitochondrial biogenesis via the 'Lys-48'-linked polyubiquitination of transcriptional repressor ZNF746/PARIS which leads to its subsequent proteasomal degradation and allows activation of the transcription factor PPARGC1A (PubMed:21376232). Limits the production of reactive oxygen species (ROS) (PubMed:18541373). Regulates cyclin-E during neuronal apoptosis (PubMed:12628165). In collaboration with CHPF isoform 2, may enhance cell viability and protect cells from oxidative stress (PubMed:22082830). Independently of its ubiquitin ligase activity, protects from apoptosis by the transcriptional repression of p53/TP53 (PubMed:19801972). May protect neurons against alpha synuclein toxicity, proteasomal dysfunction, GPR37 accumulation, and kainate-induced excitotoxicity (PubMed:11439185). May play a role in controlling neurotransmitter trafficking at the presynaptic terminal and in calcium-dependent exocytosis. May represent a tumor suppressor gene (PubMed:12719539).
PARK2, PRKN, E3 ubiquitin-protein ligase parkin, Parkin, Parkin RBR E3 ubiquitin-protein ligase, Parkinson juvenile disease protein 2, Parkinson disease protein 2
Anti-Parkin antibody [PRK8] ab77924 is a mouse monoclonal antibody that is used in Parkin western blotting. Suitable for human, mouse and rat samples.
- Antibody clone PRK8 is the most widely used clone for Parkin on the market and is cited in >520 publications
- Specificity confirmed with PRKN knockout cell line validation
IgG2b
Mouse
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Liquid
Monoclonal
PRK8
Affinity purification Protein G
The epitope is the second ring domain (aa 399-465).
kappa
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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This supplementary information is collated from multiple sources and compiled automatically.
The Parkin protein also known as PRK8 or Park2 is an E3 ubiquitin ligase with a molecular weight of approximately 52 kDa. This protein plays a critical role in tagging damaged proteins for degradation maintaining cellular health. Parkin is expressed in various tissues with significant levels in dopaminergic neurons in the brain. It is encoded by the PARK2 gene and has been linked to the regulation of mitochondrial quality and autophagy processes contributing to cellular homeostasis.
Parkin is essential for the regulation of mitochondria through its involvement in the mitochondrial quality control system. It functions as part of a complex with other proteins that respond to mitochondrial damage by tagging them with ubiquitin molecules. This mechanism allows for the removal of defective mitochondria via mitophagy critical for preventing the accumulation of damaged cellular components.
Parkin interacts with pathways involved in the cellular stress response particularly the PINK1 (PTEN Induced Kinase 1) pathway. PINK1 phosphorylates Parkin activating it to label damaged mitochondria. Another critical pathway involves proteasomal degradation where Parkin collaborates with Ubiquitin to manage protein turnover. These pathways highlight its relationships with other cellular stress-regulating proteins enhancing our understanding of its roles in maintaining cellular integrity.
Mutations in the gene coding for Parkin are linked to Parkinson's disease (PD) and some forms of juvenile autosomal recessive parkinsonism. The Parkin protein's dysfunctional activity leads to impaired mitochondrial management and protein aggregation in neurons contributing significantly to neurodegenerative disease. In conditions such as PD Parkin interacts with other proteins such as PINK1 reinforcing its role in mitochondrial protection and indicating the protein's importance in disease progression and potential therapeutic targeting.
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This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
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Terms & Conditions.
Lanes 1 - 4: Merged signal (red and green). Green - ab77924 observed at 49 kDa. Red - loading control Anti-GAPDH antibody [EPR16891] - 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 Human PARK2 (Parkin) knockout SH-SY5Y cell line ab280042 (PRKN knockout cell lysate Human PARK2 (Parkin) knockout SH-SY5Y 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 Anti-GAPDH antibody [EPR16891] - Loading Control 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 (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/mL
Lane 1: Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2: PRKN knockout SH-SY5Y cell lysate at 20 µg
Lane 3: Human Brain tissue lysate at 20 µg
Lane 4: HUVEC cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 52 kDa
Observed band size: 49 kDa
Blocking buffer: 3% milk.
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 5 µg/mL
Lane 1: Human brain tissue lysate at 20 µg
Lane 2: Mouse brain tissue lysate at 20 µg
Lane 3: Rat brain tissue lysate at 20 µg
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: 8min
All lanes blocked with 3% milk.
All lanes: Western blot - Anti-Parkin antibody [PRK8] (ab77924) at 1/2000 dilution
Lane 1: SH-SY5Y (Human neuroblastoma cell line) Whole Cell Lysate at 20 µg
Lane 2: Brain (Rat) Tissue Lysate at 20 µg
Lane 3: Brain (Mouse) Tissue Lysate at 20 µg
Lane 4: Brain (Human) Tissue Lysate at 20 µg
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
Exposure time: 20min
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