Western Blot (WB)
(Figure 1. Western blot analysis of PINK1 using anti-PINK1 antibody (MBS1753293).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human HELA whole cell lysatesLane 3: human HEPG2 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-PINK1antigen affinity purified polyclonal antibody (Catalog # MBS1753293) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for PINK1 at approximately 64KD. The expected band size for PINK1 is at 64KD. )
Rabbit anti-Human PINK1 Polyclonal Antibody | anti-PINK1 antibody
Anti-PINK1 Antibody
ICC/IF: 5ug/ml|Human|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Direct ELISA: 0.1-0.5ug/ml|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of PINK1 using anti-PINK1 antibody (MBS1753293).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human HELA whole cell lysatesLane 3: human HEPG2 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-PINK1antigen affinity purified polyclonal antibody (Catalog # MBS1753293) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for PINK1 at approximately 64KD. The expected band size for PINK1 is at 64KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PINK1 using anti-PINK1 antibody (MBS1753293).Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.Lane 1: human A431 whole cell lysatesLane 2: human HELA whole cell lysatesLane 3: human HEPG2 whole cell lysates.After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1. 5 hour at RT. The membrane was incubated with rabbit anti-PINK1antigen affinity purified polyclonal antibody (Catalog # MBS1753293) at 0. 5 μg/mL overnight at 4 degree C, then washed with TBS-0. 1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176460) with Tanon 5200 system. A specific band was detected for PINK1 at approximately 64KD. The expected band size for PINK1 is at 64KD. )
Immunofluorescence (IF)
(Figure 2. IF analysis of PINK1 using anti- PINK1 antibody (MBS1753293).PINK1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-PINK1 Antibody (MBS1753293) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Immunofluorescence (IF)
(Figure 2. IF analysis of PINK1 using anti- PINK1 antibody (MBS1753293).PINK1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (MBS176582) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-PINK1 Antibody (MBS1753293) overnight at 4 degree C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37 degree C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of 293T cells using anti-PINK1 antibody (MBS1753293).Overlay histogram showing 293T cells stained with MBS1753293 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PINK1 Antibody (MBS1753293, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 3. Flow Cytometry analysis of 293T cells using anti-PINK1 antibody (MBS1753293).Overlay histogram showing 293T cells stained with MBS1753293 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PINK1 Antibody (MBS1753293, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Beilina, A., Van Der Brug, M., Ahmad, R., Kesavapany, S., Miller, D. W., Petsko, G. A., Cookson, M. R. Mutations in PTEN-induced putative kinase 1 associated with recessive parkinsonism have differential effects on protein stability. Proc. Nat. Acad. Sci. 102: 5703-5708, 2005.
3. Bingol, B., Tea, J. S., Phu, L., Reichelt, M., Bakalarski, C. E., Song, Q., Foreman, O., Kirkpatrick, D. S., Sheng, M. The mitochondrial deubiquitinase USP30 opposes parkin-mediated mitophagy. Nature 510: 370-375, 2014.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a serine/threonine protein kinase that localizes to mitochondria. It is thought to protect cells from stress-induced mitochondrial dysfunction. Mutations in this gene cause one form of autosomal recessive early-onset Parkinson disease. [provided by RefSeq, Jul 2008]
Uniprot Description
PINK1: Protects against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondrial proteins. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). It is necessary for PARK2 recruitment to dysfunctional mitochondria to initiate their degradation. Interats with PARK2. Highly expressed in heart, skeletal muscle and testis, and at lower levels in brain, placenta, liver, kidney, pancreas, prostate, ovary and small intestine. Present in the embryonic testis from an early stage of development. Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. 2 isoforms of the human protein are produced by alternative splicing.
Protein type: Kinase, protein; Membrane protein, integral; Protein kinase, Ser/Thr (non-receptor); Mitochondrial; EC 2.7.11.1; Protein kinase, Other; Other group; NKF2 family
Chromosomal Location of Human Ortholog: 1p36
Cellular Component: mitochondrion; mitochondrial intermembrane space; cytosol; mitochondrial outer membrane; cytoskeleton; membrane; axon; perinuclear region of cytoplasm; cytoplasm; mitochondrial inner membrane; TORC2 complex; chromatin; nucleus; integral to mitochondrial outer membrane
Molecular Function: protein kinase B binding; protein serine/threonine kinase activity; protein binding; protease activator activity; protease binding; ubiquitin protein ligase binding; magnesium ion binding; kinase activity; ATP binding
Biological Process: ubiquitin-dependent protein catabolic process; negative regulation of JNK cascade; positive regulation of dopamine secretion; positive regulation of translation; regulation of protein ubiquitination; protein ubiquitination; protein amino acid phosphorylation; positive regulation of protein amino acid dephosphorylation; negative regulation of macroautophagy; regulation of mitochondrial membrane potential; positive regulation of ubiquitin-protein ligase activity; response to stress; regulation of protein complex assembly; negative regulation of neuron apoptosis; regulation of hydrogen peroxide metabolic process; mitochondrion organization and biogenesis; positive regulation of I-kappaB kinase/NF-kappaB cascade; protein stabilization; activation of protein kinase B; positive regulation of synaptic transmission, dopaminergic; positive regulation of peptidyl-serine phosphorylation; positive regulation of protein kinase B signaling cascade; peptidyl-serine phosphorylation; mitochondrion degradation; positive regulation of transcription factor activity; positive regulation of protein amino acid phosphorylation; response to oxidative stress; phosphorylation
Disease: Parkinson Disease 6, Autosomal Recessive Early-onset
Research Articles on PINK1
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Product Notes
The PINK1 pink1 (Catalog #AAA1753293) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-PINK1 Antibody reacts with Human and may cross-react with other species as described in the data sheet. AAA Biotech's PINK1 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Human| ICC/IF: 5ug/ml|Human| FC/FACS/FCM: 1-3ug/1x106 cells|Human| Direct ELISA: 0.1-0.5ug/ml|Human|. Researchers should empirically determine the suitability of the PINK1 pink1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "PINK1, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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