Western Blot (WB)
(Figure 1. Western blot analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).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 30ug of sample under reducing conditions.Lane 1: human U87 whole cell lysatesLane 2: human SH-SY5Y whole cell lysatesLane 3: rat brain tissue lysatesLane 4: mouse brain tissue 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 mouse anti- PGP9. 5 antigen affinity purified monoclonal antibody (Catalog # MBS1753987) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PGP9. 5 at approximately 27KD. The expected band size for PGP9. 5 is at 27KD. )
Mouse PGP9. 5 Monoclonal Antibody | anti-UCHL1 antibody
Anti-PGP9. 5 Antibody (monoclonal, 3E4)
IHC-P: 2-5ug/ml|Human, Rat|
FC/FACS/FCM: 1-3ug/1x106 cells|Human|
Western Blot (WB)
(Figure 1. Western blot analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).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 30ug of sample under reducing conditions.Lane 1: human U87 whole cell lysatesLane 2: human SH-SY5Y whole cell lysatesLane 3: rat brain tissue lysatesLane 4: mouse brain tissue 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 mouse anti- PGP9. 5 antigen affinity purified monoclonal antibody (Catalog # MBS1753987) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PGP9. 5 at approximately 27KD. The expected band size for PGP9. 5 is at 27KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).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 30ug of sample under reducing conditions.Lane 1: human U87 whole cell lysatesLane 2: human SH-SY5Y whole cell lysatesLane 3: rat brain tissue lysatesLane 4: mouse brain tissue 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 mouse anti- PGP9. 5 antigen affinity purified monoclonal antibody (Catalog # MBS1753987) 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # MBS176445) with Tanon 5200 system. A specific band was detected for PGP9. 5 at approximately 27KD. The expected band size for PGP9. 5 is at 27KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).PGP9. 5 was detected in paraffin-embedded section of human renal clear cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9. 5 Antibody (MBS1753987) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).PGP9. 5 was detected in paraffin-embedded section of human renal clear cell carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9. 5 Antibody (MBS1753987) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).PGP9. 5 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9. 5 Antibody (MBS1753987) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of PGP9. 5 using anti-PGP9. 5 antibody (MBS1753987).PGP9. 5 was detected in paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8. 0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-PGP9. 5 Antibody (MBS1753987) overnight at 4 degree C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176438) with DAB as the chromogen. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of 293T cells using anti-PGP9. 5 antibody (MBS1753987).Overlay histogram showing 293T cells stained with MBS1753987 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PGP9. 5 Antibody (MBS1753987, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Flow Cytometry (FC/FACS)
(Figure 4. Flow Cytometry analysis of 293T cells using anti-PGP9. 5 antibody (MBS1753987).Overlay histogram showing 293T cells stained with MBS1753987 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-PGP9. 5 Antibody (MBS1753987, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
2. Das C, Hoang QQ, Kreinbring CA, Luchansky SJ, Meray RK, Ray SS, Lansbury PT, Ringe D, Petsko GA (Mar 2006). "Structural basis for conformational plasticity of the Parkinson's disease-associated ubiquitin hydrolase UCH-L1". Proceedings of the National Academy of Sciences of the United States of America 103 (12): 4675-80.
3. Doran JF, Jackson P, Kynoch PA, Thompson RJ (Jun 1983). "Isolation of PGP 9. 5, a new human neurone-specific protein detected by high-resolution two-dimensional electrophoresis". Journal of Neurochemistry 40 (6): 1542-7.
NCBI and Uniprot Product Information
NCBI Description
The protein encoded by this gene belongs to the peptidase C12 family. This enzyme is a thiol protease that hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin. This gene is specifically expressed in the neurons and in cells of the diffuse neuroendocrine system. Mutations in this gene may be associated with Parkinson disease.[provided by RefSeq, Sep 2009]
Uniprot Description
UCHL1: Ubiquitin-protein hydrolase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. This enzyme is a thiol protease that recognizes and hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin. Also binds to free monoubiquitin and may prevent its degradation in lysosomes. The homodimer may have ATP-independent ubiquitin ligase activity. Monomer. Homodimer. Interacts with SNCA. Interacts with COPS5. Found in neuronal cell bodies and processes throughout the neocortex. Expressed in neurons and cells of the diffuse neuroendocrine system and their tumors. Weakly expressed in ovary. Down-regulated in brains from Parkinson disease and Alzheimer disease patients. Belongs to the peptidase C12 family.
Protein type: Cell development/differentiation; EC 3.4.19.12; Ligase; Protease; Ubiquitin conjugating system
Chromosomal Location of Human Ortholog: 4p14
Cellular Component: nucleoplasm; endoplasmic reticulum membrane; cell soma; axon; cytoplasm; plasma membrane; cytosol
Molecular Function: protein binding; omega peptidase activity; ubiquitin binding; cysteine-type endopeptidase activity; ubiquitin-specific protease activity; alpha-2A adrenergic receptor binding; ligase activity
Biological Process: proteasomal ubiquitin-dependent protein catabolic process; cell proliferation; negative regulation of MAP kinase activity; protein deubiquitination; axon transport of mitochondrion; axon target recognition; eating behavior; neuromuscular process; sensory perception of pain; muscle fiber development; adult walking behavior
Disease: Neurodegeneration With Optic Atrophy, Childhood-onset; Parkinson Disease 5, Autosomal Dominant
Research Articles on UCHL1
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Product Notes
The UCHL1 uchl1 (Catalog #AAA1753987) is an Antibody produced from Mouse and is intended for research purposes only. The product is available for immediate purchase. The Anti-PGP9. 5 Antibody (monoclonal, 3E4) reacts with Human, Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's PGP9. 5 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM). WB: 0.25-0.5ug/ml|Human, Mouse, Rat| IHC-P: 2-5ug/ml|Human, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Human|. Researchers should empirically determine the suitability of the UCHL1 uchl1 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 "PGP9. 5, Monoclonal 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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