Human Phospho-Ubiquitin (S65) Antibody Summary
Applications
Please Note: Optimal dilutions should be determined by each laboratory for each application.General Protocolsare available in the Technical Information section on our website.
Data Examples
View LargerDetection of Phospho-Ubiquitin (S65) by Western blot.Tetraubiquitin chains of each indicated linkage type were incubated for 0-4 hours in reactions containing recombinant PINK1 kinase (Cat# AP-180) and ATP. At indicated times a portion of each reaction was removed and terminated with SDS-PAGE sample buffer. SDS-PAGE gels (10-20%) were used to resolve approximately 150 ng of Ubiquitin tetramer from each reaction. Western Blots were developed using eitheralpha -phospho-Ubiquitin, pS65 (upper panels) or anti-Ubiquitin (Cat# MAB701, lower panel). Primary antibodies were used at 1 μg/ml in PBST + 0.5% BSA, while HRP-labeled secondary antibodies ( alpha -rabbit for A-110,alpha -mouse for MAB701) were used at a 1:10,000 dilution in PBST + 0.5% BSA. Further details are available upon request.
View LargerDetection of Phospho-Ubiquitin (S65) by Western blot.Tetraubiquitin chains of each indicated linkage type were incubated for 0-4 hours in reactions containing recombinant PINK1 kinase (Cat# AP-180) and ATP. At indicated times a portion of each reaction was removed and terminated with SDS-PAGE sample buffer. SDS-PAGE gels (10-20%) were used to resolve approximately 150 ng of Ubiquitin tetramer from each reaction. Western Blots were developed using eitheralpha -phospho-Ubiquitin, pS65 (upper panels) or anti-Ubiquitin (Cat# MAB701, lower panel). Primary antibodies were used at 1 μg/ml in PBST + 0.5% BSA, while HRP-labeled secondary antibodies ( alpha -rabbit for A-110,alpha -mouse for MAB701) were used at a 1:10,000 dilution in PBST + 0.5% BSA. Further details are available upon request.
Reconstitution Calculator
Preparation and Storage
- 6 months from date of receipt, -20 °C as supplied.
- 3 months, -20 °C under sterile conditions after opening.
Background: Ubiquitin
Serine/Threonine kinase PINK1 (PTEN-induced putative kinase protein 1) plays a critical role in preventing mitochondrial dysfunction during cellular stress. PINK is translated in the cytosol, then translocated to the outer mitochondrial membrane where it is rapidly cleaved and degraded as a part of normal mitochondrial function. In damaged (depolarized) mitochondria PINK becomes stabilized and accumulates, resulting in the subsequent phosphorylation of numerous proteins on the mitochondrial surface including Mfn2. Ultimately PARK2 (E3 Ubiquitin Ligase Parkin) is recruited to the damaged mitochondria where it is activated by PINK-mediated phosphorylation of PARK2 at serine 65, and PARK2 interaction with phosphorylated Ubiquitin (also phosphorylated by PINK on serine 65). This signaling cascade is critical for clearing the damaged mitochondria via selective autophagy (mitophagy) by mediating activation and translocation of PARK2.
Product Datasheets
FAQs
No product specific FAQs exist for this product, however you may
View all Antibody FAQsIsotype Controls
Normal Rabbit IgG Control
Normal RabbitIgG Control
Secondary Antibodies
Donkey Anti-Rabbit IgG NorthernLights™ NL637-conjugated Antibody
Rabbit IgG Antibody
Donkey Anti-Rabbit IgG NorthernLights™ NL493-conjugated Antibody
Rabbit IgG PE-conjugated Antibody
Rabbit IgG APC-conjugated Antibody
Donkey Anti-Rabbit IgG NorthernLights™ NL557-conjugated Antibody
Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody
Rabbit IgG Fluorescein-conjugated Antibody
Donkey Anti-Rabbit IgG (H+L) Antibody
Rabbit IgG Biotinylated Antibody
Rabbit IgG HRP-conjugated Antibody
Rabbit IgG VisUCyte HRP Polymer Antibody
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