Antibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [1]
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Validation data
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- Product number
- A-110 - Provider product page
- Provider
- R&D Systems
- Product name
- Human Phospho-Ubiquitin (S65) Antibody
- Antibody type
- Polyclonal
- Description
- Antigen Affinity-purified. This antibody detects mono- and poly-Ubiquitin chains that are phosphorylated on Serine 65. It has no cross-reactivity with non-phosphorylated Ubiquitin, Ubiquitin phosphorylated at Serine 57 or Tyrosine 59, or phosphorylated Parkin. Detects only recombinant, phosporylated Ubiquitin. Not recommended on natural lysates (see western blot results below).
- Reactivity
- Human
- Host
- Rabbit
- Conjugate
- Unconjugated
- Antigen sequence
P0CG47
- Isotype
- IgG
- Vial size
- 50 ug
- Storage
- 6 months from date of receipt, -20 °C as supplied. 3 months, -20 °C under sterile conditions after opening.
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Supportive validation
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- R&D Systems (provider)
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- Experimental details
- Detection 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 either alpha-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.