A-110

antibody from R&D Systems

Targeting: RPS27A S27A, Uba80, UBCEP80

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

A-110

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.

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

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.