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Anti-Phospho-Ubiquitin (Ser65) Rabbit Antibody [C22F3]

Catalog No.: F1476

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IP1:100

Application
WB, IP

Reactivity
Human

Source
Rabbit

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Positive Control	PC-3 (carbonyl cyanide 3-chlorophenylhydrazone, 30 μM, 6 h)
Negative Control	PC-3

Datasheet & SDS

Biological Description

Specificity
Phospho-Ubiquitin (Ser65) Rabbit mAb recognizes endogenous levels of Ubiquitin protein only when phosphorylated at Ser65.

Clone
C22F3

Synonym(s)
Ubiquitin-ribosomal protein eL40 fusion protein, CEP52, Ubiquitin A-52 residue ribosomal protein fusion product 1, UBA52, UBCEP2, Polyubiquitin-C, UBC, Polyubiquitin-B, UBB, Ubiquitin-ribosomal protein eS31 fusion protein, Ubiquitin carboxyl extension protein 80, RPS27A, UBA80, UBCEP1

Background
Ubiquitin is a highly conserved polypeptide that plays a central role in the ubiquitin-proteasome system, a key mechanism for regulating protein degradation within cells. Through a process known as ubiquitination, ubiquitin molecules are covalently attached to specific cellular proteins, marking them for degradation by the 26S proteasome. The ubiquitination cascade involves three main enzymatic steps: First, ubiquitin is activated by the E1 enzyme, forming a thiolester bond. This activated ubiquitin is then transferred to the E2 ubiquitin-conjugating enzyme. Finally, the E3 ubiquitin ligase facilitates the transfer of ubiquitin from E2 to the target protein by attaching it to the epsilon-amino group of a lysine residue on the substrate. The ubiquitin-proteasome pathway is critical for a wide array of physiological processes, including cell cycle regulation, differentiation, response to cellular stress, and apoptosis. Dysregulation of this pathway has been linked to various diseases. One regulatory mechanism involves the phosphorylation of ubiquitin at serine 65 by the kinase PINK1, which activates the E3 ligase Parkin—a key step in mitochondrial quality control. IκB, p53, cdc25A, and Bcl-2, are known substrates of the ubiquitin-proteasome system, highlighting its essential role in maintaining cellular homeostasis and controlling vital functions such as immune responses, DNA repair, and programmed cell death.

Background

Ubiquitin is a highly conserved polypeptide that plays a central role in the ubiquitin-proteasome system, a key mechanism for regulating protein degradation within cells. Through a process known as ubiquitination, ubiquitin molecules are covalently attached to specific cellular proteins, marking them for degradation by the 26S proteasome. The ubiquitination cascade involves three main enzymatic steps: First, ubiquitin is activated by the E1 enzyme, forming a thiolester bond. This activated ubiquitin is then transferred to the E2 ubiquitin-conjugating enzyme. Finally, the E3 ubiquitin ligase facilitates the transfer of ubiquitin from E2 to the target protein by attaching it to the epsilon-amino group of a lysine residue on the substrate. The ubiquitin-proteasome pathway is critical for a wide array of physiological processes, including cell cycle regulation, differentiation, response to cellular stress, and apoptosis. Dysregulation of this pathway has been linked to various diseases. One regulatory mechanism involves the phosphorylation of ubiquitin at serine 65 by the kinase PINK1, which activates the E3 ligase Parkin—a key step in mitochondrial quality control. IκB, p53, cdc25A, and Bcl-2, are known substrates of the ubiquitin-proteasome system, highlighting its essential role in maintaining cellular homeostasis and controlling vital functions such as immune responses, DNA repair, and programmed cell death.

References
https://pubmed.ncbi.nlm.nih.gov/10939967/ https://pubmed.ncbi.nlm.nih.gov/24751536/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

Tel: +1-832-582-8158 Ext:3
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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%