Anti-Phospho-cdc25C (Thr48) Rabbit Antibody [G24N3]

Catalog No.: F1949

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 IP1:100 IF1:300

Application
WB, IP, IF

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

Predicted MW
75 kDa

Positive Control	HT-29 (synchronized in mitosis (thymidine block, nocodazole release, mitotic shake-off)
Negative Control

Datasheet & SDS

Biological Description

Specificity
Phospho-cdc25C (Thr48) Rabbit mAb recognizes endogenous levels of cdc25C only when phosphorylated at Thr48.

Clone
G24N3

Synonym(s)
M-phase inducer phosphatase 3, Dual specificity phosphatase Cdc25C, CDC25C

Background
Phospho-CDC25C (Thr48) refers to the phosphorylation of the CDC25C protein at the threonine 48 residue, a key post-translational modification within its N-terminal regulatory domain. CDC25C is a dual-specificity phosphatase that plays a critical role in driving the cell cycle progression from G2 phase into mitosis by activating the cyclin B1/CDK1 complex. Phosphorylation at Thr48 creates a specific recognition site for the prolyl isomerase Pin1, which binds to this phosphorylated motif and catalyzes a conformational change in CDC25C. This conformational shift promotes the dephosphorylation of CDC25C by phosphoprotein phosphatase 2A (PP2A), leading to its activation and nuclear translocation, thereby facilitating the activation of the cyclin B1/CDK1 complex and mitotic entry. Phosphorylation at Thr48 and its subsequent isomerization by Pin1 are essential for proper G2/M transition and mitotic progression. Dysregulation of this pathway, including abnormal phosphorylation or altered Pin1 activity, has been linked to uncontrolled cell proliferation and tumorigenesis.

Background

Phospho-CDC25C (Thr48) refers to the phosphorylation of the CDC25C protein at the threonine 48 residue, a key post-translational modification within its N-terminal regulatory domain. CDC25C is a dual-specificity phosphatase that plays a critical role in driving the cell cycle progression from G2 phase into mitosis by activating the cyclin B1/CDK1 complex. Phosphorylation at Thr48 creates a specific recognition site for the prolyl isomerase Pin1, which binds to this phosphorylated motif and catalyzes a conformational change in CDC25C. This conformational shift promotes the dephosphorylation of CDC25C by phosphoprotein phosphatase 2A (PP2A), leading to its activation and nuclear translocation, thereby facilitating the activation of the cyclin B1/CDK1 complex and mitotic entry. Phosphorylation at Thr48 and its subsequent isomerization by Pin1 are essential for proper G2/M transition and mitotic progression. Dysregulation of this pathway, including abnormal phosphorylation or altered Pin1 activity, has been linked to uncontrolled cell proliferation and tumorigenesis.

References
https://pubmed.ncbi.nlm.nih.gov/27038604/ https://pubmed.ncbi.nlm.nih.gov/32518522/

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

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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%