Phospho-p130 (Ser672) Antibody [E21C6]

Application: Reactivity: Human

Usage Information

Dilution
WB1:5000 - 1:10000 IP1:20

Application
WB, IP

Reactivity
Human

Source
Rabbit Monoclonal Antibody

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 Observed MW
128 kDa 128 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight.

Positive Control	Jurkat cells
Negative Control	Jurkat cells (AP treated)

Datasheet & SDS

Biological Description

Specificity
Phospho-p130 (Ser672) Antibody [E21C6] detects endogenous levels of total p130 protein only when it is phosphorylated at Ser672.

Clone
E21C6

Synonym(s)
RB2; RBL2; Retinoblastoma-like protein 2; 130 kDa retinoblastoma-associated protein; Retinoblastoma-related protein 2; pRb2; p130; RBR-2

Background
Phospho-p130 (Ser672) refers to the retinoblastoma-related protein p130 (RBL2), a crucial tumor suppressor of the pocket protein family (including pRb and p107) that enforces G0/G1 cell cycle arrest by binding and repressing E2F transcription factors. p130 contains conserved N-terminal, spacer, and C-terminal regions, featuring a large A/B pocket domain (the cyclin-binding motif) and multiple phosphorylation sites targeted by cyclin-dependent kinases (CDKs). Among these, Ser672 in the C-terminus is a CDK4/6-specific phosphorylation site pivotal for proteasomal degradation. During the late G1/S phase, phosphorylation at Ser672 by CDK4/6 marks p130 for recognition by the F-box protein Skp2, a component of the SCF^Skp2 ubiquitin ligase complex. This enables p130 polyubiquitination and rapid proteasomal turnover, independently of CDK2 activity. As a result, p130-mediated repression of E2F-responsive genes is relieved, allowing S-phase entry, DNA replication, and cell proliferation, while terminating the growth-suppressive function of p130. In quiescent G0 cells, hypophosphorylated p130 forms stable complexes with E2F4/5-DP1 to silence proliferation-associated genes; phosphorylation at Ser672 dynamically lowers p130 levels as cells re-enter the cycle. Dysregulation of the CDK4/6-Skp2-p130 axis or p130 mutations can stabilize hyperphosphorylated forms and disrupt cell cycle checkpoints, promoting uncontrolled proliferation and cancer development.

Background

Phospho-p130 (Ser672) refers to the retinoblastoma-related protein p130 (RBL2), a crucial tumor suppressor of the pocket protein family (including pRb and p107) that enforces G0/G1 cell cycle arrest by binding and repressing E2F transcription factors. p130 contains conserved N-terminal, spacer, and C-terminal regions, featuring a large A/B pocket domain (the cyclin-binding motif) and multiple phosphorylation sites targeted by cyclin-dependent kinases (CDKs). Among these, Ser672 in the C-terminus is a CDK4/6-specific phosphorylation site pivotal for proteasomal degradation. During the late G1/S phase, phosphorylation at Ser672 by CDK4/6 marks p130 for recognition by the F-box protein Skp2, a component of the SCF^Skp2 ubiquitin ligase complex. This enables p130 polyubiquitination and rapid proteasomal turnover, independently of CDK2 activity. As a result, p130-mediated repression of E2F-responsive genes is relieved, allowing S-phase entry, DNA replication, and cell proliferation, while terminating the growth-suppressive function of p130. In quiescent G0 cells, hypophosphorylated p130 forms stable complexes with E2F4/5-DP1 to silence proliferation-associated genes; phosphorylation at Ser672 dynamically lowers p130 levels as cells re-enter the cycle. Dysregulation of the CDK4/6-Skp2-p130 axis or p130 mutations can stabilize hyperphosphorylated forms and disrupt cell cycle checkpoints, promoting uncontrolled proliferation and cancer development.

References
https://pubmed.ncbi.nlm.nih.gov/12435635/

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

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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%