Gankyrin Antibody (Rabbit mAb) [K15B19]

Application: Reactivity: Human

Usage Information

Dilution
WB1:1000 - 1:10000 IHC1:50 - 1:100 IF1:100 FCM1:80

Application
WB, IHC, IF, FCM

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
24 kDa 24 kDa
^*Why do the predicted and actual molecular weights differ? The following reasons may explain differences between the predicted and actual protein molecular weight. Post-translational modifications(e.g., phosphorylation, glycosylation); Splice variants and isoforms; Relative charge; Multimerization.

Datasheet & SDS

Biological Description

Specificity
Gankyrin Antibody (Rabbit mAb) [K15B19] detects endogenous levels of total Gankyrin protein.

Clone
K15B19

Synonym(s)
26S proteasome non-ATPase regulatory subunit 10, 26S proteasome regulatory subunit p28, Gankyrin, p28(GANK), PSMD10

Background
Gankyrin, also known as PSMD10 or p28GANK, is a proteasome-associated oncoprotein belonging to the ankyrin repeat family, acting as a regulatory subunit of the 26S proteasome that integrates cell-cycle, stress-response, and oncogenic signaling through high-affinity protein–protein interactions mediated by a tandem array of ankyrin repeats. The ankyrin repeat architecture provides a modular scaffold that engages cyclin-dependent kinase 4 (CDK4), the retinoblastoma tumor suppressor (RB), and the E3 ubiquitin ligase MDM2, positioning gankyrin as a central adaptor that couples phosphorylation, ubiquitination, and proteasomal degradation of key cell-cycle and apoptotic regulators. Within the CDK4–RB axis, gankyrin binding promotes CDK4 activation and enhances RB phosphorylation, followed by accelerated recruitment of phosphorylated RB to the 26S proteasome, which diminishes RB-mediated repression of E2F transcription factors and supports sustained entry into and progression through the cell cycle in proliferating hepatocytes and tumor cells. In parallel, interaction with MDM2 increases MDM2 E3 ligase activity toward p53, augmenting p53 ubiquitination and delivery of polyubiquitylated p53 to the proteasome, thereby reducing p53-dependent transcriptional programs linked to DNA damage responses and apoptosis and favoring survival under genotoxic and oncogenic stress. Gankyrin facilitates the targeting function of the proteasome by bridging CDK4 and MDM2 with the 19S regulatory particle, which supports more efficient recognition and processing of multiple polyubiquitylated substrates and consolidates its role as a dual negative regulator of the RB and p53 tumor suppressor pathways. In hepatocellular carcinoma (HCC), gankyrin is frequently overexpressed and extends its regulatory impact to metabolic signaling by activating β-catenin/c-Myc signaling, leading to increased transcription of glucose and glutamine transporters and glycolytic and glutaminolytic enzymes, such as GLUT1 and GLS1, which collectively shift cellular metabolism toward enhanced aerobic glycolysis and glutaminolysis. This metabolic reprogramming supports anabolic growth, migration, and metastatic potential and associates high gankyrin expression with increased glucose uptake, lactate production, glutamine utilization, and glutamate synthesis in HCC cell lines and patient-derived xenografts, indicating that gankyrin coordinates proteasome-dependent degradation of tumor suppressors with c-Myc-driven metabolic adaptation. The interplay between gankyrin and β-catenin also affects transcriptional circuits beyond metabolism, as elevated β-catenin correlates with high gankyrin levels in HCC biopsies and the combined expression pattern predicts poor overall survival, consistent with its wider impact on Wnt/β-catenin target genes involved in proliferation and invasion. Overexpression of gankyrin in liver cancer cells enhances proliferation, protects against apoptosis induced by DNA-damaging agents, and promotes tumorigenicity and metastasis, outcomes that reflect simultaneous attenuation of RB and p53 signaling and reinforcement of oncogenic β-catenin/c-Myc pathways. Inhibition of gankyrin-mediated proteasomal interactions reduces CDK4 and MDM2 activity, restores RB and p53 protein levels, and increases apoptosis in cancer cells, underscoring its importance as a mechanistic hub that governs tumor suppressor stability and cellular fate decisions. At the structural-functional level, the ankyrin repeat array and proteasome-binding surfaces enable gankyrin to operate as a docking platform that coordinates substrate recognition, ubiquitin-tag transfer, and proteasome engagement, properties that are directly relevant when designing assays to monitor gankyrin activity or screening for small molecules that disrupt its interactions with CDK4, MDM2, or the 19S regulatory complex. In physiological contexts such as liver regeneration and chronic liver disease, gankyrin-driven degradation of tumor suppressor proteins contributes to hepatocyte proliferation and may facilitate the transition from compensatory growth to malignant transformation when its expression becomes chronically elevated. Dysregulation of gankyrin, particularly in HCC and other gastrointestinal malignancies, is therefore characterized by coordinated suppression of RB and p53 checkpoints and reinforcement of β-catenin/c-Myc metabolic signaling, a combination that underpins early cell-cycle deregulation, sustained anabolic metabolism, and enhanced metastatic behavior and provides multiple mechanistic entry points for therapeutic targeting.

Background

Gankyrin, also known as PSMD10 or p28GANK, is a proteasome-associated oncoprotein belonging to the ankyrin repeat family, acting as a regulatory subunit of the 26S proteasome that integrates cell-cycle, stress-response, and oncogenic signaling through high-affinity protein–protein interactions mediated by a tandem array of ankyrin repeats. The ankyrin repeat architecture provides a modular scaffold that engages cyclin-dependent kinase 4 (CDK4), the retinoblastoma tumor suppressor (RB), and the E3 ubiquitin ligase MDM2, positioning gankyrin as a central adaptor that couples phosphorylation, ubiquitination, and proteasomal degradation of key cell-cycle and apoptotic regulators. Within the CDK4–RB axis, gankyrin binding promotes CDK4 activation and enhances RB phosphorylation, followed by accelerated recruitment of phosphorylated RB to the 26S proteasome, which diminishes RB-mediated repression of E2F transcription factors and supports sustained entry into and progression through the cell cycle in proliferating hepatocytes and tumor cells. In parallel, interaction with MDM2 increases MDM2 E3 ligase activity toward p53, augmenting p53 ubiquitination and delivery of polyubiquitylated p53 to the proteasome, thereby reducing p53-dependent transcriptional programs linked to DNA damage responses and apoptosis and favoring survival under genotoxic and oncogenic stress. Gankyrin facilitates the targeting function of the proteasome by bridging CDK4 and MDM2 with the 19S regulatory particle, which supports more efficient recognition and processing of multiple polyubiquitylated substrates and consolidates its role as a dual negative regulator of the RB and p53 tumor suppressor pathways. In hepatocellular carcinoma (HCC), gankyrin is frequently overexpressed and extends its regulatory impact to metabolic signaling by activating β-catenin/c-Myc signaling, leading to increased transcription of glucose and glutamine transporters and glycolytic and glutaminolytic enzymes, such as GLUT1 and GLS1, which collectively shift cellular metabolism toward enhanced aerobic glycolysis and glutaminolysis. This metabolic reprogramming supports anabolic growth, migration, and metastatic potential and associates high gankyrin expression with increased glucose uptake, lactate production, glutamine utilization, and glutamate synthesis in HCC cell lines and patient-derived xenografts, indicating that gankyrin coordinates proteasome-dependent degradation of tumor suppressors with c-Myc-driven metabolic adaptation. The interplay between gankyrin and β-catenin also affects transcriptional circuits beyond metabolism, as elevated β-catenin correlates with high gankyrin levels in HCC biopsies and the combined expression pattern predicts poor overall survival, consistent with its wider impact on Wnt/β-catenin target genes involved in proliferation and invasion. Overexpression of gankyrin in liver cancer cells enhances proliferation, protects against apoptosis induced by DNA-damaging agents, and promotes tumorigenicity and metastasis, outcomes that reflect simultaneous attenuation of RB and p53 signaling and reinforcement of oncogenic β-catenin/c-Myc pathways. Inhibition of gankyrin-mediated proteasomal interactions reduces CDK4 and MDM2 activity, restores RB and p53 protein levels, and increases apoptosis in cancer cells, underscoring its importance as a mechanistic hub that governs tumor suppressor stability and cellular fate decisions. At the structural-functional level, the ankyrin repeat array and proteasome-binding surfaces enable gankyrin to operate as a docking platform that coordinates substrate recognition, ubiquitin-tag transfer, and proteasome engagement, properties that are directly relevant when designing assays to monitor gankyrin activity or screening for small molecules that disrupt its interactions with CDK4, MDM2, or the 19S regulatory complex. In physiological contexts such as liver regeneration and chronic liver disease, gankyrin-driven degradation of tumor suppressor proteins contributes to hepatocyte proliferation and may facilitate the transition from compensatory growth to malignant transformation when its expression becomes chronically elevated. Dysregulation of gankyrin, particularly in HCC and other gastrointestinal malignancies, is therefore characterized by coordinated suppression of RB and p53 checkpoints and reinforcement of β-catenin/c-Myc metabolic signaling, a combination that underpins early cell-cycle deregulation, sustained anabolic metabolism, and enhanced metastatic behavior and provides multiple mechanistic entry points for therapeutic targeting.

References
https://pubmed.ncbi.nlm.nih.gov/16177571/ https://pubmed.ncbi.nlm.nih.gov/30503555/

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%