CENPF Antibody [G20B19] | Primary Antibodies

Application: Reactivity: Human

Usage Information

Dilution
IHC1:1000 IF1:100

Application
IHC, IF

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
357 kDa

Datasheet & SDS

Biological Description

Specificity
CENPF Antibody [G20B19] detects endogenous levels of total CENPF protein.

Clone
G20B19

Synonym(s)
Centromere protein F, CENP-F, AH antigen, Kinetochore protein CENPF, Mitosin, CENPF

Background
Centromere protein F (CENPF, mitosin) is a large, cell‑cycle‑regulated nuclear matrix and kinetochore protein that functions as a dynamic scaffold coordinating chromosome alignment, kinetochore–microtubule attachment, spindle checkpoint signaling, and cell‑cycle progression at the G2/M transition. The polypeptide is predominantly coiled‑coil and forms dimers that accumulate on the nuclear matrix in late G2, then relocalize to outer kinetochores from prophase through early anaphase, before shifting to the spindle midzone and intercellular bridge in late anaphase and telophase, where it is subsequently degraded, reflecting tight temporal control linked to mitotic events. CENPF provides a docking platform at kinetochores for multiple microtubule‑associated and checkpoint proteins, including the motor CENP‑E and the dynein/LIS1/NDE1/NDEL1 complex, and contributes to recruitment or stabilization of Bub1 and related spindle checkpoint components, thereby promoting robust kinetochore–microtubule capture, chromosome congression, tension sensing, and proper activation and silencing of the spindle assembly checkpoint. Loss or silencing of CENPF weakens centromeric cohesion, delays or prevents stable bioriented attachments and metaphase plate formation, prolongs mitosis, and triggers persistent spindle checkpoint activation, leading to chromosome missegregation and aneuploidy, which identifies CENPF as a core factor for accurate chromosome segregation. CENPF interacts with DNA‑PK and contributes to interphase chromatin organization and DNA synthesis control, and its C‑terminal region modulates pocket protein (RB family) activity to influence cell‑cycle progression and lineage decisions, including roles in embryonic cardiomyocyte cycling, skeletal myogenesis, and cardiac lineage specification from embryonic stem cells. Additional interactions with syntaxin‑4 and SNAP25 connect CENPF to recycling vesicles and the microtubule network, implicating it in membrane trafficking and plasma‑membrane recycling during cell division. Expression of CENPF is low in quiescent cells and peaks in proliferating and mitotic cells, and high CENPF levels are observed in a broad range of tumors where they correlate with poor prognosis, high grade, and features of chromosomal instability, while functional studies in hepatocellular carcinoma and prostate cancer indicate that CENPF promotes G2/M transition, drives proliferation, and can modulate metabolic pathways such as pyruvate kinase M2 phosphorylation signaling.

Background

Centromere protein F (CENPF, mitosin) is a large, cell‑cycle‑regulated nuclear matrix and kinetochore protein that functions as a dynamic scaffold coordinating chromosome alignment, kinetochore–microtubule attachment, spindle checkpoint signaling, and cell‑cycle progression at the G2/M transition. The polypeptide is predominantly coiled‑coil and forms dimers that accumulate on the nuclear matrix in late G2, then relocalize to outer kinetochores from prophase through early anaphase, before shifting to the spindle midzone and intercellular bridge in late anaphase and telophase, where it is subsequently degraded, reflecting tight temporal control linked to mitotic events. CENPF provides a docking platform at kinetochores for multiple microtubule‑associated and checkpoint proteins, including the motor CENP‑E and the dynein/LIS1/NDE1/NDEL1 complex, and contributes to recruitment or stabilization of Bub1 and related spindle checkpoint components, thereby promoting robust kinetochore–microtubule capture, chromosome congression, tension sensing, and proper activation and silencing of the spindle assembly checkpoint. Loss or silencing of CENPF weakens centromeric cohesion, delays or prevents stable bioriented attachments and metaphase plate formation, prolongs mitosis, and triggers persistent spindle checkpoint activation, leading to chromosome missegregation and aneuploidy, which identifies CENPF as a core factor for accurate chromosome segregation. CENPF interacts with DNA‑PK and contributes to interphase chromatin organization and DNA synthesis control, and its C‑terminal region modulates pocket protein (RB family) activity to influence cell‑cycle progression and lineage decisions, including roles in embryonic cardiomyocyte cycling, skeletal myogenesis, and cardiac lineage specification from embryonic stem cells. Additional interactions with syntaxin‑4 and SNAP25 connect CENPF to recycling vesicles and the microtubule network, implicating it in membrane trafficking and plasma‑membrane recycling during cell division. Expression of CENPF is low in quiescent cells and peaks in proliferating and mitotic cells, and high CENPF levels are observed in a broad range of tumors where they correlate with poor prognosis, high grade, and features of chromosomal instability, while functional studies in hepatocellular carcinoma and prostate cancer indicate that CENPF promotes G2/M transition, drives proliferation, and can modulate metabolic pathways such as pyruvate kinase M2 phosphorylation signaling.

References
https://pubmed.ncbi.nlm.nih.gov/30526248/ https://pubmed.ncbi.nlm.nih.gov/23791740/

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%