Nek2 Antibody [A24K3] | Primary Antibodies

Application: Reactivity: Human, Mouse, Rat

Usage Information

Dilution
WB1:100-1:1000 IP1:200-1:400 IHC1:50-1:500 IF1:50-1:500

Application
WB, IP, IHC, IF, ELISA

Reactivity
Human, Mouse, Rat

Source
Mouse 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
47 kDa

Positive Control	Human tonsil tissue; K-562 cells; A-431 cells; A549 cells; KNRK cells; NIH/3T3 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
Nek2 Antibody [A24K3] detects endogenous levels of total Nek2 protein.

Clone
A24K3

Synonym(s)
Serine/threonine-protein kinase Nek2; HSPK 21; Never in mitosis A-related kinase 2 (NimA-related protein kinase 2); NimA-like protein kinase 1; NEK2; NEK2A; NLK1

Background
Nek2, or NIMA-related kinase 2, is the closest mammalian homolog of the Aspergillus nidulans NIMA mitotic kinase and is a serine/threonine protein kinase that reaches peak activity at the G2/M transition. It is highly enriched at duplicated centrosomes, where it plays a critical role in regulating mitotic progression. Nek2 contains an N-terminal kinase domain with a bilobal catalytic core featuring conserved motifs and key residues such as aspartate 159 and aspartate 221 for catalysis, as well as autophosphorylation sites in the activation loop. It also has a central RCC1-like coiled-coil domain that mediates homodimerization and autoinhibition, and a C-terminal non-catalytic regulatory region. ADP-bound crystal structures of Nek2 reveal conformational flexibility in the nucleotide pocket and the existence of multiple kinase states. Nek2 phosphorylates centrosomal substrates, including C-Nap1, Rootletin, and gamma-tubulin, thereby triggering centrosome disjunction and splitting at the onset of prophase, which is essential for proper bipolar spindle formation. Nek2 also regulates kinetochore-microtubule attachments through phosphorylation of NDC80, mitotic checkpoint components such as CDC20 and MAD2L1, chromatin condensation via HMGA2, and the DNA damage response by activating ATM and ATR-CHEK1/2 pathways to delay mitotic entry. Nek2 coordinates the G2/M checkpoint with Wnt and beta-catenin signaling, influencing the expression of cyclin D1 and c-Myc, maintains genomic stability during embryonic development, and modulates splicing through phosphorylation of SRSF1, favoring anti-apoptotic isoforms. Inhibition of Nek2 leads to G2/M arrest and increased DNA damage and apoptosis, as seen in porcine embryos. Dysregulation of Nek2 drives oncogenesis, with overexpression observed in cancers such as hepatocellular carcinoma, breast cancer, and cervical cancer, where it promotes aneuploidy, and is also associated with neurodevelopmental defects.

Background

Nek2, or NIMA-related kinase 2, is the closest mammalian homolog of the Aspergillus nidulans NIMA mitotic kinase and is a serine/threonine protein kinase that reaches peak activity at the G2/M transition. It is highly enriched at duplicated centrosomes, where it plays a critical role in regulating mitotic progression. Nek2 contains an N-terminal kinase domain with a bilobal catalytic core featuring conserved motifs and key residues such as aspartate 159 and aspartate 221 for catalysis, as well as autophosphorylation sites in the activation loop. It also has a central RCC1-like coiled-coil domain that mediates homodimerization and autoinhibition, and a C-terminal non-catalytic regulatory region. ADP-bound crystal structures of Nek2 reveal conformational flexibility in the nucleotide pocket and the existence of multiple kinase states. Nek2 phosphorylates centrosomal substrates, including C-Nap1, Rootletin, and gamma-tubulin, thereby triggering centrosome disjunction and splitting at the onset of prophase, which is essential for proper bipolar spindle formation. Nek2 also regulates kinetochore-microtubule attachments through phosphorylation of NDC80, mitotic checkpoint components such as CDC20 and MAD2L1, chromatin condensation via HMGA2, and the DNA damage response by activating ATM and ATR-CHEK1/2 pathways to delay mitotic entry. Nek2 coordinates the G2/M checkpoint with Wnt and beta-catenin signaling, influencing the expression of cyclin D1 and c-Myc, maintains genomic stability during embryonic development, and modulates splicing through phosphorylation of SRSF1, favoring anti-apoptotic isoforms. Inhibition of Nek2 leads to G2/M arrest and increased DNA damage and apoptosis, as seen in porcine embryos. Dysregulation of Nek2 drives oncogenesis, with overexpression observed in cancers such as hepatocellular carcinoma, breast cancer, and cervical cancer, where it promotes aneuploidy, and is also associated with neurodevelopmental defects.

References
https://pubmed.ncbi.nlm.nih.gov/17197699/ https://pubmed.ncbi.nlm.nih.gov/12857871/

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%