NEK9 Antibody [E20D7] | Primary Antibodies

Application: Reactivity: Mouse, Rat, Human

Lane 1: A-431, Lane 2: A-431 (KO NEK9), Lane 3: Hep G2, Lane 4: NIH/3T3

Usage Information

Dilution
WB1:1000 - 1:10000 IHC1:50 - 1:100 IF1:100 FCM1:10 - 1:100

Application
WB, IHC, IF, FCM

Reactivity
Mouse, Rat, 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
107 kDa 120 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	Rat spleen; Human kidney tissue; Human testis tissue; NIH/3T3 cells; HeLa cells; THP-1 cells; A-431 cells; Hep G2 cells; MCF7 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
NEK9 Antibody [E20D7] detects endogenous levels of total NEK9 protein.

Clone
E20D7

Synonym(s)
KIAA1995, NEK8, NERCC, NEK9, Serine/threonine-protein kinase Nek9, Nercc1 kinase, Never in mitosis A-related kinase 9, NimA-related kinase 8, NimA-related protein kinase 9, Nek8

Background
NEK9 is a NIMA-related serine/threonine kinase that plays a central role in mitotic progression and centrosome dynamics, featuring an N-terminal kinase domain responsible for activating downstream kinases NEK6 and NEK7 through phosphorylation of their activation loops. Following the kinase domain, NEK9 contains seven RCC1-like β-propeller domains that serve as a scaffold for mediating diverse protein-protein interactions, and a C-terminal coiled-coil/regulatory region with a conserved GWLRKELENAEFIPMPDSP motif critical for binding and activating NEK7 by displacing its autoinhibitory Tyr97, as well as harboring an LC3-interacting region involved in autophagy regulation. NEK9 is activated at centrosomes during prophase by phosphorylation via PLK1 and CDK1, enabling it to phosphorylate targets such as NEDD1 at Ser377 to promote γ-tubulin recruitment for microtubule nucleation and TPX2 to regulate its localization prior to mitosis. NEK9 also orchestrates centrosome separation and bipolar spindle assembly through the NEK6/7-Eg5 pathway, and its C-terminal domain binds MYH9 to regulate actin dynamics crucial for ciliogenesis. Pathogenic mutations in NEK9 disrupt mitotic fidelity, leading to disorders such as microcephaly and Lethal Congenital Contracture Syndrome, while NEK9 overexpression is linked to tumorigenesis by promoting centrosome amplification and anaphase defects.

Background

NEK9 is a NIMA-related serine/threonine kinase that plays a central role in mitotic progression and centrosome dynamics, featuring an N-terminal kinase domain responsible for activating downstream kinases NEK6 and NEK7 through phosphorylation of their activation loops. Following the kinase domain, NEK9 contains seven RCC1-like β-propeller domains that serve as a scaffold for mediating diverse protein-protein interactions, and a C-terminal coiled-coil/regulatory region with a conserved GWLRKELENAEFIPMPDSP motif critical for binding and activating NEK7 by displacing its autoinhibitory Tyr97, as well as harboring an LC3-interacting region involved in autophagy regulation. NEK9 is activated at centrosomes during prophase by phosphorylation via PLK1 and CDK1, enabling it to phosphorylate targets such as NEDD1 at Ser377 to promote γ-tubulin recruitment for microtubule nucleation and TPX2 to regulate its localization prior to mitosis. NEK9 also orchestrates centrosome separation and bipolar spindle assembly through the NEK6/7-Eg5 pathway, and its C-terminal domain binds MYH9 to regulate actin dynamics crucial for ciliogenesis. Pathogenic mutations in NEK9 disrupt mitotic fidelity, leading to disorders such as microcephaly and Lethal Congenital Contracture Syndrome, while NEK9 overexpression is linked to tumorigenesis by promoting centrosome amplification and anaphase defects.

References
https://pubmed.ncbi.nlm.nih.gov/29250521/ https://pubmed.ncbi.nlm.nih.gov/34078910/

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%