NEK9 Polyclonal antibody

NEK9 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

2

Cat no : 11192-1-AP

Synonyms

NimA-related kinase 8, Never in mitosis A-related kinase 9, NERCC, Nek8, KIAA1995

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Validation Data Gallery

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  WB analysis of HeLa using 11192-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 11192-1-AP (NEK9 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours.

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  WB analysis using 11192-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 11192-1-AP (NEK9 antibody) at dilution of 1:4500 incubated at room temperature for 1.5 hours.

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  WB analysis of human placenta using 11192-1-AP

  human placenta tissue were subjected to SDS PAGE followed by western blot with 11192-1-AP (NEK9 antibody) at dilution of 1:4500 incubated at room temperature for 1.5 hours.

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  IP experiment of HeLa using 11192-1-AP

  IP result of anti-NEK9 (IP:11192-1-AP, 4ug; Detection:11192-1-AP 1:1000) with HeLa cells lysate 1880 ug.

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  IHC staining of human lymphoma using 11192-1-AP

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 11192-1-AP (NEK9 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human lymphoma using 11192-1-AP

  Immunohistochemical analysis of paraffin-embedded human lymphoma tissue slide using 11192-1-AP (NEK9 antibody) at dilution of 1:200 (under 0x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HepG2 using 11192-1-AP

  Immunofluorescent analysis of HepG2 cells, using NEK9 antibody 11192-1-AP at 1:50 dilution and FITC-labeled donkey anti-rabbit IgG(green). Blue pseudocolor = DAPI (fluorescent DNA dye).

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  IF Staining of HepG2 using 11192-1-AP

  Immunofluorescent analysis of (4% PFA) fixed HepG2 cells using NEK9 antibody (11192-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L).

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  FC experiment of HeLa using 11192-1-AP

  1X10^6 HeLa cells were intracellularly stained with 0.4 ug Anti-Human NEK9 (11192-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) at dilution 1:1000 (red), or 0.4 ug x. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HeLa cells, human placenta tissue, HepG2 cells
• Positive IP detected in: HeLa cells
• Positive IHC detected in: human lymphoma tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HepG2 cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:9000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below
• IF: See 1 publications below

Product Information

11192-1-AP targets NEK9 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NEK9 fusion protein Ag1678
• Full Name: NIMA (never in mitosis gene a)- related kinase 9
• Calculated Molecular Weight: 107 kDa
• Observed Molecular Weight: 120 kDa
• GenBank Accession Number: BC009336
• Gene Symbol: NEK9
• Gene ID (NCBI): 91754
• RRID: AB_2150800
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NEK9 is also named as KIAA1995, NEK8, NERCC and belongs to the NEK Ser/Thr protein kinase family. It is originally identified as αβ-casein kinase that associates with a putative substrate Bicd2. It is also independently identified as a Nek6- and Ran GTPase-binding protein under a different name, Nercc1(PMID:14660563). Nek9, together with the highly similar (80% identical) Nek6 and Nek7, form a signaling module that is activated during mitosis and involved in the regulation of the mitotic spindle. The endogenous Nek9 (with predicted molecular mass of 120 kDa) has an apparent molecular mass of 600 kDa, also compatible with a tetramer(PMID:21454704).

Publications

Species	Application	Title
human	IF	JBMR Plus; Identification of Chloride Intracellular Channel Protein 3 as a Novel Gene Affecting Human Bone Formation.; Authors - Andrea M Brum
human	WB	Cancers (Basel); Differential Expression of NEK Kinase Family Members in Esophageal Adenocarcinoma and Barrett's Esophagus; Authors - Lei Chen