NEK7 Recombinant Rabbit Monoclonal Antibody [JE51-70]
Catalog# ET7110-16
NEK7 Recombinant Rabbit Monoclonal Antibody [JE51-70]
-
WB
-
IHC-P
-
FC
-
IF-Cell
-
Human
-
Mouse
-
Rat
-
unconjugated
Overview
Product Name
NEK7 Recombinant Rabbit Monoclonal Antibody [JE51-70]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within internal sequence of Human NEK7aa 200-302 / 302.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IHC-P, FC, IF-Cell
Molecular Weight
Predicted band size: 35 kDa
Positive Control
A549 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, Mouse lung cell lysate, C6 cell lysate, rat brain tissue lysate, A549, NIH/3T3, C6, human liver carcinoma tissue, mouse skeletal muscle tissue, human stomach tissue, rat brain tissue, Jurkat.
Conjugation
unconjugated
Clone Number
JE51-70
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:500-1:2,000
-
IHC-P
-
1:50-1:800
-
FC
-
1:50-1:100
-
IF-Cell
-
1:50-1:250
Target
Function
NIMA was originally shown in Aspergillus nidulans to be necessary for entry into mitosis. NIMA-related mammalian proteins have since been identified as Nek1-4 and Nek6-9. High expression of Nek1 is seen in male and female germ cell lines of mice. Nek2 is the closest known mammalian relative to NIMA. Like NIMA, Nek2 expression peaks at the G2 to M phase transition. Nek3, Nek6, Nek7 and Nek9 also regulate mitosis. Nek1 and Nek8 have been linked with polycystic kidney disease, and Nek4 expression is present in most primary carcinomas. Nek7 localizes to the cytoplasm and is highly expressed in lung, testis, muscle, slpeen, heart, liver, leukocyte and brain. Lower expression of Nek7 is detected in the ovary, prostate and kidney, while no expression is seen in small intestine.
Background References
1. Kim S. et. al. NEK7 is a centrosomal kinase critical for microtubule nucleation. Biochem. Biophys. Res. Commun. 360:56-62(2007).
2. O\'Regan L. et. al. The Nek6 and Nek7 protein kinases are required for robust mitotic spindle formation and cytokinesis. Mol. Cell. Biol. 29:3975-3990(2009).
Sequence Similarity
Belongs to the protein kinase superfamily. NEK Ser/Thr protein kinase family. NIMA subfamily.
Tissue Specificity
Highly expressed in lung, muscle, testis, brain, heart, liver, leukocyte and spleen. Lower expression in ovary, prostate and kidney. No expression seen in small intestine.
Post-translational Modification
Phosphorylation at Ser-195 required for its activation.
Subcellular Location
Cytoplasm, Cytoskeleton, Microtubule, Nucleus.
Synonyms
EC 2.7.11.1 antibody
NEK7 antibody
NEK7_HUMAN antibody
Never in mitosis A-related kinase 7 antibody
Never in mitosis gene a-related kinase 7 antibody
NIMA (never in mitosis gene a) related kinase 7 antibody
NIMA (never in mitosis gene a)-related expressed kinase 7 antibody
NimA related protein kinase 7 antibody
NIMA-related kinase 7 antibody
NimA-related protein kinase 7 antibody
ExpandImages
-
Western blot analysis of NEK7 on different lysates with Rabbit anti-NEK7 antibody (ET7110-16) at 1/1,000 dilution.
Lane 1: A549 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: NIH/3T3 cell lysate (20 µg/Lane)
Lane 4: Mouse lung tissue lysate (40 µg/Lane)
Lane 5: C6 cell lysate (no heat) (20 µg/Lane)
Lane 6: Rat brain tissue lysate (40 µg/Lane)
Notice: no heat means the lysate is not boiled.
Predicted band size: 35 kDa
Observed band size: 35 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7110-16) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of A549 cells labeling NEK7 with Rabbit anti-NEK7 antibody (ET7110-16) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NEK7 antibody (ET7110-16) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of NIH/3T3 cells labeling NEK7 with Rabbit anti-NEK7 antibody (ET7110-16) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NEK7 antibody (ET7110-16) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling NEK7 with Rabbit anti-NEK7 antibody (ET7110-16) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-NEK7 antibody (ET7110-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-NEK7 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-16, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue using anti-NEK7 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-16, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-NEK7 antibody (ET7110-16) at 1/800 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-16) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-NEK7 antibody (ET7110-16) at 1/800 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-16) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of Jurkat cells labeling NEK7.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET7110-16, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"