NEK7 Recombinant Rabbit Monoclonal Antibody [JE51-70]
cat.: ET7110-16
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, IF-Cell
Clonality: Monoclonal
Clone number: JE51-70
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 35 kDa
Isotype: IgG
Immunogen: Recombinant protein within internal sequence of Human NEK7aa 200-302 / 302.
Positive control: NIH/3T3 cell lysate, Jurkat cell lysate, A549, NIH/3T3, C6, rat brain tissue, human liver carcinoma tissue, human stomach tissue, mouse skeletal muscle tissue.
Subcellular location: Cytoplasm, Cytoskeleton, Microtubule, Nucleus.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IF-Cell
1:500-1:2,000
1:50-1:800
1:50-1:100
1:50-1:250
Uniprot #: SwissProt: Q8TDX7 Human | Q9ES74 Mouse | D3ZBE5 Rat
Alternative names: EC 2.7.11.1 NEK7 NEK7_HUMAN Never in mitosis A-related kinase 7 Never in mitosis gene a-related kinase 7 NIMA (never in mitosis gene a) related kinase 7 NIMA (never in mitosis gene a)-related expressed kinase 7 NimA related protein kinase 7 NIMA-related kinase 7 NimA-related protein kinase 7 Serine/threonine-protein kinase Nek7
Images
ET7110-16_1.jpg Fig1: Western blot analysis of NEK7 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-16, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: NIH/3T3 cell lysate
Lane 2: Jurkat cell lysate
ET7110-16_2.jpg Fig2: 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.
ET7110-16_3.jpg Fig3: 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.
ET7110-16_4.jpg Fig4: 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.
ET7110-16_5.jpg Fig5: 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) for 20 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.
ET7110-16_6.jpg Fig6: 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) for 20 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.
ET7110-16_7.jpg Fig7: 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.
ET7110-16_8.jpg Fig8: 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.
ET7110-16_9.jpg Fig9: 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
Lane 2: Jurkat cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: Mouse lung cell lysate
Lane 5: C6 cell lysate (no heat)
Lane 6: Rat brain tissue lysate

Cell lysates/proteins at 10 µg/Lane.
Tissue lysates/proteins at 10 µg/Lane.

Predicted band size: 35 kDa
Observed band size: 35 kDa

Exposure time: 60 seconds;

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.
ET7110-16_10.jpg Fig10: Flow cytometric analysis of NEK7 was done on A549 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-16, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET7110-16_11.jpg Fig11: 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).
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