NCK1 Recombinant Rabbit Monoclonal Antibody [JU41-80]
Catalog# ET7106-57
NCK1 Recombinant Rabbit Monoclonal Antibody [JU41-80]
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
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unconjugated
Overview
Product Name
NCK1 Recombinant Rabbit Monoclonal Antibody [JU41-80]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human NCK1 aa 204-296 / 377.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Target Molecular Weight
43 kDa
Positive Control
Hela cell lysate, NIH/3T3 cell lysate, rat kidney tissue lysate, mouse testis tissue lysate, LOVO, A431, HUVEC, mouse testis tissue, rat testis tissue, human tonsil tissue, human kidney tissue, Jurkat.
Conjugation
unconjugated
Clone Number
JU41-80
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
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
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WB
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1:500-1:2,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
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IP
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1:10-1:50
Target
Function
The NCK family of SH2/SH3 adaptor proteins consists of two members, NCK1 (NCKα) and NCK2 (NCKβ), which couple tyrosine kinase signaling, including the EGF and PDGF receptor-pathways, to downstream signaling proteins. Specifically, overexpression of Nck1 in NIH/3T3 cells decreases DNA synthesis stimulated by EGF. Furthermore, the SH2 domain of NCK2 inhibits EGF- and PDGF-induced DNA synthesis. The SH3 domain of NCK binds a proline-rich domain on PAK, a known actin cytoskeleton regulator. The NCK protein thus mediates the interaction between PAK and RAC. The NCK2 protein binds human PDGFR-b (Tyr 1009); overexpression of Nck2 inhibits PDGF-induced membrane ruffling and lamellipod formation. Various growth factor receptors, cell surface antigens and adhesion molecules phosphorylate mammalian NCK1 and NCK2. The human NCK1 and NCK2 genes map to chromosomes 3q22.3 and 2q12.2, respectively.
Background References
1. Stein E et al. Nck recruitment to Eph receptor, EphB1/ELK, couples ligand activation to c-Jun kinase. J Biol Chem 273:1303-1308 (1998).
2. Braverman L E et al. Identification of Grb4/Nckbeta, a src homology 2 and 3 domain-containing adapter protein having similar binding and biological properties to Nck. J Biol Chem 274:5542-5549 (1999).
Post-translational Modification
Phosphorylated on Ser and Tyr residues. Phosphorylated in response to activation of EGFR and FcERI. Phosphorylated by activated PDGFRB.
Subcellular Location
Cytoplasm. Endoplasmic reticulum. Nucleus.
Synonyms
Cytoplasmic protein NCK1 antibody
Melanoma Nck protein antibody
MGC12668 antibody
NCK 1 antibody
NCK adaptor protein 1 antibody
NCK alpha antibody
NCK antibody
NCK tyrosine kinase antibody
Nck-1 antibody
NCK1 antibody
ExpandImages
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Western blot analysis of NCK1 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 (ET7106-57, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: Rat kidney tissue lysate
Lane 4: Mouse testis tissue lysate
Predicted band size: 43 kDa
Observed band size: 43/36 kDa -
ICC staining of NCK1 in LOVO cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of NCK1 in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of NCK1 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-57, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 1/50) 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.
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Immunohistochemical analysis of paraffin-embedded rat testis tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 1/50) 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.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 1/50) 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.
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Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-NCK1 antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-57, 1/50) 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.
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Flow cytometric analysis of NCK1 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7106-57, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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