Nck-2 Recombinant Rabbit Monoclonal Antibody [JE63-36]
Catalog# HA722573
Nck-2 Recombinant Rabbit Monoclonal Antibody [JE63-36]
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WB
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IF-Cell
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IHC-P
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IF-Tissue
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Human
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unconjugated
Overview
Product Name
Nck-2 Recombinant Rabbit Monoclonal Antibody [JE63-36]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Nck-2 aa 51-100 / 380.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P, IF-Tissue
Molecular Weight
Predicted band size: 43 kDa
Positive Control
A431 cell lysate, SK-MEL-28 cell lysate, HDLM-2 cell lysate, Caco-2 cell lysate, THP-1 cell lysate, A431, human testis tissue.
Conjugation
unconjugated
Clone Number
JE63-36
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
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WB
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1:1,000
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IF-Cell
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1:100
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IHC-P
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1:1,000
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IF-Tissue
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1:200
Target
Function
Cytoplasmic protein NCK2 (also known as NCK-beta and Grb4) is a protein that in humans is encoded by the NCK2 gene. NCK belongs to family of adaptor proteins. There are two mammalian NCK genes, NCK1 and NCK2. NCK1 is located in chromosome 3 and NCK2 is located in chromosome 2. The protein contains three SH3 domains and one SH2 domain. The protein has no known catalytic function but has been shown to bind and recruit various proteins involved in the regulation of receptor protein tyrosine kinases. It is through these regulatory activities that this protein is believed to be involved in cytoskeletal reorganization. Alternate transcriptional splice variants, encoding different isoforms, have been characterized.
Background References
1. Bywaters BC et al. Endothelial NCK2 promotes atherosclerosis progression in male but not female Nck1-null atheroprone mice. Front Cardiovasc Med. 2022 Aug
2. Cui Y et al. OIP5 Interacts with NCK2 to Mediate Human Spermatogonial Stem Cell Self-Renewal and Apoptosis through Cell Cyclins and Cycle Progression and Its Abnormality Is Correlated with Male Infertility. Research (Wash D C). 2023 Jun
Subcellular Location
Cytoplasm, Endoplasmic reticulum.
UNIPROT
Synonyms
Cytoplasmic protein NCK2 antibody
GRB4 antibody
Growth factor receptor bound protein 4 antibody
NCK adaptor protein 2 antibody
Nck-2 antibody
NCK2 antibody
NCK2_HUMAN antibody
NCKbeta antibody
Non-catalytic region of tyrosine kinase adaptor protein 2 antibody
Noncatalytic region of tyrosine kinase beta antibody
ExpandImages
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Western blot analysis of Nck-2 on different lysates with Rabbit anti-Nck-2 antibody (HA722573) at 1/1,000 dilution.
Lane 1: A431 cell lysate
Lane 2: SK-MEL-28 cell lysate
Lane 3: HDLM-2 cell lysate
Lane 4: Caco-2 cell lysate
Lane 5: THP-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 43 kDa
Observed band size: 46 kDa
Exposure time: 1 minute 34 seconds; 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 (HA722573) 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 A431 cells labeling Nck-2 with Rabbit anti-Nck-2 antibody (HA722573) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Nck-2 antibody (HA722573) 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 testis tissue with Rabbit anti-Nck-2 antibody (HA722573) at 1/1,000 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 (HA722573) at 1/1,000 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.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"