Human NCK1 Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human NCK1 in direct ELISAs.
  • Source
    Monoclonal Mouse IgG2A Clone # 714506
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human NCK1
    Met1-Ser377
    Accession # P16333
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human, Mouse, and Rat NCK1 by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line, MOLT‑4 human acute lymphoblastic leukemia cell line, BaF3 mouse pro-B cell line, and Y3‑Ag rat myeloid cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human NCK1 Monoclonal Antibody (Catalog # MAB7008) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for NCK1 at approximately 47 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
NCK1 in MCF‑7 Human Cell Line. NCK1 was detected in immersion fixed MCF‑7 human breast cancer cell line using Mouse Anti-Human NCK1 Monoclonal Antibody (Catalog # MAB7008) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: NCK1

NCK1 (Non-catalytic region of tyrosine kinase 1; also Sh2/SH3 adaptor protein NCK-alpha ) is a 46-48 kDa member of the Nck family of adaptor proteins. NCK1 is ubiquitously expressed and may be found in both cytoplasm and nucleus. In the cytoplasm, it connects tyrosine kinases to actin reorganization which, in T cells, involves SLP-76 and VAV1. Upon DNA damage, NCK1 binds SOCS7 and is translocated into the nucleus where it participates in p53 phosphorylation and cell cycle arrest. Human NCK1 is 377 amino acids (aa) in length. It contains three N-terminal SH3 domains (aa 2-252) that mediate binding to Pro-rich regions of cytoplasmic proteins, and a C-terminal SH2 domain (aa 282-376) that binds to phosphorylated proteins or receptors. There are at least three utilized Ser, and one utilized Tyr phosphorylation sites. There are also three potential splice variants. One shows a 12 aa substitution for aa 1-76, a second possesses an 11 aa substitution for aa 77‑377, and a third contains a 37 aa substitution for aa 134-377. Full-length NCK1 shares 99% aa identity with mouse NCK1.

MAB7008 detects mouse and rat NCK1 in Western Blots. Cross-reactivity with mouse and rat samples in immunocytochemistry has not been tested.

  • Long Name:
    Non-catalytic Region of Tyrosine Kinase Adaptor Protein 1
  • Entrez Gene IDs:
    4690 (Human); 179731 (Mouse); 300955 (Rat)
  • Alternate Names:
    cytoplasmic protein NCK1; MGC12668; NCK adaptor protein 1melanoma NCK protein; NCK1; nck-1; NCKalpha; NCKNCK tyrosine kinase; non-catalytic region of tyrosine kinase; SH2/SH3 adaptor protein NCK-alpha
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Phosphoproteomics of collagen receptor networks reveals SHP-2 phosphorylation downstream of wild-type DDR2 and its lung cancer mutants.
    Authors: Iwai L, Payne L, Luczynski M, Chang F, Xu H, Clinton R, Paul A, Esposito E, Gridley S, Leitinger B, Naegle K, Huang P
    Biochem J, 2013;454(3):501-13.
    Species: Human
    Sample Type: Cell Culture Supernates
    Application: ELISA Capture
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Isotype Controls
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Mouse IgG2A Isotype Control

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Secondary Antibodies
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Mouse IgG HRP-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL557-conjugate Antibody

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Donkey Anti-Mouse IgG Biotinylated Antibody

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Mouse IgG HRP-conjugated Antibody

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Donkey Anti-Mouse IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL008 8
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Mouse F(ab)2 IgG (H+L) APC-conjugated Antibody

Flow F0101B 2  
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Donkey Anti-Mouse IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL009 5
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Mouse F(ab)2 IgG (H+L) Fluorescein-conjugated Antibody

Flow F0103B 5  
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Mouse IgG Antibody

WB AF007 4
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Goat Anti-Mouse IgG Biotinylated Antibody

WB BAF007 4
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Mouse/Rabbit IgG VisUCyte HRP Polymer Antibody

IHC VC002  
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Rat Anti-Mouse IgG2A PE-conjugated Antibody

Flow F0129 1  
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Mouse IgG VisUCyte HRP Polymer Antibody

IHC VC001  
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Mouse F(ab)2 IgG (H+L) PerCP-conjugated Antibody

Flow F0114  
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Goat Anti-Mouse IgG Fc Affinity Purified Polyclonal Ab

G-202-C 2
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Mouse IgG2A Antibody

Flow MAB0032  
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Rat Anti-Mouse IgG2A APC-conjugated Antibody

Flow F0130  
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Rat Anti-Mouse IgG2A PerCP-conjugated Antibody

Flow F0131  
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