NCK1 Polyclonal antibody

• Featured Product
• KD/KO Validated

NCK1 Polyclonal Antibody for WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), IP, ELISA

Conjugate

Unconjugated

Publications

5

Cat no : 15247-1-AP

Synonyms

SH2/SH3 adapter protein NCK-alpha, SH2/SH3 adapter protein NCK1, NCKalpha, Nck-1, NCK alpha

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Validation Data Gallery

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  WB analysis using 15247-1-AP

  Various lysates were subjected to SDS PAGE followed by western blot with 15247-1-AP (NCK1 antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse brain using 15247-1-AP

  mouse brain tissue were subjected to SDS PAGE followed by western blot with 15247-1-AP (NCK1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse liver using 15247-1-AP

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 15247-1-AP (NCK1 antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of rat brain using 15247-1-AP

  rat brain tissue were subjected to SDS PAGE followed by western blot with 15247-1-AP (NCK1 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours.

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  IP experiment of mouse brain using 15247-1-AP

  IP result of anti-NCK1 (IP:15247-1-AP, 4ug; Detection:15247-1-AP 1:500) with mouse brain tissue lysate 2640ug.

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  IHC staining of human breast cancer using 15247-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15247-1-AP (NCK1 antibody) at dilution of 1:200 (under 10x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human breast cancer using 15247-1-AP

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 15247-1-AP (NCK1 antibody) at dilution of 1:200 (under 40x lens. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 15247-1-AP

  Immunofluorescent analysis of Hela cells, using NCK1 antibody 15247-1-AP at 1:25 dilution and Rhodamine-labeled goat anti-rabbit IgG (red).

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  FC experiment of HeLa using 15247-1-AP

  1x10^6 HeLa cells were intracellularly stained with 0.4 ug NCK1 Polyclonal antibody (15247-1-AP) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.4 ug Rabbit IgG control Rabbit PolyAb (30000-0-AP) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: mouse brain tissue, mouse liver tissue, rat brain tissue
• Positive IP detected in: mouse brain tissue
• Positive IHC detected in: human breast cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: Hela cells
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:10-1:100
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 3 publications below
• WB: See 5 publications below
• IHC: See 1 publications below
• IP: See 1 publications below

Product Information

15247-1-AP targets NCK1 in WB, IHC, IF/ICC, FC (Intra), IP, ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: NCK1 fusion protein Ag0979
• Full Name: NCK adaptor protein 1
• Calculated Molecular Weight: 43 kDa
• Observed Molecular Weight: 43 kDa
• GenBank Accession Number: BC006403
• Gene Symbol: NCK1
• Gene ID (NCBI): 4690
• RRID: AB_2150428
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

NCK family adaptor proteins function to couple tyrosine phosphorylation signals to regulate actin cytoskeletal reorganization that leads to cell motility. Cytoplasmic protein NCK1 (or NCK alpha) is an adapter protein which associates with tyrosine-phosphorylated growth factor receptors, such as KDR and PDGFRB, or their cellular substrates. NCK1 binds a number of intracellular proteins and influences various signaling pathways including GTPase-activating protein of Ras (RasGAP), which is responsible for the down-regulation of Ras. NCK1 is recently reported to regulate the UPR, which secondary to obesity impairs glucose homeostasis and INS actions.

Publications

Species	Application	Title
human	WB	Cell Signal; Identification of NCK1 as a novel downstream effector of STAT3 in colorectal cancer metastasis and angiogenesis.; Authors - Fan Zhang ; KD Validated
human	WB,IP	Mol Immunol; The adapter proteins ADAP and Nck cooperate in T cell adhesion.; Authors - Marcus Lettau ; KD Validated
human	WB,IHC	Mol Biol Rep; STAT3-induced NCK1 elevation promotes migration of triple-negative breast cancer cells via regulating ERK1/2 signaling; Authors - Peina He ; KD Validated
human	WB	Cancer Biomark; Long noncoding RNA LINC00885 upregulates NCK1 to promote cell viability and migration of triple-negative breast cancer cells through sponging miR-654-3p; Authors - Peina He
human	WB	Cell Microbiol; The Legionella pneumophila effector WipA disrupts host F-actin polymerisation by hijacking phosphotyrosine signalling.; Authors - Lei He