For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/primary-antibodies/nck-antibody-y531-ab32120.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Assembly
Share by email
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Nck antibody [Y531] (ab32120)

  • Datasheet
  • SDS
Reviews (1) Submit a question References (5)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-Nck antibody [Y531] (ab32120)
  • Immunocytochemistry/ Immunofluorescence - Anti-Nck antibody [Y531] (ab32120)
  • Flow Cytometry (Intracellular) - Anti-Nck antibody [Y531] (ab32120)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nck antibody [Y531] (ab32120)
  • Western blot - Anti-Nck antibody [Y531] (ab32120)
  • Anti-Nck antibody [Y531] (ab32120)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y531] to Nck
  • Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Protein
Recombinant Human Nck protein (ab172816)
Knockout
Product image
Human NCK1 (Nck) knockout HeLa cell line (ab265323)

View more associated products

Overview

  • Product name

    Anti-Nck antibody [Y531]
    See all Nck primary antibodies
  • Description

    Rabbit monoclonal [Y531] to Nck
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Hela cell lysate and human breast carcinoma ICC/IF: HeLa cells
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y531
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Assembly
    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Proto-oncogenes
    • Cancer
    • Signal transduction
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Other

Associated products

  • Alternative Versions

    • Anti-Nck antibody [Y531] - BSA and Azide free (ab235389)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human NCK1 (Nck) knockout HeLa cell line (ab265323)
  • KO cell lysates

    • Human NCK1 (Nck) knockout HeLa cell lysate (ab258059)
  • Recombinant Protein

    • Recombinant Human Nck protein (ab172816)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32120 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB
1/2000 - 1/10000. Detects a band of approximately 47 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/250.
Notes
WB
1/2000 - 1/10000. Detects a band of approximately 47 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF
1/250.

Target

  • Function

    Adapter protein which associates with tyrosine-phosphorylated growth factor receptors or their cellular substrates. Maintains low levels of EIF2S1 phosphorylation by promoting its dephosphorylation by PP1. Plays a role in the DNA damage response, not in the detection of the damage by ATM/ATR, but for efficient activation of downstream effectors, such as that of CHEK2.
  • Sequence similarities

    Contains 1 SH2 domain.
    Contains 3 SH3 domains.
  • Post-translational
    modifications

    Phosphorylated on Ser and Tyr residues.
  • Cellular localization

    Cytoplasm. Endoplasmic reticulum. Nucleus. Mostly cytoplasmic, but shuttles between the cytoplasm and the nucleus. Import into the nucleus requires the interaction with SOCS7. Predominantly nuclear following genotoxic stresses, such as UV irradiation, hydroxyurea or mitomycin C treatments.
  • Target information above from: UniProt accession P16333 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4690 Human
    • Entrez Gene: 17973 Mouse
    • Entrez Gene: 300955 Rat
    • Omim: 600508 Human
    • SwissProt: P16333 Human
    • SwissProt: Q99M51 Mouse
    • Unigene: 477693 Human
    • Alternative names

      • 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
      • NCK1_HUMAN antibody
      • NCKalpha antibody
      • Non catalytic region of tyrosine kinase antibody
      • SH2/SH3 adaptor protein NCK alpha antibody
      • SH2/SH3 adaptor protein NCK-alpha antibody
      see all

    Images

    • Western blot - Anti-Nck antibody [Y531] (ab32120)
      Western blot - Anti-Nck antibody [Y531] (ab32120)
      All lanes : Anti-Nck antibody [Y531] (ab32120) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : NCK1 knockout HeLa cell lysate
      Lane 3 : NIH/3T3 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Observed band size: 50 kDa why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab32120 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab32120 Anti-Nck antibody [Y531] was shown to specifically react with Nck in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265323 (knockout cell lysate ab258059) was used.  Wild-type and Nck knockout samples were subjected to SDS-PAGE.  ab32120 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Nck antibody [Y531] (ab32120)
      Immunocytochemistry/ Immunofluorescence - Anti-Nck antibody [Y531] (ab32120)

      Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Nck with purified ab32120 at 1/250 dilution (8 µg/mL). Cells were fixed in 100% Methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain.

    • Flow Cytometry (Intracellular) - Anti-Nck antibody [Y531] (ab32120)
      Flow Cytometry (Intracellular) - Anti-Nck antibody [Y531] (ab32120)

      Overlay histogram showing Jurkat cells stained with ab32120 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32120, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nck antibody [Y531] (ab32120)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nck antibody [Y531] (ab32120)

      ab32120 (1/100), staining human breast carcinoma by immunohistochemistry, Paraffin embedded tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Western blot - Anti-Nck antibody [Y531] (ab32120)
      Western blot - Anti-Nck antibody [Y531] (ab32120)
      Anti-Nck antibody [Y531] (ab32120) at 1/10000 dilution + Hela cell lysate

      Observed band size: 47 kDa why is the actual band size different from the predicted?
      Additional bands at: 35 kDa. We are unsure as to the identity of these extra bands.

    • Anti-Nck antibody [Y531] (ab32120)
      Anti-Nck antibody [Y531] (ab32120)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (5)

    Publishing research using ab32120? Please let us know so that we can cite the reference in this datasheet.

    ab32120 has been referenced in 5 publications.

    • Guan T  et al. Aristolochic acid inhibits Slit2-induced migration and tube formation via inactivation of Robo1/Robo2-NCK1/NCK2 signaling pathway in human umbilical vein endothelial cells. Toxicol Lett 300:51-58 (2019). PubMed: 30381256
    • Xia P  et al. NCK1 promotes the angiogenesis of cervical squamous carcinoma via Rac1/PAK1/MMP2 signal pathway. Gynecol Oncol N/A:N/A (2018). PubMed: 30442385
    • Wu X  et al. Activation of diverse signalling pathways by oncogenic PIK3CA mutations. Nat Commun 5:4961 (2014). PubMed: 25247763
    • He B  et al. A remote cis-acting variant at 3q links glomerular NCK1 to diabetic nephropathy. PLoS One 8:e56414 (2013). WB ; Mouse . PubMed: 23441190
    • de Castro RO  et al. Once phosphorylated, tyrosines in carboxyl terminus of protein-tyrosine kinase Syk interact with signaling proteins, including TULA-2, a negative regulator of mast cell degranulation. J Biol Chem 287:8194-204 (2012). WB . PubMed: 22267732

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunohistochemistry (Frozen sections) abreview for Anti-Nck antibody [Y531]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (spinal cord, motor neuron)
    Permeabilization
    No
    Specification
    spinal cord, motor neuron
    Blocking step
    Serum as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted May 16 2016

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

    Get resources and offers direct to your inbox Sign up
    A-Z by research area
    • Cancer
    • Cardiovascular
    • Cell biology
    • Developmental biology
    • Epigenetics & Nuclear signaling
    • Immunology
    • Metabolism
    • Microbiology
    • Neuroscience
    • Signal transduction
    • Stem cells
    A-Z by product type
    • Primary antibodies
    • Secondary antibodies
    • Biochemicals
    • Isotype controls
    • Flow cytometry multi-color selector
    • Kits
    • Loading controls
    • Lysates
    • Peptides
    • Proteins
    • Slides
    • Tags and cell markers
    • Tools & Reagents
    Help & support
    • Support
    • Make an Inquiry
    • Protocols & troubleshooting
    • Placing an order
    • RabMAb products
    • Biochemical product FAQs
    • Training
    • Browse by Target
    Company
    • Corporate site
    • Investor relations
    • Company news
    • Careers
    • About us
    • Blog
    Events
    • Tradeshows
    • Conferences
    International websites
    • abcam.cn
    • abcam.co.jp

    Join with us

    • LinkedIn
    • facebook
    • Twitter
    • YouTube
    • Terms of sale
    • Website terms of use
    • Cookie policy
    • Privacy policy
    • Legal
    • Modern slavery statement
    © 1998-2023 Abcam plc. All rights reserved.