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Rabbit Recombinant Monoclonal Nck antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples. Cited in 5 publications.


Images

Western blot - Anti-Nck antibody [Y531] (AB32120), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-Nck antibody [Y531] (AB32120), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-Nck antibody [Y531] (AB32120), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nck antibody [Y531] (AB32120), expandable thumbnail
  • Western blot - Anti-Nck antibody [Y531] (AB32120), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Expected
Expected
Rat
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species
Human
Dilution info
-
Notes

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

Expected
Expected

Species
Mouse
Dilution info
-
Notes

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

Predicted
Predicted

Species
Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/2000 - 1/10000
Notes

-

Expected
Expected

Species
Mouse
Dilution info
1/2000 - 1/10000
Notes

-

Predicted
Predicted

Species
Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/250
Notes

-

Expected
Expected

Species
Mouse
Dilution info
1/250
Notes

-

Predicted
Predicted

Species
Rat
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/1000
Notes

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

Expected
Expected

Species
Mouse
Dilution info
1/1000
Notes

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

Predicted
Predicted

Species
Rat
Dilution info
-
Notes

-

Target data

Function

Adapter protein which associates with tyrosine-phosphorylated growth factor receptors, such as KDR and PDGFRB, 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. Plays a role in ELK1-dependent transcriptional activation in response to activated Ras signaling. Modulates the activation of EIF2AK2/PKR by dsRNA. May play a role in cell adhesion and migration through interaction with ephrin receptors.

Alternative names

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

Recommended products

Rabbit Recombinant Monoclonal Nck antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse samples. Cited in 5 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
Y531
Purification technique
Affinity purification Protein A
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

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

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The Nck protein also referred to as Nck1 n c k or Nck is an adaptor protein integral in cell signaling pathways. It consists of a molecular mass around 45 kDa and exists mainly within the cytoplasm. The Nck proteins including Nck1 and its closely related form Nck2 are particularly prominent in cells of the immune and nervous systems. Functionally they facilitate interactions between receptor tyrosine kinases and downstream signaling molecules translating extracellular stimuli into cellular responses.

Biological function summary

The Nck proteins play significant roles in actin cytoskeleton reorganization influencing cell shape movement and adhesion. These proteins often participate as part of larger signaling complexes where they serve as scaffolds bringing together key molecules. Nck proteins directly interact with partners containing SH2 and SH3 domains which are important for various cellular dynamic processes. Their role in cytoskeletal dynamics aligns them with many cellular mechanisms including endocytosis and cell motility.

Pathways

Nck proteins significantly influence the regulation of the actin cytoskeleton pathway and the Rho GTPases pathway. Through these pathways they interact with proteins such as WASP and Rac regulating actin polymerization and cell migration. Nck’s ability to form complex signaling networks positions it as a central actor in transducing signals that affect cell shape and movement. It acts collaboratively with multiple signaling pathways to maintain cellular plasticity and response to extracellular signals.

Associated diseases and disorders

Nck proteins have associations with cancer and immunological conditions. Alterations in their signaling functions may contribute to tumor progression or metastasis given their role in cell movement and adhesion. In immune disorders abnormal Nck signaling could disrupt normal immune cell functions potentially leading to autoimmune conditions. For example Nck proteins may interact with oncogenic proteins like BCR-ABL in chronic myeloid leukemia highlighting their relevance in disease contexts.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

5 product images

  • Western blot - Anti-Nck antibody [Y531] (ab32120), expandable thumbnail

    Western blot - Anti-Nck antibody [Y531] (ab32120)

    Lanes 1-3: Merged signal (red and green). Green - ab32120 observed at 50 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human NCK1 (Nck) knockout HeLa cell line ab265323 (knockout cell lysate Human NCK1 (Nck) knockout HeLa cell lysate ab258059) was used. Wild-type and Nck knockout samples were subjected to SDS-PAGE. ab32120 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

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

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: NCK1 knockout HeLa cell lysate at 20 µg

    Lane 2: Western blot - Human NCK1 (Nck) knockout HeLa cell line (Human NCK1 (Nck) knockout HeLa cell line ab265323)

    Lane 3: NIH/3T3 cell lysate at 20 µg

    Secondary

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

    Predicted band size: 43 kDa

    Observed band size: 50 kDa

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

    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, Goat Anti-Rabbit IgG H&L (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), expandable thumbnail

    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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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), expandable thumbnail

    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), expandable thumbnail

    Western blot - Anti-Nck antibody [Y531] (ab32120)

    All lanes: Western blot - Anti-Nck antibody [Y531] (ab32120) at 1/10000 dilution

    All lanes: Hela cell lysate

    Predicted band size: 43 kDa

    Observed band size: 35 kDa, 47 kDa

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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