Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Lck antibody [Y123] (ab32149)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y123] to Lck
  • Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
  • Reacts with: Human

Overview

  • Product name

    Anti-Lck antibody [Y123]
    See all Lck primary antibodies

  • Description

    Rabbit monoclonal [Y123] to Lck

  • Host species

    Rabbit

  • Specificity

    This antibody is specific for human Lck, it does not cross react with any other SRC family members. WB results were negative for the following Mouse and Rat tissues/cell lines: Ms brain, heart, kidney, spleen, C6, Raw264.7, PC12, NIH3T3, Ms thymus and serum.

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human Lck aa 1-100. The exact sequence is proprietary.

  • Epitope

    ab32149 reacts with an epitope located in the region before SH3 domain of Lck.

  • Positive control

    • Jurkat cell lysate, human lymphoma, HuT-78, BxPC-3 whole cell lysates.

  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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

Images

  • Western blot - Anti-Lck antibody [Y123] (ab32149)
    Western blot - Anti-Lck antibody [Y123] (ab32149)
    All lanes : Anti-Lck antibody [Y123] (ab32149) at 1/5000 dilution (purified)

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 2 : HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates
    Lane 3 : Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 58 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunoprecipitation - Anti-Lck antibody [Y123] (ab32149)
    Immunoprecipitation - Anti-Lck antibody [Y123] (ab32149)

    ab32149 (purified) at 1:20 dilution (2ug) immunoprecipitating Lck in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate.
    Lane 1 (input): Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10ug
    Lane 2 (+): ab32149 & Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32149 in Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry - Anti-Lck antibody [Y123] (ab32149)
    Flow Cytometry - Anti-Lck antibody [Y123] (ab32149)

    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Lck with purified ab32149 at 1:40 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilized with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Western blot - Anti-Lck antibody [Y123] (ab32149)
    Western blot - Anti-Lck antibody [Y123] (ab32149)
    Anti-Lck antibody [Y123] (ab32149) at 1/1000 dilution (purified) + BxPC-3 (Human pancreas adenocarcinoma epithelial cell) whole cell lysates at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 58 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [Y123] (ab32149)
    Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [Y123] (ab32149)

    Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling Lck with purified ab32149 at 1:500 dilution (0.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] (ab32149)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] (ab32149)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Hodgkin lymphoma tissue sections labeling Lck with Purified ab32149 at 1:250 dilution (1.5 μg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] (ab32149)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [Y123] (ab32149)

    Unpurified ab32149 at a 1/250 dilution staining Lck in human lymphoma using Immunohistochemistry, Paraffin Embedded Tissue.

  • Flow Cytometry - Anti-Lck antibody [Y123] (ab32149)
    Flow Cytometry - Anti-Lck antibody [Y123] (ab32149)

    Overlay histogram showing Jurkat cells stained with unpurified ab32149 (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 (ab32149, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). 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.

  • Western blot - Anti-Lck antibody [Y123] (ab32149)
    Western blot - Anti-Lck antibody [Y123] (ab32149)
    Anti-Lck antibody [Y123] (ab32149) at 1/1000 dilution (unpurified) + Jurkat cell lysate.

    Predicted band size: 58 kDa
    Observed band size: 58 kDa

  • Anti-Lck antibody [Y123] (ab32149)
    Anti-Lck antibody [Y123] (ab32149)

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