Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-LAG-3 antibody [EPR20294-77] (ab209238)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20294-77] to LAG-3
  • Suitable for: WB, ICC/IF, IP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-LAG-3 antibody [EPR20294-77]
    See all LAG-3 primary antibodies

  • Description

    Rabbit monoclonal [EPR20294-77] to LAG-3

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    ICC/IF
    Mouse
    IP
    Mouse
    WB
    Mouse
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus; mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours, whole cell lysate. ICC/IF: Mouse splenocytes. IP: Mouse splenocytes treated with 5 µg/ml concanavalin A for 72 hours, whole cell lysate.

  • General notes

    This product was previously labelled as Lymphocyte Activation Gene 3.

    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.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Immunocytochemistry/ Immunofluorescence - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized untreated and concanavalin A (2.5 μg/ml for 72 hours) treated mouse splenocytes labeling LAG-3 with ab209238 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing increased membranous staining following treatment of mouse splenocytes with concanavalin A (2.5 μg/ml for 72 hours) (PMID 20391435; PMID 26318293; PMID 11527700; PMID 19631993).

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution

  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/10000 dilution + Mouse LAG-3 recombinant protein (aa24-442) tagged with murine IgG2A at the C-terminus, 10 ng

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking/Dilution Buffer: 5% NFDM/TBST

     

  • Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Western blot - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    All lanes : Anti-LAG-3 antibody [EPR20294-77] (ab209238) at 1/1000 dilution

    Lane 1 : Untreated mouse splenocytes whole cell lysate
    Lane 2 : Mouse splenocytes treated with 5 ug/ml concanavalin A for 72 hours whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 57 kDa
    Observed band size: 54, 70 kDa why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking/Dilution Buffer: 5% NFDM/TBST

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    LAG3 expression can be elevated in activated splenocytes (PMID 19631993).

    The molecular mass observed is consistent with the literature (PMID 15971272; PMID 15557174).

  • Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Immunoprecipitation - Anti-LAG-3 antibody [EPR20294-77] (ab209238)

    LAG-3 was immunoprecipitated from 0.35 mg of mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours) whole cell lysate with ab209238 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209238 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate 10μg (Input). 

    Lane 2: ab209238 IP in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209238 in mouse splenocytes (treated with 5 μg/ml concanavalin A for 72 hours), whole cell lysate.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 1 second.

  • Anti-LAG-3 antibody [EPR20294-77] (ab209238)
    Anti-LAG-3 antibody [EPR20294-77] (ab209238)

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

Alternative products to Anti-LAG-3 antibody [EPR20294-77] (ab209238)

© 2021 Astana Biomed Group LLP. All rights reserved.