Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18907] to SESN2/Sestrin-2 - BSA and Azide free
  • Suitable for: IP, ICC/IF, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free
    See all SESN2/Sestrin-2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18907] to SESN2/Sestrin-2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, Flow Cyt, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • ICC/IF: HCT 116 cells. WB: HeLa, LoVo and HEK293T cell lysate. Flow Cyt: NIH/3T3 and HCT 116 cells. IP: HeLa cell lysate.

  • General notes

    ab236025 is the carrier-free version of ab178518 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab236025 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product was previously labelled as SESN2

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SESN2 knockout HeLa cell lysate
    Lane 3 : LoVo cell lysate
    Lane 4 : HEK-293T 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

    Predicted band size: 54 kDa
    Observed band size: 54 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab178518).

    Lanes 1-4: Merged signal (red and green). Green - ab178518 observed at 54 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab178518 Anti-SESN2/Sestrin-2 antibody [EPR18907] was shown to specifically react with 2 in wild-type HeLa cells. Loss of signal was observed when knockout sample ab257665 was used. Wild-type and 2 knockout samples were subjected to SDS-PAGE. ab178518 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-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Immunocytochemistry/ Immunofluorescence - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (Human colorectal carcinoma cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HCT116 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)  at 1/250 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178518).

  • Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    All lanes : Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : SESN2 knockout HeLa cell lysate
    Lane 3 : HeLa 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

    Predicted band size: 54 kDa
    Observed band size: 54 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab178518).

    Lanes 1-3: Merged signal (red and green). Green - ab178518 observed at 54 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab178518 Anti-SESN2/Sestrin-2 antibody [EPR18907] was shown to specifically react with SESN2/Sestrin-2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265669 (knockout cell lysate ab257665) was used.  Wild-type and SESN2/Sestrin-2 knockout samples were subjected to SDS-PAGE.  ab178518 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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.

  • Immunoprecipitation - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Immunoprecipitation - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

    SESN2/Sestrin-2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab178518 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab178518 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate, 10µg (Input).

    Lane 2: ab178518 IP in HeLa whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab178518 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    SESN2/Sestrin-2 expression is low in HeLa cells and can be enriched through immunoprecipitation.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178518).

  • Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

    Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal[EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178518).

  • Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

    Flow cytometric analysis of 4% paraformaldehyde-fixed HCT 116 (Human colorectal carcinoma cell line) cells labeling SESN2/Sestrin-2 with ab178518 at 1/60 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178518).

  • Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
    Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)

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