Recombinant
RabMAb

Recombinant Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)

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Overview

  • Product name

    Anti-SESN2/Sestrin-2 antibody [EPR18907]
    See all SESN2/Sestrin-2 primary antibodies

  • Description

    Rabbit monoclonal [EPR18907] to SESN2/Sestrin-2

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human SESN2/Sestrin-2 aa 100 to the C-terminus. The exact sequence is proprietary.
    Database link: P58004

  • Positive control

    • WB: HeLa whole cell lysate treated with 10 mM H2O2 for 1 hour; LoVo, 293, NIH/3T3, HCT 116, Rat1, RAW 264.7, C6 and PC-12 whole cell lysates; Human colon, fetal liver, testis and fetal kidney lysates; Mouse spleen lysate. ICC/IF: HCT 116 cells. Flow Cyt: NIH/3T3 and HCT 116 cells. IP: HeLa whole cell lysate.

  • General notes

     

     

     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.

Properties

Applications

Our Abpromise guarantee covers the use of ab178518 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 1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
ICC/IF 1/100.

ICC/IF is recommended for human and rat only.
Flow Cyt 1/60.
IP 1/30.

Target

Images

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

    Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 10 mM H2O2 for 1 hour
    Lane 3 : LoVo (Human colorectal adenocarcinoma cell line) whole cell lysate
    Lane 4 : 293T (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 5 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Sestrin expression is induced by H2O2 treatment, which is consistent with what has been described in the literature (PMID: 25337554).

    Exposure time:3 minutes

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

    Lane 1 : Human colon lysate
    Lane 2 : Human fetal liver lysate
    Lane 3 : Human testis lysate
    Lane 4 : Human fetal kidney lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

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


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time:3 minutes

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

    Lane 1 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
    Lane 2 : Rat1 (Rat fibroblast cell line) whole cell lysate
    Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : Mouse spleen lysate
    Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1, 2, 3, 4 and 5: 3 minutes; Lane 6: 30 seconds.

  • Immunocytochemistry/ Immunofluorescence - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)
    Immunocytochemistry/ Immunofluorescence - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)

    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.

  • Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)
    Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)

    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.

  • Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)
    Flow Cytometry - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)

    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.

  • Immunoprecipitation - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)
    Immunoprecipitation - Anti-SESN2/Sestrin-2 antibody [EPR18907] (ab178518)

    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.

References

This product has been referenced in:

  • Carr M  et al. Upregulated expression of the antioxidant sestrin 2 identified by transcriptomic analysis of Japanese encephalitis virus-infected SH-SY5Y neuroblastoma cells. Virus Genes 55:630-642 (2019). Read more (PubMed: 31292858) »
  • Yang K  et al. Sestrin2 Suppresses Classically Activated Macrophages-Mediated Inflammatory Response in Myocardial Infarction through Inhibition of mTORC1 Signaling. Front Immunol 8:728 (2017). WB ; Mouse . Read more (PubMed: 28713369) »
See all 2 Publications for this product

Publishing research using ab178518? Please let us know so that we can cite the reference in this datasheet.

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