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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

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

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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)
  • Immunocytochemistry/ Immunofluorescence - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
  • Western blot - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
  • Immunoprecipitation - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
  • Flow Cytometry (Intracellular) - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
  • Flow Cytometry (Intracellular) - Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free (ab236025)
  • 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: Flow Cyt (Intra), IP, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 PE Unconjugated

製品の概要

  • 製品名

    Anti-SESN2/Sestrin-2 antibody [EPR18907] - BSA and Azide free
    SESN2/Sestrin-2 一次抗体 製品一覧

  • 製品の詳細

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

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), IP, ICC/IF, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

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

  • ポジティブ･コントロール

    • ICC/IF: HCT 116 cells. WB: HeLa, LoVo and HEK-293T, HEK-293 cell lysate. Flow Cyt (intra): NIH/3T3 and HCT 116 cells. IP: HeLa cell lysate.

  • 特記事項

    ab236025 is the carrier-free version of ab178518.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab236025の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

ICC/IF is recommended for human and rat only.
WB
Use at an assay dependent concentration. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
特記事項
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

ICC/IF is recommended for human and rat only.
WB
Use at an assay dependent concentration. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).

ターゲット情報

画像

  • 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 HEK-293 cell lysate
    Lane 2 : SESN2 knockout HEK-293 cell lysate

    Performed under reducing conditions.

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



    False colour image of Western blot: Anti-SESN2/Sestrin-2 antibody [EPR18907] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab178518 was shown to bind specifically to SESN2/Sestrin-2. A band was observed at 54 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in SESN2 knockout cell line ab269486 (knockout cell lysate ab269650). To generate this image, wild-type and SESN2 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

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

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

    Intracellular 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)

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (2)

ab236025 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab236025 は 2 報の論文で使用されています。

  • Wang H  et al. Sestrin2 Is Increased in Calcific Aortic Disease and Inhibits Osteoblastic Differentiation in Valvular Interstitial Cells via the Nuclear Factor E2-related Factor 2 Pathway. J Cardiovasc Pharmacol 80:609-615 (2022). PubMed: 35881902
  • Keping Y  et al. Sestrin1 inhibits oxidized low-density lipoprotein-induced activation of NLRP3 inflammasome in macrophages in a murine atherosclerosis model. Eur J Immunol 50:1154-1166 (2020). PubMed: 32297666

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