Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18239] to SIRT1 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SIRT1 antibody [EPR18239] - BSA and Azide free
    See all SIRT1 primary antibodies

  • Description

    Rabbit monoclonal [EPR18239] to SIRT1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    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 testis tissue lysate; HEK293, MDA-MB-231, F9, HeLa and A549 whole cell lysates; Rat E18 brain tissue lysate. IHC-P: Human testis and skeletal muscle tissue; Mouse testis tissue; Rat skeletal muscle tissue. ICC/IF: HeLa and F9 cells. Flow cyt: HeLa and F9 cells. IP: F9 whole cell lysate.

  • General notes

    Ab233398 is the carrier-free version of ab189494. 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

    ab233398 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Western blot - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    All lanes : Anti-SIRT1 antibody [EPR18239] (ab189494) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293 cell lysate
    Lane 2 : SIRT1 knockout HEK-293 cell lysate
    Lane 3 : MDA-MB-231 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 80 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab189494).

    Lanes 1 - 4: Merged signal (red and green). Green - ab189494 observed at 110 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab189494 was shown to react with SIRT1 in western blot. The band observed in the knockout lysate lane below 110kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab189494 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Flow Cytometry - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized F9 (mouse embryonic testicular cancer cell line) cell line labeling SIRT1 with ab189494 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 H&L (Alexa Fluor® 488) (ab150077), 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 (ab189494).

  • Flow Cytometry - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Flow Cytometry - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling SIRT1 with ab189494 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 H&L (Alexa Fluor® 488) (ab150077), 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 (ab189494).

  • Immunoprecipitation - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunoprecipitation - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    SIRT1 was immunoprecipitated from 0.35 mg of F9 (mouse embryonic testicular cancer cell line) whole cell lysate with ab189494 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab189494 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: F9 whole lysate 10 μg (Input).
    Lane 2: ab189494 IP in F9 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab189494 in F9 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 50 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189494).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunohistochemical analysis of paraffin-embedded rat skeletal muscle tissue labeling SIRT1 with ab189494 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining in rat skeletal muscle (PMID: 23332867) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using Citrate, pH 6.0.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189494).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling SIRT1 with ab189494 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Mainly nuclear staining in mouse testis (PMID: 17197703) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using Citrate, pH 6.0.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189494).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunohistochemical analysis of paraffin-embedded human testis tissue labeling SIRT1 with ab189494 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Mainly nuclear staining in human testis (PMID: 17197703) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using Citrate, pH 6.0.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189494).

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% methanol-permeabilized F9 (mouse embryonic testicular cancer cell line) cells labeling SIRT1 with ab189494 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 nuclear and weakly cytoplasmic staining in F9 cell line.

    The nuclear counterstain 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) 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 (ab189494).

  • Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling SIRT1 with ab189494 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 nuclear and weakly cytoplasmic staining in HeLa cell line.

    The nuclear counterstain 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) 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 (ab189494).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

    Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling SIRT1 with ab189494 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining in human skeletal muscle (PMID: 23332867) is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

    Perform heat-mediated antigen retrieval using Citrate, pH 6.0.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189494).

  • Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)
    Anti-SIRT1 antibody [EPR18239] - BSA and Azide free (ab233398)

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