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Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)

Key features and details

  • Mouse monoclonal [1F3] to SIRT1 - Nuclear Marker
  • Suitable for: WB, IHC-P, Flow Cyt, ICC/IF
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-SIRT1 antibody [1F3] - Nuclear Marker
    See all SIRT1 primary antibodies

  • Description

    Mouse monoclonal [1F3] to SIRT1 - Nuclear Marker

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment corresponding to Human SIRT1.
    Database link: Q96EB6

  • Positive control

    • WB: MDA-MB-231, MCF7, Jurkat, HeLa, HEK293 and A549 cell lysates. IHC-P: Human lung and kidney cancer tissue. ICC: NTERA-2 cells. Flow Cyt: K562 cells.

  • General notes

    This product was changed from ascites to supernatant. Lot no’s high than GR255594-13 are from Tissue Culture Supernatant

Properties

Images

  • Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    All lanes : Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833) at 1/500 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: 120 kDa
    Observed band size: 110 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab104833 observed at 110 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.

    ab104833 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 ab104833 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    ab104833 at 1/200 dilution staining SIRT1 in Human lung cancer by Immunohistochemistry, Paraffin-embedded tissue with DAB staining.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    ab104833 at 1/200 dilution staining SIRT1 in Human kidney cancer by Immunohistochemistry, Paraffin-embedded tissue with DAB staining.
  • Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Immunocytochemistry/ Immunofluorescence - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    ab104833 at 1/200 dilution staining SIRT1 in NTERA-2 cells by Immunofluorescence (green). Red: Actin filaments have been labeled with Alexa Fluor-555 phalloidin.
  • Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    All lanes : Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833) at 1/500 dilution

    Lane 1 : MCF7 cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : HEK293 cell lysate
    Lane 5 : A549 cell lysate

    Predicted band size: 120 kDa

  • Flow Cytometry - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Flow Cytometry - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    ab104833 staining SIRT1 in K562 cells by Flow Cytometry shown in green, negative control in purple.
  • Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833)
    Western blot - Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833) This image is courtesy of an anonymous Abreview
    Anti-SIRT1 antibody [1F3] - Nuclear Marker (ab104833) at 1/1000 dilution + Rat smooth muscle cells at 100000 cells

    Secondary
    HRP-conjugated Sheep anti-mouse polyclonal IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 120 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 1 hour

    See Abreview

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