Anti-SIRT1 antibody [19A7AB4 ] - Nuclear Marker (ab110304)

  Immunocytochemistry - SIRT1 antibody [19A7AB4 ] (ab110304)

Immunocytochemistry analysis using ab110304 at 0.5µg/ml staining SIRT1 in HDFn cells (paraformaldehyde fixed and Triton X-100 permeabilized). The secondary antibody was (Red) Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps.

  Western blot - SIRT1 antibody [19A7AB4 ] (ab110304)

All lanes : Anti-SIRT1 antibody [19A7AB4 ] - Nuclear Marker (ab110304) at 0.125 µg/ml

Lane 1 : HepG2 cell lysate(Human)
Lane 2 : H9C2 cell lysate (Rat)
Lane 3 : MEF cell lysate (Mouse)


Predicted band size : 81 kDa


ab110304 detects a band of approximately 110 kDa (110-121 kDa) which is likely to be due to post translational glycosylation. SIRT1 is known to bind to several other proteins, and the 121kDa band could also be due to the presence of one of these complexes (ensure samples are adequately reduced and denatured).

  Flow Cytometry - SIRT1 antibody [19A7AB4 ] (ab110304)

HL-60 cells were stained with 1 µg/mL SIRT1 antibody ab110304(blue) or an equal amount of an isotype control antibody (red) and analyzed by flow cytometry.

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT1 antibody [19A7AB4 ] (ab110304)

IHC image of SIRT1 staining in Human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110304, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times

  Western blot - Anti-SIRT1 antibody [19A7AB4 ] - Nuclear Marker (ab110304)

Anti-SIRT1 antibody [19A7AB4 ] - Nuclear Marker (ab110304) at 1/8000 dilution + Mouse liver tissue lysate at 30 µg

Secondary
HRP-conjugated Goat anti-Mouse polyclonal IgG at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 81 kDa
Observed band size : 110 kDa (why is the actual band size different from the predicted?)


Exposure time : 30 seconds

This image is courtesy of an anonymous Abreview

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