Western blot - PARP antibody (ab6079)

All lanes : Anti-PARP antibody (ab6079) at 1/250 dilution

Lane 1 : 30ug untreated HeLa whole cell lysate
Lane 2 : 30ug HeLa whole cell lysate treated with 1 uM Staurosporine overnight

Secondary
HRP conjugated Goat anti-rabbit

Predicted band size : 122 kDa
Observed band size : 29 kDa (why is the actual band size different from the predicted?)

This image is courtesy of an Abreview submitted by Dr Jerome Lemonnier

Immunocytochemistry/ Immunofluorescence - PARP antibody (ab6079)

ab6079 (1/500) staining PARP in assynchronous HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilised with 0.5% Triton X-100 and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see abreview.

Image part of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-PARP antibody(ab6079)

Ab6079 staining Human temporal cortex. Staining is localized to the nucleus.
Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

Western blot - PARP antibody (ab6079)

All lanes : Anti-PARP antibody (ab6079) at 1/1000 dilution

Lane 1 : 15ug mouse MEF cell lysate (untreated)
Lane 2 : 15ug mouse MEF cell lysate: cells treated for 1 hour with adriamycin
Lane 3 : 15ug mouse MEF cell lysate: cells treated for 8 hours with adriamycin

Secondary
HRP conjugated donkey anti-rabbit polyclonal antibody
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 122 kDa
Observed band size : 113 kDa (why is the actual band size different from the predicted?)


Exposure time : 1 minute

This image is courtesy of an Abreview submitted by Zeng Li.

Western blot - PARP antibody (ab6079)

All lanes : Anti-PARP antibody (ab6079) at 1/400 dilution

Lane 1 : Lysate prepared from human HeLa cytosolic extracts
Lane 2 : Lysate prepared from human HeLa nuclear extracts

Lysates/proteins at 50 µg per lane.

Secondary
HRP-conjugated goat polyclonal to rabbit IgG at 1/5000 dilution
developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 122 kDa
Observed band size : 29,112 kDa (why is the actual band size different from the predicted?)


Exposure time : 15 seconds

This image is a courtesy of Anonymous Abreview

Immunocytochemistry/ Immunofluorescence - Anti-PARP antibody (ab6079)

ICC/IF image of ab6079 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab6079 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.