All lanes : Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

Lane 1 : Rat brain lysates prepared in RIPA lysis method
Lane 2 : Rat brain lysates prepared in 1%SDS Hot lysis method

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 113 kDa

The lysates were prepared in 1%SDS Hot lysis method.

Blocking/diluting buffer & concentration: 5% NFDM/TBST

Observed MW: 112 kDa

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: PARP1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab191217 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab191217 was shown to specifically react with PARP1 when PARP1 knockout samples were used. Wild-type and PARP1 knockout samples were subjected to SDS-PAGE. ab191217 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.                        

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on epithelial cells and stromal cells of Human testis 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PARP1 with ab191217 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on HeLa cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows:-
-ve control 1: ab191217 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

All lanes : Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

Lane 1 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 1uM staurosporine for 4 hours whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution

Predicted band size: 113 kDa
Observed band size: 113,89 kDa why is the actual band size different from the predicted?


Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 1536009).

The lysates were prepared in 1%SDS Hot lysis method

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on epithelial cells and stromal cells of rat testis 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cells) cells labeling PARP1 with ab191217 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution (red).

The negative controls are as follows:-
-ve control 1: ab191217 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) at 1/1000 dilution.
-ve control 2:  Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

All lanes : Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysates
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cells) treated with 1uM staurosporine for 4 hours whole cell lysates

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 113 kDa
Observed band size: 113,55,89 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile observed is consistent with what has been described in the literature (PMID: 1536009).

The lysates were prepared in 1%SDS Hot lysis method

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling PARP1 with ab191217 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on epithelial cells and stromal cells of mouse testis 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) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

All lanes : Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

Lane 1 : Human fetal heart lysate
Lane 2 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/50000 dilution

Predicted band size: 113 kDa
Observed band size: 113 kDa


Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The lysates were prepared in 1%SDS Hot lysis method

Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution + Mouse heart lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/50000 dilution

Predicted band size: 113 kDa
Observed band size: 113 kDa


Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.

The lysates were prepared in 1%SDS Hot lysis method