重组Anti-PARP1抗体[EPR18461]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

• WB

Supplier Data

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

Blocking/Dilution buffer : 5% NFDM/TBST.

The lysates were prepared in 1%SDS Hot lysis method

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

Lane 1:

Human fetal heart lysate at 10 µg

Lane 2:

Human fetal kidney lysate at 10 µg

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

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

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

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

Lane 1:

Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates at 10 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) treated with 1uM staurosporine for 4 hours whole cell lysates at 10 µg

Secondary

All lanes:

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

Predicted band size: 113 kDa

Observed band size: 113 kDa,89 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

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

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

Observed MW : 112 kDa

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

Lane 1:

Rat brain lysates prepared in RIPA lysis method at 15 µg

Lane 2:

Rat brain lysates prepared in 1%SDS Hot lysis method at 15 µg

Secondary

All lanes:

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

Predicted band size: 113 kDa

false

• WB

Lab

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

Western blot : Anti-PARP1 antibody [EPR18461] (ab191217) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab191217 was shown to bind specifically to PARP1. A band was observed at 125 kDa in wild-type A549 cell lysates with no signal observed at this size in PARP1 knockout cell line ab276094 (knockout cell lysate Abcam Pools). To generate this image, wild-type and PARP1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

Lane 1:

Wild-type A549 control staurosporine (0 uM, 72 h) cell lysate at 20 µg

Lane 2:

Wild-type A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

Lane 3:

PARP1 knockout A549 control staurosporine (0 uM, 72 h) cell lysate at 20 µg

Lane 4:

PARP1 knockout A549 treated staurosporine (3 uM, 24 h) cell lysate at 20 µg

Lane 5:

Empty cell lysate at 20 µg

Lane 6:

HAP1 Treated Staurosporine (2uM, 4h) cell lysate at 20 µg

Lane 7:

HAP1 Vehicle Control Staurosporine (0uM, 4h) cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

false

• WB

Supplier Data

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

Blocking/Dilution buffer : 5% NFDM/TBST.

The lysates were prepared in 1%SDS Hot lysis method

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/1000 dilution

All lanes:

Mouse heart lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 113 kDa

Observed band size: 113 kDa

false

Exposure time: 1min

• WB

Lab

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

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.

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217)

Predicted band size: 113 kDa

false

• WB

Supplier Data

Western blot - Anti-PARP1 antibody [EPR18461] (AB191217)

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

All lanes:

Western blot - Anti-PARP1 antibody [EPR18461] (ab191217) at 1/10000 dilution

Lane 1:

Untreated NIH/3T3 (Mouse embryonic fibroblast cells) whole cell lysates at 10 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cells) treated with 1uM staurosporine for 4 hours whole cell lysates at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 113 kDa

Observed band size: 113 kDa,55 kDa,89 kDa

false

Exposure time: 3min