重组Anti-PDE5A/PDE5抗体[EPR24129-95]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labelling PDE5A/PDE5 with ab259945 at 1/500 (0.928 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on artery smooth muscle of human brain.The section was incubated with ab259945 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling PDE5A/PDE5 with ab259945 at 1/500 (0.928 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : no staining on human liver.The section was incubated with ab259945 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labelling PDE5A/PDE5 with ab259945 at 1/500 (0.928 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on purkinje cells of human cerebellum.The section was incubated with ab259945 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

PDE5A/PDE5 was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with ab259945 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259945 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab259945 IP in MCF7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259945 in MCF7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 15 seconds

All lanes:

Immunoprecipitation - Anti-PDE5A/PDE5 antibody [EPR24129-95] (ab259945)

Predicted band size: 100 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of paraffin-embedded Rat cerebellum tissue labelling PDE5A/PDE5 with ab259945 at 1/500 (0.928 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on rat cerebellum.The section was incubated with ab259945 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of paraffin-embedded Mouse cerebellum tissue labelling PDE5A/PDE5 with ab259945 at 1/500 (0.928 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on mouse cerebellum.The section was incubated with ab259945 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cerebellum tissue labeling PDE5A/PDE5 with ab259945 at 1/100 (4.64 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution (Green). Positive staining on mouse cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat cerebellum tissue labeling PDE5A/PDE5 with ab259945 at 1/100 (4.64 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution (Green). Positive staining on rat cerebellum is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• WB

Lab

Western blot - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

Western blot : Anti-PDE5A antibody [EPR24129-95] (ab259945) 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, ab259945 was shown to bind specifically to PDE5A. A band was observed at 95 kDa in wild-type A549 cell lysates with no signal observed at this size in PDE5A knockout cell line. To generate this image, wild-type and PDE5A 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 3 % 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-PDE5A/PDE5 antibody [EPR24129-95] (ab259945) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human PDE5A knockout A549 cell line (<a href='/products/cell-lines/human-pde5a-knockout-a549-cell-line-ab301140'>ab301140</a>)

Lane 2:

PDE5A knockout A549 cell lysate at 20 µg

Lane 3:

K562 cell lysate at 20 µg

Secondary

Lanes 1 - 3:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 3:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 100 kDa

Observed band size: 95 kDa

false

• WB

Lab

Western blot - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

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

Negative control : liver (PMID : 11389965).

Exposure time : 37 seconds

All lanes:

Western blot - Anti-PDE5A/PDE5 antibody [EPR24129-95] (ab259945) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 40 µg

Lane 2:

Human liver tissue lysate at 40 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 100 kDa

false

• WB

Lab

Western blot - Anti-PDE5A/PDE5 antibody [EPR24129-95] (AB259945)

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

Negative control : liver (PMID : 11389965).

Exposure time : 37 seconds

All lanes:

Western blot - Anti-PDE5A/PDE5 antibody [EPR24129-95] (ab259945) at 1/1000 dilution

Lane 1:

Mouse esophagus tissue lysate at 40 µg

Lane 2:

Mouse cerebellum tissue lysate at 40 µg

Lane 3:

Mouse liver tissue lysate at 40 µg

Lane 4:

Rat cerebellum tissue lysate at 40 µg

Lane 5:

Rat liver tissue lysate at 40 µg

Secondary

Lanes 1 - 5:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 1 - 5:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/100000 dilution

Predicted band size: 100 kDa

false