Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) HeLa (human cervix adenocarcinoma epithelial cell, Right) cells labeling pan PDE4 with ab300108 at 1/5000 dilution (0.01ug) (red) compared with a Rabbit monoclonal IgG (ab172730) (black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody. Low expression : K-562.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling pan PDE4 with ab300108 at 1/500 (1.044 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab300108 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 Leica DS9800 (Bond™ Polymer Refine Detection) kit. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing cytoplasmic staining in HeLa cell line. Low expression : K-562. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5µg/ml) dilution (red). The Nuclear counterstain was DAPI (blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/mL) dilution (Green). Confocal image showing positive staining on rat cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab300108 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution (Green). Confocal image showing positive staining on mouse cerebrum. The nuclear counterstain was DAPI (Blue). The section was incubated with ab300108 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling pan PDE4 with ab300108 at 1/5000 (0.104 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on rat cerebrum. The section was incubated with ab300108 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 Leica DS9800 (Bond™ Polymer Refine Detection) kit used at a ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4µg/ml) dilution (red) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution. The Nuclear counterstain was DAPI (blue). -ve control : ab11267 at 1/500 (4μg/ml) followed by ab150081 at 1/1000 (2 ug/ml) dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron labeling pan PDE4 with ab300108 at 1/50 (10.44 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (green). Confocal image showing positive staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 (4µg/ml) dilution (red) followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution. The Nuclear counterstain was DAPI (blue). -ve control : ab11267 at 1/500 (4μg/ml) followed by ab150081 at 1/1000 (2 ug/ml) dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling pan PDE4 with ab300108 at 1/5000 (0.104 µg/ml) followed by a Leica DS9800 (Bond™ Polymer Refine Detection) kit. Positive staining on mouse cerebrum. The section was incubated with ab300108 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 Leica DS9800 (Bond™ Polymer Refine Detection) kit was used at a ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

• WB

Lab

Western blot - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. ab181602 was used as the Loading Control at 1/200000 dilution. Blocking/Diluting buffer and concentration : 5% NFDM/TBS Low expression : DU145 (PMID : 24518597), K562. The molecular weight observed is consistent with what has been described in the literature (PMID : 9880581, 15717866).

Lanes 1 - 7:

Western blot - Anti-pan PDE4 antibody [EPR25202-101] (<a href='/en-us/products/primary-antibodies/pan-pde4-antibody-epr25202-101-ab300108'>ab300108</a>) at 1/1000 dilution

Lanes 1 - 7:

Western blot - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (ab300109)

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 50 µg

Lane 3:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

Mouse brain tissue lysate at 50 µg

Lane 5:

Mouse cerebellum tissue lysate at 50 µg

Lane 6:

Rat brain tissue lysate at 50 µg

Lane 7:

Rat cerebellum tissue lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 60-100 kDa

false

• Dot

Lab

Dot Blot - Anti-pan PDE4 antibody [EPR25202-101] - BSA and Azide free (AB300109)

This data was developed using ab300108, the same antibody clone in a different buffer formulation. Dot blot analysis of pan PDE4 using ab300108 at 1/1000 (0.522 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100,000 dilution. Exposure time : 8 seconds Blocking and diluting buffer and concentration : 5% NFDM/TBST Lane 1 : His-tagged human PDE4A fragment Lane 2 : His-tagged human PDE4B fragment Lane 3 : GST-tagged human PDE4C fragment Lane 4 : His-tagged human PDE4D fragment