Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon carcinoma. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized U-87 MG (human glioblastoma-astrocytoma epithelial cell) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml)(Green). Confocal image showing cytoplasmic staining in of U-87 MG cell line. Negative control : MCF7 (PMID : 18199541) is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml)(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 dilution (2 μg/ml).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon tissue labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling COX2 / Cyclooxygenase 2 with ab283574 at 1/500 dilution (1.058 μg/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on human colon liver. The section was incubated with ab283574 overnight at 4°C. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)

• IP

Supplier Data

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

COX2 / Cyclooxygenase 2 was immunoprecipitated from 0.35 mg U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate with ab283574 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283574 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate 10μg

Lane 2 : ab283574 IP in U-87 MG whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283574 in U-87 MG whole cell lysate

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

Exposure time : 6 seconds

Lower bands could be COX-2 fragments due to proteolysis. (PMID : 32366045)

All lanes:

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-rm1026-ab283574'>ab283574</a>)

Predicted band size: 69 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling COX2 / Cyclooxygenase 2 with ab283574 at 1/1000 dilution (0.529 μg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (2 μg/ml)(Green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line after treatment with lipopolysaccharide (1 μg/ml) for 6 hours is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 μg/ml)(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 dilution (2 μg/ml).

• IP

Supplier Data

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

COX2 / Cyclooxygenase 2 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1 μg/ml LPS for 6h whole cell lysate with ab283574 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283574 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) treated with 1ug/ml LPS for 6h whole cell lysate 10μg

Lane 2 : ab283574 IP in RAW 264.7 treated with 1 μg/ml LPS for 6h whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283574 in RAW 264.7 treated with 1 μg/ml LPS for 6h whole cell lysate

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

Exposure time : 6 seconds

All lanes:

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-rm1026-ab283574'>ab283574</a>)

Predicted band size: 69 kDa

false

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] - BSA and Azide free (AB283593)

This data was developed using ab283574, the same antibody clone in a different buffer formulation.

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

Negative control : MCF7 (PMID : 24325753, PMID : 16997132)

Lower bands could be COX-2 fragments due to proteolysis. (PMID : 32366045)

Exposure time : Lane 1-4 : 2 min Lane 5-6 : 3 min

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [RM1026] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-rm1026-ab283574'>ab283574</a>) at 1/1000 dilution

Lane 1:

U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 4:

RAW 264.7 treated with 1 µg/ml lipopolysaccharide (LPS) for 6h whole cell lysate at 20 µg

Lane 5:

C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 6:

C6 treated with 100 ng/ml lipopolysaccharide (LPS) for 4h whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 69 kDa

false