Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

This data was developed using ab179800, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling COX2/ Cyclooxygenase 2 with ab179800 at a concentration of 0.01µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab179800 Anti-COX2/Cyclooxygenase 2 antibody [EPR12012] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human colon tissue labeling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

This IHC data was generated using the same anti-COX2 / Cyclooxygenase 2 antibody clone, EPR12012, in a different buffer formulation (cat# ab179800).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling COX2 with unpurified ab179800 at a dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded) analysis of human liver tissue labelling COX2 / Cyclooxygenase 2 with unpurified ab179800 at a dilution of 1/250.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Western blot analysis on immunoprecipitation pellet from A549 cell lysate using unpurified ab179800.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

All lanes:

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>)

Predicted band size: 69 kDa

false

• IP

Lab

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

ab179800 (purified) at 1/30 immunoprecipitating COX2 in A549 whole cell lysate.

Lane 1 (input) : A549 whole cell lysate (10µg)

Lane 2 (+) : ab179800 + A549 whole cell lysate.

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab179800 in A549 whole cell lysate.

For western blotting, HRP-conjugated anti-rabbit IgG, specific for the reduced form of IgG, was used as the secondary antibody (1/1500).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

All lanes:

Immunoprecipitation - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>)

Predicted band size: 69 kDa

Observed band size: 79 kDa

false

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : secondary antibody only control. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling COX2 / Cyclooxygenase 2 with purified ab179800 at 1/4000 dilution (0.125 µg/ml).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : secondary antibody only control. Hematoxylin was used as a counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab179800)

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Western blot : Anti-PTGS2 antibody [EPR12012] (ab179800) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab179800 was shown to bind specifically to PTGS2. A band was observed at 69 kDa in wild-type RAW 264.7 cell lysates with no signal observed at this size in PTGS2 knockout cell line. To generate this image, wild-type and PTGS2 knockout RAW 264.7 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. This data was developed using the same antibody clone in a different buffer formulation (ab179800).

Lanes 1 - 4:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (ab227528)

Lane 1:

RAW 264.7 Control LPS (0 ng/mL, 4 h) cell lysate at 20 µg

Lane 2:

RAW 264.7 Treated LPS (100 ng/mL, 4 h) cell lysate at 20 µg

Lane 3:

Wild-type A549 ab277305 cell lysate at 20 µg

Lane 4:

PTGS2 knockout A549 ab283802 cell lysate at 20 µg

Observed band size: 69 kDa

false

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Exposure time : lane 1 : 180 seconds, lane 2 : 140 seconds. We recommend using a higher sensitive ECL substrate to increase the band intensity This data was developed using the same antibody clone in a different buffer formulation (ab179800).

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

All lanes:

A549 (Human lung carcinoma epithelial cell) 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 1/20000 dilution

false

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

False colour image of Western blot : Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab179800 was shown to bind specifically to COX2 / Cyclooxygenase 2. A band was observed at 75 kDa in wild-type A549 cell lysates with no signal observed at this size in PTGS2 knockout cell line ab280802 (knockout cell lysate ab283825). To generate this image, wild-type and PTGS2 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 fluorescent western blot (TBS-based) blocking solution 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation (ab179800).

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

PTGS2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human PTGS2 (COX2 / Cyclooxygenase 2) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-ptgs2-cox2-cyclooxygenase-2-knockout-a549-cell-line-ab280802'>ab280802</a>)

Lane 3:

U-87 MG cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Predicted band size: 69 kDa

false

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Compared with ab179800, ab283574 has higher sensitivity, we recommend ab283574 as an alternative for testing COX2 in western blot.

ab181602 was used as a loading control at a 1/1000000 dilution.

Blocking and dilution buffer : 5% NFDM/TBST.

This data was developed using the same antibody clone in a different buffer formulation (ab179800).

Lanes 1 - 3:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

Lanes 4 - 6:

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

Lanes 1 and 3:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 2 and 4:

PTGS2 knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 6:

A549 (Human lung carcinoma epithelial cell) 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 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 75 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] - BSA and Azide free (AB227528)

Blocking buffer and concentration : 5% NFDM/TBST  Diluting buffer and concentration : 5% NFDM/TBST COX2 is expressed at a low level in Raw264.7, mouse retina, hippocampus, heart, kidney etc. (PMID : 22015457, PMID : 26001832, PMID : 23045674, PMID : 33737575). This data was developed using the same antibody clone in a different buffer formulation (ab179800). The WB is using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-COX2 / Cyclooxygenase 2 antibody [EPR12012] (<a href='/en-us/products/primary-antibodies/cox2-cyclooxygenase-2-antibody-epr12012-ab179800'>ab179800</a>) at 1/1000 dilution

Lane 1:

B16-F10 (Mouse skin melanoma) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

Mouse retina tissue lysate at 20 µg

Lane 4:

Mouse hippocampus tissue lysate at 20 µg

Lane 5:

Mouse heart tissue lysate at 20 µg

Lane 6:

Mouse kidney tissue 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 1/20000 dilution

Predicted band size: 69 kDa

Observed band size: 72 kDa

false

Exposure time: 60s