Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free (AB248471)

This data was developed using ab133319, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of Paraffin Embedded Human stomach tissue labelling COX1/Cyclooxygenase 1 with ab133319 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free (AB248471)

This data was developed using ab133319, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] (<a href='/en-us/products/primary-antibodies/cox1-cyclooxygenase-1-antibody-epr5867-ab133319'>ab133319</a>) at 1/1000 dilution

Lane 1:

C2C12 cell lysate at 10 µg

Lane 2:

A431 cell lysate at 10 µg

Lane 3:

NIH/3T3 cell lysate at 10 µg

Lane 4:

Neuro 2a cell lysate at 10 µg

Lane 5:

L6 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 68 kDa

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• WB

Lab

Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free (AB248471)

This data was developed using ab133319, the same antibody clone in a different buffer formulation. False colour image of Western blot : Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] 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, ab133319 was shown to bind specifically to COX1 / Cyclooxygenase 1. A band was observed at 70 kDa in wild-type A431 cell lysates with no signal observed at this size in PTGS1 knockout cell line ab270477 (knockout cell lysate ab270500). To generate this image, wild-type and PTGS1 knockout A431 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.

Lanes 1 - 4:

Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] (<a href='/en-us/products/primary-antibodies/cox1-cyclooxygenase-1-antibody-epr5867-ab133319'>ab133319</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free (ab248471) at 1/1000 dilution

Lane 1:

Wild-type A431 cell lysate at 20 µg

Lane 2:

Western blot - Human PTGS1 knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-ptgs1-knockout-a-431-cell-line-ab270477'>ab270477</a>) at 20 µg

Lane 2:

Western blot - Human PTGS1 knockout A-431 cell lysate (ab270500) at 20 µg

Lane 2:

PTGS1 knockout A431 cell lysate at 20 µg

Lane 3:

C2C12 cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 70 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-COX1 / Cyclooxygenase 1 antibody [EPR5867] - BSA and Azide free (AB248471)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.