Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

Immunocytochemistry/Immunofluorescence analysis of HaCaT (Human keratinocyte cell line) labelling Topoisomerase I with purified ab131166 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131166).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

Clone EPR5376(2) (ab240047) has been successfully conjugated by Abcam. This image was generated using Anti-Topoisomerase I antibody [EPR5376(2)] (Alexa Fluor® 647). Please refer to ab223422 for protocol details.

ab223422 staining Topoisomerase I in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab223422 at 1/50 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in MCF7 cells fixed with 100% methanol (5 min).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

Clone EPR5376(2) (ab240047) has been successfully conjugated by Abcam. This image was generated using Anti-Topoisomerase I antibody [EPR5376(2)] (Alexa Fluor® 488). Please refer to ab223421 for protocol details.

ab223421 staining Topoisomerase I in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab223421 at 1/100 dilution (shown in Green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in MCF7 cells fixed with 100% methanol (5 min).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

Immunofluorescent analysis of MCF7 cells labelling Topoisomerase I with ab131166 at 1/250 dilution.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131166).

• WB

Lab

Western blot - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

Blocking and diluting buffer : 5%NFDM/TBST. We recommend to use 1%SDS Hot lysis prepare method. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131166).

All lanes:

Western blot - Anti-Topoisomerase I antibody [EPR5376(2)] (<a href='/ko/products/primary-antibodies/topoisomerase-i-antibody-epr53762-ab131166'>ab131166</a>) at 1/10000 dilution

Lane 1:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 15 µg

Lane 3:

SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 15 µg

Lane 4:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/2000 dilution

Predicted band size: 91 kDa

Observed band size: 91 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Topoisomerase I antibody [EPR5376(2)] - BSA and Azide free (AB240047)

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.