Anti-IL-10 antibody [EPR1114]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-10 antibody [EPR1114] (AB133575)

Intracellular Flow Cytometry analysis of permeabilized HeLa cells labelling IL-10 using unpurified ab133575 at a 1/100 dilution (red) or a control rabbit IgG antibody (green).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-10 antibody [EPR1114] (AB133575)

Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab133575 at a dilution of 1 in 300 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IL-10 antibody [EPR1114] (AB133575)

Immunofluorescence staining of Molt-4 cells with purified ab133575 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab133575 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

• ICC/IF

AbReview45709****

Immunocytochemistry/ Immunofluorescence - Anti-IL-10 antibody [EPR1114] (AB133575)

Unpurified ab133575 staining IL-10 in human peripheral blood mononuclear cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Saponin in PBS and blocked with 4% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG polyclonal (1/100) was used as the secondary antibody.

This image is courtesy of an anonymous Abreview

• Flow Cyt (Intra)

AbReview45706****

Flow Cytometry (Intracellular) - Anti-IL-10 antibody [EPR1114] (AB133575)

Intracellular flow cytometric analysis ofhuman PBMCs (peripheral blood mononuclear cells)treated with PHA labeling IL-10 with ab133575 at 1/100 dilution (green), followed by Goat anti-rabbit Alexa Fluorr^® 488.

1 (dashed line) = isotype control, 2 (black line) = PBMCs untreated, 3 (green line) = PBMCs treated with PHA.

This image is courtesy of an anonymous Abreview.

• WB

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Western blot - Anti-IL-10 antibody [EPR1114] (AB133575)

All lanes:

Western blot - Anti-IL-10 antibody [EPR1114] (ab133575) at 1/1000 dilution

All lanes:

Recombinant IL-10 protein at 0.005 µg

Secondary

All lanes:

Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 20 kDa

Observed band size: 18 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-IL-10 antibody [EPR1114] (AB133575)

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.