Anti-IL-12B antibody [EPR5739]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-IL-12B antibody [EPR5739] (AB133752)

Immunocytochemistry/ Immunofluorescence analysis of MOLT-4 (Human lymphoblastic leukemia T lymphoblast) cells labeling IL-12B with Purified ab133752 at 1 : 100 dilution (3.3 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were not counterstained. Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-IL-12B antibody [EPR5739] (AB133752)

Intracellular Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling IL-12B (red) with ab133752 at a 1/2500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• WB

Lab

Western blot - Anti-IL-12B antibody [EPR5739] (AB133752)

The recombinant protein contains Human IL-12B (Ile23-Ser328) and Human IL-12A (Arg23-Ser219) connected by disulfide bond to be heterodimer. Under reducing conditions, the observed bands will be at 40 KD for IL-12B and 29 KD for IL-12A.

Lanes 1 - 3:

Western blot - Anti-IL-12A antibody [EPR5736] (<a href='/en-us/products/primary-antibodies/il-12a-antibody-epr5736-ab133751'>ab133751</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-IL-12B antibody [EPR5739] (ab133752) at 1/1000 dilution

All lanes:

Human IL-12 recombinant protein at 15 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: 25 kDa,37 kDa

Observed band size: 29 kDa,40 kDa

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• WB

Unknown

Western blot - Anti-IL-12B antibody [EPR5739] (AB133752)

This image was generated using the unpurified version of the product.

All lanes:

Western blot - Anti-IL-12B antibody [EPR5739] (ab133752) at 1/1000 dilution

All lanes:

Human IL-12B recombinant protein at 0.01 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 37 kDa

Observed band size: 40 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-IL-12B antibody [EPR5739] (AB133752)

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.