NF-kB Signaling Pathway Antibody Sampler Panel (ab228529)

Overview

  • Product name

    NF-kB Signaling Pathway Antibody Sampler Panel
  • Product overview

    ab228529, NF-kB Signaling Pathway Antibody Panel, is a collection of 1 anti-rabbit (HRP) secondary antibody and 5 recombinant rabbit monoclonal antibodies against IKB alpha (p S36), IKB alpha, NFkB p105, NF-kB p65 (p S536), and NF-kB p65 provided in 10 µl trial sizes.

  • Notes

    Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.

    Directly conjugated versions of our antibodies are available and ready to use for multicolor flow cytometry or immunocytochemistry analysis. Carrier-free formulations are also available for easy conjugation to labels of your choice. Please refer to the ‘Associated products’ section below.

Properties

Images

  • ICC/IF image of unpurified ab32518 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32518, 1/1000 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Predicted band: 105 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: NFkB p105 / p50 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32360 observed at 120, 50 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32360 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE. ab32360 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of colon sections from mice, staining NF-kB p65 with ab32536.

    Antigen retrieval was performed by microwave heating in citrate buffer, pH 6. Sections were incubated overnight with primary antibody (1/250) and staining was detected using ab80437 EXPOSE Rabbit specific HRP/DAB detection IHC kit.

  • Lane 1:

    Primary - ab133462, 1/10000 dilution

    Sample - HeLa cell lysate, 10 µg

    Secondary - HRP labeled goat anti-rabbit IgG, 1/2000 dilution

    Lane 2:

    Primary - ab133462, 1/10000 dilution

    Sample - HeLa cell lysate treated with FBS and Calyculin A, 10 µg

    Secondary - HRP labeled goat anti-rabbit IgG, 1/2000 dilution

    Predicted band: 35 kDa

    Actual band: 40 kDa

References

ab228529 has not yet been referenced specifically in any publications.

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