IKKβ Antibody (H-4): sc-8014

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Serum-starved HPAEpiCs were incubated with TNF-α at 37°C for the various time points. At the end of treatment, the cells were harvested and centrifuged at 45000 × g at 4°C for 1 h to prepare the whole cell lysate, as previously described (Chi et al., 2012). The denatured samples were analyzed by 10% SDS-PAGE gels and transferred to nitrocellulose membrane. The Western blot was performed by incubation membrane with an anti-cPLA2, anti-p47phox, anti-Gαs, anti-TRAF2, anti-ASK1, anti-TNFR1, anti-NIK, anti-IKKα, anti-IKKβ, anti-p65, anti-phospho-serine or anti-GAPDH (1:1000) antibody for 24 h, and then incubated with an anti-mouse horseradish peroxidase antibody (1:2000) for 1 h. ECL reagents and UVP BioSpectrum 500 Imaging System (Upland, CA, USA) were used to detect and capture the immunoreactive bands. The image densitometry of each immunoreactive band was analyzed and quantified by the UN-SCAN-IT gel software (Orem, UT, USA).

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Manufacturer protocol

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