As described before [17 (link)], aliquots of 40 μg of protein from each group was separated by 13% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto a 0.22-μm polyvinylidene difluoride (PVDF) membrane (Millipore, Bedford, MA, USA). After blocking with 5% nonfat milk in TBS + 0.05% Tween™ 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C. Densitometric levels of CXCL1 signals were quantified and expressed as their ratio to GAPDH.
Rabbit anti phospho nf κb p65 ser536 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody is a primary antibody that detects the phosphorylated form of the NF-κB p65 subunit at serine 536. It is intended for use in western blotting and immunohistochemistry applications to study the activation of the NF-κB signaling pathway.
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Lab products found in correlation
2 protocols using rabbit anti phospho nf κb p65 ser536 antibody
1
Western Blot Analysis of CXCL1 and NF-κB Signaling
2
Immunofluorescence Staining of Phospho-NFκB p65
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The cells were seeded on round coverslips (Ø 10 mm) and after treatment they were washed with PBS, fixed with 4% paraformaldehyde for 10 min, permeabilized with PBS containing 0.1% Triton X-100 and incubated with a blocking buffer (4% horse serum in PBS). The cells were then subjected to immunofluorescence staining with a rabbit anti-Phospho-NFκB p65 (Ser536) antibody (Cell Signaling, 3033) (diluted 1:100 in a blocking buffer) overnight at 4 °C. After that, the cells were washed with PBS and incubated with Alexa 568-labeled anti-rabbit secondary antibody (diluted 1:1000 in a blocking buffer) (Invitrogen, A-11011) at room temperature for 1 h. Slides were mounted using ProLong Gold antifade with a DAPI reagent (Invitrogen, P36931). Digital images were acquired using a Carl Zeiss Axio Vert.A1 inverted microscope (ZEISS, Oberkochen, Germany) with an Ec Plan Neofluar 10× 0.3 Ph1 objective.
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