Protein extracts were prepared with cytoplasmic lysis buffer (25mM Tris-HCl [pH7.4], 40mM KCl, and 1% Triton X-100) supplemented with 1 mg/ml aprotinin and 1 mg/ml leupeptin (all obtained from Sigma). Twenty micrograms of total protein was run on 10%−15% SDS-polyacrylamide gels, and Western blotting was performed exactly as described (Theurl et al., 2006 (link)) with a mouse anti-human TfR1 antibody (1:1000; Invitrogen), rabbit anti-human Ferritin (1:500; Sigma), rabbit anti-Fpn (1:400; self designed; Eurogentic), rabbit anti-HO-1 (1:1000; Enzo), rabbit anti-iNOS (1:1000; BD), rabbit anti-NFκB p65 or rabbit anti-phosphor NFκB p65 (both 1:1000; Cell Signalling). Blotting with either rabbit anti-TBP (1:1000; Cell Signalling) or rabbit anti-Actin antibody (1:1000; Sigma-Aldrich) was performed as a loading control.
Rabbit anti phosphor nfκb p65
Manufactured by Cell Signaling Technology
Rabbit anti-phosphor NFκB p65 is a primary antibody product from Cell Signaling Technology. It is designed to detect phosphorylated forms of the NF-κB p65 subunit.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti actin antibody, by Merck Group (1 mentions)
Rabbit anti nf κb p65, by Cell Signaling Technology (1 mentions)
Rabbit anti tbp, by Cell Signaling Technology (1 mentions)
Mouse anti human tfr1 antibody, by Thermo Fisher Scientific (1 mentions)
Rabbit anti human ferritin, by Merck Group (1 mentions)
Rabbit anti inos, by BD (1 mentions)
Rabbit anti ho 1, by Enzo Life Sciences (1 mentions)
Aprotinin, by Merck Group (1 mentions)
Leupeptin, by Merck Group (1 mentions)
Confocal microscope, by Zeiss (1 mentions)
Alexa fluor 488 conjugated goat anti rabbit igg, by Abcam (1 mentions)
Rabbit anti tgf β1, by Proteintech (1 mentions)
Rabbit anti pparγ, by Abcam (1 mentions)
2 protocols using rabbit anti phosphor nfκb p65
1
Western Blot Analysis of Iron Metabolism Proteins
2
Immunofluorescence Analysis of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Formaldehyde‐fixed lung sections (5 μm) were dewaxed in xylene and rehydrated in ethanol/water. After antigen repair solution, 0.05% Triton X‐100 permeabilization and blocking with 10% goat serum, the sections were incubated with primary antibodies, including rabbit anti‐PPAR‐γ (Abcam), rabbit anti–phosphor‐Stat6 (Cell Signaling Technology), rabbit anti‐TGF‐β1 (ProteinTech), rabbit anti‐phosphor‐Smad3 (Cell Signaling Technology), rabbit anti‐phosphor‐NFκB P65 (Cell Signaling Technology) and rabbit/mouse anti‐CD68 (Abcam) at 4℃ overnight, respectively. Sections were washed three times and incubated with Alexa Fluor 488‐conjugated goat anti‐rabbit IgG (Abcam), Alexa Fluor 647‐conjugated donkey anti‐mouse IgG (Abcam) and CoralLite 594‐conjugated goat anti‐rabbit IgG (ProteinTech) at 37℃ for 1 h, and then labelled with DAPI for 5minutes. Immunofluorescence was measured by a confocal microscope (Carl Zeiss).
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