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Total nf κb p65

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The Total NF-κB p65 assay is a quantitative enzyme-linked immunosorbent assay (ELISA) used to measure the total amount of the NF-κB p65 subunit in cell and tissue samples. NF-κB p65 is a transcription factor involved in regulating cellular responses to various stimuli, including inflammation, immunity, and cell survival.

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Lab products found in correlation

Phospho nf κb p65, by Santa Cruz Biotechnology (1 mentions) Phospho stat3, by Santa Cruz Biotechnology (1 mentions) Phospho akt, by Santa Cruz Biotechnology (1 mentions) Phospho stat5, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Santa Cruz Biotechnology (1 mentions) Phospho syk, by Cell Signaling Technology (1 mentions) Total erk1 2, by Cell Signaling Technology (1 mentions) Total iκbα, by Cell Signaling Technology (1 mentions) Total ikkβ, by Cell Signaling Technology (1 mentions) Total foxo1, by Cell Signaling Technology (1 mentions) Phospho foxo1 foxo3a, by Cell Signaling Technology (1 mentions) C myc, by Cell Signaling Technology (1 mentions) Phospho s6, by Cell Signaling Technology (1 mentions) Nf κb2 p100 p52, by Cell Signaling Technology (1 mentions) Phospho iκbα, by Cell Signaling Technology (1 mentions) Phospho akt, by Cell Signaling Technology (1 mentions) Phospho nf κb p65, by Cell Signaling Technology (1 mentions) Phospho erk1 2 thr202 tyr204, by Cell Signaling Technology (1 mentions)

2 protocols using total nf κb p65

1

Western Blot Analysis of Signaling Proteins

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Tissues or cells were mechanically homogenized in ice-cold lysis buffer and centrifuged for 10 minutes at 4 °C and 14,000 g for tissues or 8,000 g for cells. The resulting supernatant fraction was separated by SDS-PAGE and immunoblotted with antibodies against phospho-AKT (S473; 1:1000), total AKT (1:1000), total JAK2 (1:1000), tubulin (1:1000), phospho-STAT3 (Y705, 1:2000), total STAT3 (1:1000), phospho-STAT5 (Y694, 1:1000), total STAT5 (1:1000), phospho-NF-κB p65 (Y694, 1:1000), total NF-κB p65 (1:1000), phospho-JNK (T183/Y185, 1:500, Santa Cruz Biotechnology) or total JNK1 (1:500, Santa Cruz Biotechnology) or GAPDH (1:5000). Antibodies were purchased from Cell Signaling Technology unless otherwise stated. Protein band intensity was quantified by ImageJ software.
Desai H.R., Sivasubramaniyam T., Revelo X.S., Schroer S.A., Luk C.T., Rikkala P.R., Metherel A.H., Dodington D.W., Park Y.J., Kim M.J., Rapps J.A., Besla R., Robbins C.S., Wagner K.U., Bazinet R.P., Winer D.A, & Woo M. (2017). Macrophage JAK2 deficiency protects against high-fat diet-induced inflammation. Scientific Reports, 7, 7653.
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2

Signaling Pathway Analysis by Western Blot

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Whole cell lysates were prepared by direct lysing and boiling samples in Laemmli buffer supplemented with β-mercaptoethanol. The samples were subjected to Western blotting and membranes were incubated with one of the following antibodies: Phospho-IκBα (Ser32; clone: 14D4, Cell Signaling Technology), total-IκBα (clone: L35A5, Cell Signaling Technology), Phospho-IKKα/IKKβ (Ser176/Ser177; clone: C84E11, Cell Signaling Technology), total-IKKβ (clone: D30C6, Cell Signaling Technology), Actin (clone: C4/Actin, BD Biosciences), Phospho-Erk1/2 (Thr202/Tyr204; clone: D13.14.4E, Cell Signaling Technology), total-Erk1/2 (clone: 137F5, Cell Signaling Technology), Phospho-NF-κB p65 (Ser536; clone 93H1, Cell Signaling Technology), total- NF-κB-P65 (rabbit polyclonal, Santa Cruz Biotechnology), c-Myc (clone: D84C12, Cell Signaling Technology), Phospho-Syk (Tyr352; rabbit polyclonal, Cell Signaling Technology), total-Syk (clone: 5F5, BioLegend), Phospho-Akt (Ser473; clone: D9E, Cell Signaling Technology), Phospho-S6 (Ser235/236; clone:D57.2.2E, Cell Signaling Technology), Phospho-FoxO1/FoxO3a (Thr24/Thr32; rabbit polyclonal, Cell Signaling Technology), total-Foxo1 (clone: C29H4, Cell Signaling Technology), NF-κB2 p100/p52 (rabbit polyclonal, Cell Signaling Technology). The signals on the membranes were quantitated by ImageJ software. Loading was normalized by Actin blotting.
Luo W., Weisel F, & Shlomchik M.J. (2018). B cell receptor and CD40 Signaling are Rewired for Synergistic induction of the c-Myc transcription factor in Germinal Center B cells. Immunity, 48(2), 313-326.e5.
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