The following reagents and materials were used: anti-GAPDH (Calbiochem), Anti-OXSR1, anti-NONO (Abcam), anti-Prohibitin-1 (cell signaling), anti-dimethylated histone H3 (Lys9) (Upstate), anti-p38 MAP kinase, anti-phospho p38 MAP kinase (Thr180/Tyr 182), anti-PDK1, anti-phospho PDK1 (S241), anti-PKC-Pan, anti-phospho PKC-pan (T514), anti-pAkt (Ser473), anti-Akt, anti-pERK1/2 (Thr202/Tyr204), anti-ERK1/2, anti-STAT3, anti-phospho STAT3 (Y705), anti-cJUN, anti-phospho cJUN (S73), anti-NF-κB p65, anti-NF-κB phospho-p65 (S536), anti-PLCgamma, anti-phospho PLCgamma (Y783), anti-SAPK/JNK, and anti-phospho SAPK/JNK (T183/Y185) (Cell Signaling). Electrophoresis reagents were purchased from Bio-rad and trypsin from Promega.
Anti phospho pkc pan
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-phospho PKC-pan product is a laboratory reagent that detects the phosphorylated form of the pan-protein kinase C (PKC) family. It can be used to identify and quantify the activation state of PKC proteins in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti akt, by Cell Signaling Technology (2 mentions)
Anti phospho pdk1, by Cell Signaling Technology (2 mentions)
Anti pdk1, by Cell Signaling Technology (2 mentions)
Trypsin, by Promega (2 mentions)
Anti fibronectin, by Abcam (2 mentions)
Anti pkc, by Abcam (2 mentions)
Anti sglt2, by Abcam (2 mentions)
Anti tgf β, by Cell Signaling Technology (2 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti stat3, by Cell Signaling Technology (1 mentions)
Anti pkc pan, by Cell Signaling Technology (1 mentions)
Anti sapk jnk, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Anti erk1 2, by Cell Signaling Technology (1 mentions)
Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Anti phospho sapk jnk t183 y185, by Cell Signaling Technology (1 mentions)
Anti p erk1 2, by Cell Signaling Technology (1 mentions)
Anti c jun, by Cell Signaling Technology (1 mentions)
Anti pka c alpha, by Cell Signaling Technology (1 mentions)
Anti phospho mek1 2, by Cell Signaling Technology (1 mentions)
4 protocols using anti phospho pkc pan
1
Comprehensive Protein Signaling Assay
2
Characterizing Kinase Signaling Pathways
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The following reagents and materials were used. Anti-p38 MAPK (ref. 9212), anti-phospho-p38 MAPK (T180/Y182) (ref. 9211), anti-Akt (ref. 4685), anti-phospho-Akt (S473) (ref. 4060), anti-PKA C-alpha (ref. 4782), anti-phospho-PKA C (T197) (ref. 5661), anti-SEK1 (ref. 9152), anti-phospho-SEK1 (S257/T261) (Ref. 9156), anti-MEK1/2 (ref. 9126), anti-phospho-MEK1/2 (S217/221) (ref. 9154), anti-phospho GSK3 α/β (S21) (ref. 9331), anti-GSK3 α/β (ref. 5676), anti-Phb1 (ref. 2426), and anti-Phb2 (ref. 14085) anti-PDK1 (ref. 3062), anti-phospho-PDK1 (S241) (ref. 3061), anti-phospho-PKC pan (T514) (ref. 9379), anti-MEK1/2 (ref. 9126), anti-phospho-MEK1/2 (S217/221) (ref. 9154), anti-phospho CAMKII (Y86) (ref.12716) and anti-CAMKII (ref.11945) were purchased from Cell Signaling Technology. Anti-PKC-pan was from Sigma Aldrich (ref. SAB4502356). Electrophoresis reagents were purchased from Bio-rad and trypsin from Promega.
3
Antibody Sourcing Protocol
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Anti-NOX4, anti-PKC, anti-SGLT2, anti-fibronectin antibodies were purchased from Abcam (Cambridge, MA, USA). Anti-pan phospho-PKC, anti-TGF-β antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Horseradish peroxidase (HRP)-conjugated donkey anti-rabbit immunoglobulin G (IgG) secondary antibody was obtained from Amersham Biosciences (GE Healthcare Bio-Sciences Corp., Piscataway, NJ, USA). Anti-β-actin HRP-conjugated antibody was obtained from Santa Cruz (Dallas, TX, USA). All other chemicals and reagents were purchased from standard suppliers unless otherwise mentioned.
4
Protein Expression Analysis of Mesangial Cells
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After 72 h of incubation with or without canagliflozin in the presence of 5.5 mM or 25 mM glucose, MCs were collected and sonicated in 400 µl of solubilizing solution containing 9 M urea, 2% Triton X-100, and 1% dithiothreitol. Then, 100 µl of 10% lithium dodecyl sulfate was added to the solubilized membrane fraction and sonicated again. Subsequently, proteins were separated discontinuously on 7.5% sodium dodecyl sulfate polyacrylamide gels and transferred to PVDF membranes (Bio-Rad, Hercules, CA, USA). After blocking nonspecific binding, the membranes were incubated overnight at 4 °C with rabbit anti-NOX4 (1:1000; Abcam), anti-PKC (1:250; Abcam), anti-pan phospho-PKC (1:1000; Cell Signaling Technology), anti-SGLT2 (1:500; Abcam), anti-TGF-β (1:500; Cell Signaling Technology), or anti-fibronectin (1:5000; Abcam) antibodies, followed by HRP-conjugated donkey anti-rabbit IgG antibody (1:10,000; Amersham) as a secondary antibody. The membranes were also incubated with goat anti-β-actin HRP-conjugated antibody (1:5000; Santa Cruz) to ensure equal protein loading of the lanes. We used the ECL Plus system (Amersham) to detect antibody binding.
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