Western Blot Analysis of Neuroinflammatory Markers

Separated skin and L3–L5 dorsal horn specimens from all rats were processed using a western blotting protocol described previously [9 (link)], using antibodies against the following proteins: COX-2 (#12282, 1:1000 dilution; Cell Signaling), iNOS (ab15323, 1:1000; Abcam), nNOS (ab76067, 1:1000; Abcam), Akt/protein kinase B (#4685, 1:1000; Cell Signaling), p-Akt (#4060, 1:1000; Cell Signaling), B-cell lymphoma 2 (ab59348, Bcl-2; 1:1000; Abcam), Bcl-2-associated X protein (50599-2-Ig, Bax; 1:1000; Proteintech Group, Chicago, IL, USA), β-actin (A5441, 1:20000; Sigma-Aldrich), LC3B (1:1000; Cell Signaling) and Beclin 1 (#2775, 1:1000; Cell Signaling). The membranes were visualized by Bio-Rad ChemiDoc MP and band intensity was quantitated by Quantity One 1-D Analysis software (Bio-Rad Laboratories Inc., Hercules, CA, USA).

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Lin C.H., Wu S.H., Lee S.S., Lin Y.N., Kuo Y.R., Chai C.Y, & Huang S.H. (2017). Autologous Adipose-Derived Stem Cells Reduce Burn-Induced Neuropathic Pain in a Rat Model. International Journal of Molecular Sciences, 19(1), 34.

Publication 2017

COX-2 ab15323 ab76067 Akt/protein kinase B p-Akt β-actin Beclin 1 Bio-Rad ChemiDoc MP Quantity One 1-D Analysis software

Actin Antibodies B cell lymphoma Bcl 2 Bcl 2 associated x protein Beclin 1 Cox 2 Dilution Dorsal horn Inos Membranes Nnos Protein kinase b Proteins Rats Skin

Corresponding Organization : Kaohsiung Medical University Chung-Ho Memorial Hospital

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Variable analysis

independent variables

Separated skin and L3–L5 dorsal horn specimens from all rats

dependent variables

Protein levels of COX-2, iNOS, nNOS, Akt/protein kinase B, p-Akt, Bcl-2, Bax, LC3B, and Beclin 1

control variables

β-actin protein level

controls

Positive control: None mentioned
Negative control: None mentioned

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