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2 protocols using anti rabbit igg hrp linked antibody

1

Western Blot Analysis of Protein Expression

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RIPA lysate buffer containing 1 mM PMSF was added to each sample to collect the total protein. The total protein concentration of each sample was determined by the BCA method and adjusted all samples to the same concentration. The protein samples were mixed with a 5x loading buffer and denatured at 95°C. The proteins were separated by SDS-PAGE (8% or 15%) and electrophoretically transferred onto polyvinylidene fluoride membranes. The membranes were probed with rabbit anti-TrkB antibody (1 : 1000; Cell Signaling Technology; Cat. No. #4603), rabbit anti-CREB antibody (1 : 1000; Cell Signaling Technology; Cat. No. #9197S), rabbit anti-p-CREB antibody (1 : 1000; Cell Signaling Technology; Cat. No. #9198S), rabbit anti-β-tubulin antibody (1 : 1000; Cell Signaling Technology; Cat. No. #2128), rabbit anti-synaptophysin polyclonal antibody (1:1000; Proteintech; Cat. No. #17785-1-AP), and rabbit anti-GAPDH antibody (1 : 1000; Servicebio; Cat. No. #GB11002) overnight at 4°C and then incubated with Anti-rabbit IgG HRP-Linked Antibody (1 : 3000; Servicebio; Cat. No. #GB23303) at 37°C for 1.5 h. Detection was performed using a ChemiDoc XRS+ (Bio-Rad, USA) image analysis system.
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2

Antioxidant and Anti-inflammatory Effects of XYW

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The XYW (Tai Ji, China, batch number 1707029) and fluoxetine hydrochloride (FLX) (Patheon, France, 7686 A) were dissolved in pure water solution preparation (SOD, batch number 20180,309), malondialdehyde (MDA, batch number 20180,313) (GSH, batch number 20180,621), and GSH/glutathione disulfide (GSH/GSSG, batch number 20180,726) levels were assessed using commercially available kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Nitric oxide (NO) assay kit (Biyuntian, 0,72117,171,110, China). Rat interleukin 6 (IL6) ELISA Kit (MULTI SCIENCES, A30680231), rat interleukin 1β (IL1B) ELISA Kit (ExCell Bio, 21H183), rabbit anti-NTRK2 antibody (Cell Signaling Technology; Cat. No. #4603), rabbit anti-AKT1 antibody (Cell Signaling Technology; Cat. No. #4691S), rabbit anti-p-AKT1 antibody (Cell Signaling Technology; Cat. No. #4060S), rabbit anti-HMOX1 antibody (Cell Signaling Technology; Cat. No. #S2206S), rabbit anti-PIK3CA antibody (Servicebio; Cat. No. #LS190932), rabbit anti-GAPDH antibody (Servicebio; Cat. No. #GB11002), rabbit anti-SOD1 antibody (Novus Biologicals; Cat. No. #03253462C2B), rabbit anti-NFE2L2 antibody (Invitrogen; Cat. No. #TK2668681A), rabbit anti-brain-derived neurotrophic factor (BDNF) antibody (Abcam; Cat. No. #ab108319) and anti-rabbit IgG HRP-linked antibody (Servicebio; Cat. No. #GB23303).
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