Anti-Phospho-HSL (S853) LIPE Rabbit Monoclonal Antibody

Western blot analysis of P-LIPE using anti-P-LIPE antibody (P06762-1).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat skeletal muscle tissue lysates,
Lane 2: mouse skeletal muscle tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-P-LIPE antigen affinity purified monoclonal antibody (P06762-1) at 1:1000 overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for P-LIPE at approximately 83 kDa. The expected band size for P-LIPE is at 117 kDa.

IHC analysis of P-LIPE using anti-P-LIPE antibody (P06762-1).
P-LIPE was detected in a paraffin-embedded section of human testis tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-P-LIPE Antibody (P06762-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Levels of proteins related to lipolysis after 6 weeks of 3-OBA and ET intervention. A , G Western blot (WB) analysis of P-HSL and HSL. B , H WB analysis of P-ATGL and ATGL. ( C , I ) WB analysis of CPT1b. D , J Ratio of P-HSL/HSL. E , K P-ATGL/ATGL ratio. F , L Fold protein of CPT1b. Three bands per group of rats are used. Relative levels were standardized to GAPDH. Measurement data were presented as the mean ± SD. The significance of the difference between the two groups C and H was calculated with the independent samples t test, and the four groups (S, 3, SH, and 3H) were analyzed with the two-way ANOVA. * p < 0.05 and ** p < 0.01. The number of biological repeats ( n = 10)
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