Deparaffinized liver sections were used for immunostaining of TNF-α and Caspase-3 as described previously [45 (link)]. Caspase-3 immunostaining was done using rabbit polyclonal anti-cleaved Caspase-3 antibody diluted 1:100 (BioCare Medical, Cat. CP229C, Concord, CA, USA). TNF-α immunostaining was done using a polyclonal rabbit anti-TNFα antibody diluted 1: 400 (GeneTex, Cat# GTX110520). Twenty sections were examined in each group. To determine the Caspase-3 and TNF-α immunoreactivity, the DAB-stained cytoplasmic option in the IHC profiler Image-J plugin was used as described previously [46 (link)] in the captured photomicrographs. Finally, the area% of Caspase-3 and TNF-α expressions was measured using Image J software (freely available public domain image processing software) as described previously [47 (link)].
Rabbit anti tnfα antibody
Manufactured by GeneTex
Rabbit anti-TNFα antibody is a primary antibody that recognizes and binds to the tumor necrosis factor alpha (TNFα) protein. It can be used in various research applications to detect and study the expression and function of TNFα.
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2 protocols using rabbit anti tnfα antibody
1
Immunostaining of Liver Tissue
2
Western Blot Analysis of Brain Proteins
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Western blot analysis was performed as previously described [38 (link)]. For details, see SI Methods . Briefly, mice were sacrificed by rapid decapitation under deep anesthesia. The brain tissues were dissected in ice-cold 1× PBS buffer, then the samples were homogenized and centrifuged. Total protein concentration was determined using an Enhanced BCA Protein Assay Kit (Beyotime). The following antibodies were used: rabbit anti-PV antibody (1:4000, Abcam), rabbit anti-Arg1 antibody (1:5000, Abcam), rabbit anti-β-tubulin antibody (1:5000, Cell Signaling Technology), rabbit anti-IL1β antibody (1:2000, Abcam), rabbit anti-IL6 antibody (1:3000, GeneTex), rabbit anti-TNFα antibody (1:3000, GeneTex), and goat anti-rabbit IgG (H + L) HRP (1:8000, Abways). Protein bands were visualized using a chemiluminescence system (ChemiDocTM XRS+, BioRad). The protein expressions were semi-quantitatively analyzed with ImageJ software.
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