A total of 20 μg of IPAs protein were separated on a Hoefer Mini VE system (GE) (Thermofisher), using a precast 8%–25% PhastGel. After, SDS/PAGE proteins were transferred to a nitrocellulose filter (Amersham). Membranes were then probed with rabbit anti-phospho-p38 (Thr180/Tyr182) (Cell Signaling); rabbit anti-p38 (Cell Signaling), rabbit anti-Procaspase 3 (Santa Cruz) and rabbit anti-caspase 3 (Santa Cruz); rabbit anti-nNOS (Cell Signaling); and rabbit anti-α/β-tubulin (Cell Signaling). Immunostaining was detected using horseradish peroxidase-conjugated anti-rabbit IgG (GE). Immunoblots were revealed by the ECL prime (Amersham) and quantitated by densitometry using ImageJ software (NIH).
Rabbit anti phospho p38 thr180 tyr182
Manufactured by Cell Signaling Technology
Sourced in United States
Rabbit anti-phospho-p38 (Thr180/Tyr182) is a primary antibody that recognizes the phosphorylated form of p38 MAPK at the Thr180 and Tyr182 residues. This antibody is designed for the detection and quantification of activated p38 MAPK by techniques such as Western blotting.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti p38, by Cell Signaling Technology (2 mentions)
Rabbit anti caspase 3, by Santa Cruz Biotechnology (1 mentions)
Nitrocellulose filter, by Cytiva (1 mentions)
Rabbit anti nnos, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated anti rabbit igg, by GE Healthcare (1 mentions)
Ecl prime, by Cytiva (1 mentions)
Rabbit anti procaspase 3, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti α β tubulin, by Cell Signaling Technology (1 mentions)
Mouse anti α tubulin, by Abcam (1 mentions)
Avidin biotin complex kit, by Vector Laboratories (1 mentions)
Alexa fluor 680, by Thermo Fisher Scientific (1 mentions)
Rat anti mouse f4 80, by Bio-Rad (1 mentions)
Goat anti collagen 4, by Southern Biotech (1 mentions)
Rabbit anti phospho jnk thr183 tyr185, by Cell Signaling Technology (1 mentions)
Rat anti mouse ly6g, by Abcam (1 mentions)
Rabbit anti α sma, by Abcam (1 mentions)
Pdvf membrane, by Merck Group (1 mentions)
Irdye infrared dye labeled secondary antibodies, by LI COR (1 mentions)
3 protocols using rabbit anti phospho p38 thr180 tyr182
1
Protein Expression Analysis by Western Blot
2
Immunohistochemical Analysis of Inflammation
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Antibodies used in this study were rat anti-mouse F4/80 (Bio-Rad, Gladesville, NSW, Australia); rat anti-mouse Ly6G, rabbit anti-α-SMA, and mouse anti-α-tubulin (Abcam, Melbourne, Victoria, Australia); goat anti-collagen IV (Southern Biotechnology, Birmingham, AL, USA); rabbit anti-phospho-p38 (Thr180/Tyr182) and rabbit anti-phospho-JNK (Thr183/Tyr185) (Cell Signaling, San Diego, CA, USA). Biotinylated goat anti-rabbit IgG was from Zymed-Invitrogen (Carlsbad, CA, USA). The avidin–biotin complex (ABC) kit was from Vector Labs (Burlingame, CA, USA). Alexa Fluor 680 or donkey anti-mouse IRDye 800 secondary antibodies were from Molecular Probes (Thermo Fisher Scientific, Waltham, MA, USA).
3
Quantitative Protein Expression Analysis
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Protein extracts from cells were obtained using RIPA 1X lysis buffer (#9806 Cell signaling, MA, were sonicated for 10 s and centrifuged at 9000g. Proteins were quantified with the Micro BCA assay kit following the manufacturer's instructions (Pierce, IL, USA). Extracts were subjected to SDS-PAGE electrophoresis in 9% polyacrylamide gels, transferred to PDVF membranes (Millipore, CA, USA), and probed with primary antibodies: rabbit anti-phospho-p38 (Thr180/Tyr182) (#9211S, Cell Signaling, USA), rabbit anti-p38 (#9212, Cell Signaling, USA) and mouse anti-b-actin (#21001901, AbLab, BC, CA). Then, primary antibodies were detected with IRDye® Infrared Dye labeled secondary antibodies (LI-COR). All immunoreactions were visualized by with an Odyssey® imaging system. Western blot densitometry quantification was done using Fiji (ImageJ) software. Protein levels were normalized with the levels of the loading control.
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