Rabbit anti p38 antibody

Manufactured by Cell Signaling Technology

Sourced in United States

The Rabbit anti-P38 antibody is a primary antibody that specifically binds to the p38 MAPK protein. It can be used for the detection and analysis of p38 MAPK in various applications, such as Western blotting, immunoprecipitation, and immunohistochemistry.

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5 protocols using rabbit anti p38 antibody

Western Blot Analysis of Signaling Proteins

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Subconfluent cells were washed with PBS, and lysed with RIPA lysis buffer (150 mmol/L NaCl, 50 mmol/L Tris–HCl (pH 7.4), 1 % Triton X-100, 1 % sodium deoxycholate, 0.1 % SDS) with 1 mM sodium orthovanadate, 1 mM PMSF, and 1 % cocktail of protease inhibitors (Sigma). The lysates were clarified by centrifugation at 12,000g for 20 min at 4 °C and separated by SDS-PAGE followed by transfer onto nitrocellulose membranes. The following antibodies were used: rabbit anti-P38 antibody, rabbit anti-P-P38 antibody, rabbit anti-P-Akt antibody, mouse anti-HSP27 antibody and rabbit anti-P-HSP27 antibody (Cell Signaling, Danvers, MA, USA), rabbit anti-Akt antibody (Bioworld, Louis Park, USA), rabbit anti-GAPDH antibody (Santa Cruz, CA, USA). Protein bands were detected by incubating with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz) and visualized with ECL reagent (Millipore, Billerica, MA, USA). Digital images of immunoblots were obtained with a Chemidoc XRS and analyzed using the image analysis program Quantity One (Bio-Rad, Hercules, CA, USA).

Deng W., Zhang Y., Gu L., Cui J., Duan B., Wang Y, & Du J. (2016). Heat shock protein 27 downstream of P38-PI3K/Akt signaling antagonizes melatonin-induced apoptosis of SGC-7901 gastric cancer cells. Cancer Cell International, 16, 5.

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Investigating SKOV-3 Ovarian Cancer Cells

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Human ovarian adenocarcinoma cells (SKOV-3) were purchased from American Type Culture Collection (Manassas, VA, USA). RES was purchased from MilliporeSigma (Burlington, MA, USA). MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was purchased from MilliporeSigma. Guava^® Cell Cycle Reagent was purchased from Luminex Corporate (Austin, TX, USA). Annexin V was purchased from ImmunoTools (Friesoythe, Niedersachsen, Germany). Propidium iodide was purchased from MilliporeSigma. Rabbit anti-phospho p38 antibody, rabbit anti p38 antibody, rabbit anti-phospho ERK1/2 antibody, mouse anti-ERK1/2 antibody, rabbit anti-phospho JNK antibody, rabbit anti-JNK antibody, rabbit anti-phospho AKT (Ser473) antibody, mouse anti-AKT antibody, rabbit anti-cyclin A2 antibody, rabbit anti-cyclin B1 antibody, rabbit anti-cyclin E1 antibody, rabbit anti-caspase-9 antibody, rabbit anti-caspase-3 antibody, rabbit anti-PARP antibody, mouse anti-β-actin antibody, and DAPI (4, 6-diamidino-2-phenylindole, dihydrochloride) were purchased from Cell Signaling Technology (Danvers, MA, USA). Goat anti-mouse IgG-IRDye^®800CW and goat anti-rabbit IgG-IRDye^®680RT were purchased from Li-COR Biosciences (Lincoln, NE, USA). Goat anti-rabbit conjugated with Alexa488 and Goat anti-mouse conjugated with Alexa594 were purchased from Thermo Fisher Scientific (Waltham, MA, USA).

Hankittichai P., Thaklaewphan P., Wikan N., Ruttanapattanakul J., Potikanond S., Smith D.R, & Nimlamool W. (2023). Resveratrol Enhances Cytotoxic Effects of Cisplatin by Inducing Cell Cycle Arrest and Apoptosis in Ovarian Adenocarcinoma SKOV-3 Cells through Activating the p38 MAPK and Suppressing AKT. Pharmaceuticals, 16(5), 755.

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Ano1 and TRPV3 Inhibitor Assay Protocol

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T16Ainh-A01 (T16A, Calbiochem) and Ani9 (Sigma-Aldrich) were used as an Ano1 inhibitor. Dyclonine (MedChemExpress) was used as a TRPV3 inhibitor. The following antibodies were used: rabbit anti-ANO1 antibody (Abcam, ab53213, 1:5 for Western blotting), (Abcam, ab53212, 1:100 for immunoprecipitation), rabbit anti-phospho-ERK (extracellular signal-related kinase) antibody (Cell Signaling Technology, #4370, 1:1000), rabbit anti-phospho-p38 antibody (Cell Signaling Technology, #4511, 1:1000), rabbit anti-phospho-JNK (c-Jun N-terminal Kinase) antibody (Cell Signaling Technology, #4668, 1:1000), rabbit anti-ERK antibody (Cell Signaling Technology, #4695, 1:1000), rabbit anti-p38 antibody (Cell Signaling Technology, #8690, 1:1000), rabbit anti-JNK antibody (Cell Signaling Technology, #9252, 1:1000), mouse anti-β-actin antibody (Abcam, ab6276, 1:2500), rabbit anti-TRPV3 antibody (Cell Signaling Technology, #3484, 1:1000 for Western blotting; 1:50 for immunoprecipitation), anti-rabbit-HRP antibody (Cell Signaling Technology, #7074, 1:2000) and anti-mouse-HRP antibody (Cell Signaling Technology, #7076, 1:2000).

Yamanoi Y., Lei J., Takayama Y., Hosogi S., Marunaka Y, & Tominaga M. (2023). TRPV3-ANO1 interaction positively regulates wound healing in keratinocytes. Communications Biology, 6, 88.

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Histological and Immunohistochemical Analysis of Paraffin-Embedded Tissues

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Paraffin-embedded tissues were cut in the thickness of 5 μm, mounted onto glass slides, baked for 1 h at 60°C, deparaffinized in Xylene (Cat. No. 10023418, SCR, China), and sequentially rehydrated in 99, 95, and 70% ethanol (Cat. No. 10009218, SCR, China). Tissue slides were counterstained with Haematoxylin (Mayer's) (Cat. No. MB9897, Meilunbio, China) and Eosin (Cat. No. MA0164, Meilunbio, China) before dehydration with 95 and 99% ethanol, and were mounted with neutral balsam (Cat. No. 1004160, SCR, China). Stained tissues were analyzed under a light microscope (Leica DM IL LED). For immunohistochemical staining, paraffin-embedded tissue sections were stained with a rabbit anti-PDGFRβ antibody (Cat. No. ab32570, Abcam; 1:200); a mouse anti-CD163 antibody (Cat. No. ab156769, Abcam; 1:200); a rabbit anti-αSMA antibody (Cat. No. ab32575, Abcam; 1:200); a rabbit anti-phospho-p38 antibody (Cat. No. 4631S, Cell Signaling; 1:100), and a rabbit anti-p38 antibody (Cat. No. 9212S, Cell Signaling; 1:100). A ready to use HP IHC detection kit (Cat. No. abs957, Absin, China) was used for visualization. Captured images were further analyzed using Adobe Photoshop CS software.

Yang L., Sun X., Ye Y., Lu Y., Zuo J., Liu W., Elcock A, & Zhu S. (2019). p38α Mitogen-Activated Protein Kinase Is a Druggable Target in Pancreatic Adenocarcinoma. Frontiers in Oncology, 9, 1294.

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Molecular Profiling of Ocular Angiogenesis

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Fresh isolated eye tissues and cells were lysed, quantified, blotted and developed as previously described. 18 The following primary antibodies were used for the purpose of this experiment: goat anti-mouse endocan antibody (0.1 lg/mL; R&D Systems); rabbit anti-matrix metalloproteinase (MMP)-2 antibody (1:1000; Cell Signaling Technology); MMP-9 antibody (1:1000; Cell Signaling Technology); rabbit anti-PIGF antibody (1:1000; Abcam, Cambridge, UK); goat anti-VEGF antibody (1:1000; Abcam); rabbit anti-VEGFR1 antibody (1:1000; Abcam); rabbit anti-VEGFR2 antibody (Cell Signaling Technology); rabbit anti-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-phospho (p)-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-extracellular signal-regulated kinase (ERK) antibody (1:1000; Cell Signaling Technology); rabbit anti-p-ERK (1:1000; Cell Signaling Technology); rabbit anti-c-Jun Nterminal kinases (JNK) antibody (1:1000; Cell Signaling Technology) and rabbit anti-p-JNK antibody (1:1000; Cell Signaling

Su T., Zhong Y., Demetriades A.M., Shen J., Sui A., Yao Y., Gao Y., Zhu Y., Shen X, & Xie B. (2018). Endocan Blockade Suppresses Experimental Ocular Neovascularization in Mice. Investigative ophthalmology & visual science, 59(2).

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