RIPA Buffer (Tris–HCl 25 mM pH 7.4, NaCl 150 mM, 1% Triton X-100, Sodium Deoxycholate 1%, SDS 0.1%, EDTA 2 mM) containing a protease/phosphatase inhibitor mixture (Roche) was utlized to lyse cells. After electrophoresis with SDS-PAGE, the separated proteins from the gel (50 μg) were transferred on nitrocellulose (NC) membranes, followed by incubation with primary antibodies. Following incubation with appropriate HRP-labeled secondary antibodies, ECL HRP substrate (Beyotime) was employed to detect signals. The primary antibodies used were as follow: α-SMA (55135-1-AP, Proteintech), Collagen I (14695-1-AP, Proteintech), Timp-1 (CSB-PA023560YA01HU, CUSABIO, Wuhan, China), Vimentin (10366-1-AP, Proteintech), Erk (16443-1-AP, Proteintech), p-Erk (sc-81492, Santa Cruz Biotechnology, Santa Cruz, CA, USA), p38 (14064-1-AP, Proteintech), p-p38 (AF4001, Affinity, Changzhou, China), JNK (AF6319, Affinity), p-JNK (AF3318, Affinity), NF-κB (AF5006, Affinity), p-NF-κB (AF2006, Affinity).
P erk sc 81492
Manufactured by Santa Cruz Biotechnology
Sourced in United States
P-Erk (sc-81492) is a lab equipment product offered by Santa Cruz Biotechnology. It is an antibody that detects phosphorylated forms of Erk1 and Erk2 proteins.
Automatically generated - may contain errors
Lab products found in correlation
P jnk, by Affinity Biosciences (1 mentions)
Vimentin, by Proteintech (1 mentions)
α sma 55135 1 ap, by Proteintech (1 mentions)
Erk 16443 1 ap, by Proteintech (1 mentions)
Af3318, by Affinity Biosciences (1 mentions)
Protease phosphatase inhibitor mixture, by Roche (1 mentions)
Collagen 1 14695 1 ap, by Proteintech (1 mentions)
Af5006, by Affinity Biosciences (1 mentions)
Nf κb, by Affinity Biosciences (1 mentions)
Sc 47778, by Santa Cruz Biotechnology (1 mentions)
Zdr 5307, by ZSGB-BIO (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Immobilon western chemiluminescence hrp substrate, by Merck Group (1 mentions)
Ab32360, by Abcam (1 mentions)
Jnk sc 7345, by Santa Cruz Biotechnology (1 mentions)
P jnk sc 6254, by Santa Cruz Biotechnology (1 mentions)
Xar film, by Kodak (1 mentions)
Erk sc 514302, by Santa Cruz Biotechnology (1 mentions)
Nf κb p65, by Cell Signaling Technology (1 mentions)
4 protocols using p erk sc 81492
1
Western Blot Analysis of Fibrosis Markers
2
Western Blot Analysis of Renal Protein
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Proteins from renal tissue or cell cultures were extracted and quantified using the bicinchoninic acid method. Then, equivalent concentrations of protein (40 μg/lane) were separated on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis gels and then transferred to a nitrocellulose membrane. Primary antibodies against p-ERK (sc-81492), ERK2 (sc-514302), p-JNK (sc-293136), JNK2 (sc-271133), c-fos (sc-52), p-p38 (sc-7973), and β-actin (sc-47778) were purchased from Santa Cruz (dilution 1:1,000), and p38 (#8690), p-CREB (#9198), and CREB (#9197) were purchased from Cell Signaling Technology (dilution 1:1,000). β-actin was used as a loading control to ensure equal loading. Subsequently, membranes were washed twice with PBS and then incubated with goat anti-rabbit or goat anti-mouse HRP-conjugated immunoglobulin G secondary antibody (1:2,000; ZDR-5306, ZDR-5307, ZSGB-BIO, Beijing, China) at room temperature for 1 h. Specific bands were then visualized using Immobilon Western Chemiluminescence HRP substrate (Merck Millipore, Darmstadt, Germany). Optical densities were detected using Quantity One software (Bio-Rad, Her-cules, CA, USA).
3
Western Blot Profiling of Signaling Proteins
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Proteins were extracted from harvested cells, and their concentration was determined by the bicinchoninic acid assay (pierce). Protein samples (30 µg) were resolved by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. The following antibodies were used: anti‐mouse RANKL (FL‐317; Santa Cruz), nuclear factor‐κB (NF‐κB) P50 (ab32360; Abcam), NF‐κB P65 (#3033; Cell signaling), JNK (sc‐7345; Santa Cruz), p‐JNK (sc‐6254; Santa Cruz), ERK (sc‐514302; Santa Cruz), p‐ERK (sc‐81492; Santa Cruz). The results were visualized with Kodak autoradiography film (Kodak XAR film).
4
Extraction and Analysis of Gelidium crinale
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Fresh Gelidium crinale were collected from about 1-m depth of Naozhou Island Sea, Zhanjiang City, Guangdong Province, in summer 2020. Algae were identified through the morphological characters of the herbarium and the appraisal scheme of the Prof. Zhang C (Guangdong Ocean University).
The standards (fucose, galactose, glucuronic acid, rhamnose, arabinose, ribose, xylose, glucose, and aminogalactose) were provided by Sigma-Aldrich (Sigma Chemicals, St. Louis, MO, USA). The bicinchoninic acid (BCA) assay kit and all cell culture chemicals were provided by Thermo Fisher Scientific, Inc. (Waltham, MA, USA). LPS, 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse polyclonal antibodies, including p65 (sc-8008), p-p65 (sc-136548), IκBα (sc-1643), p-IκBα (sc-8404), p-JNK (sc-6254), JNK (sc-7345), p-p38 (sc-166182), and p-ERK (sc-81492); rabbit polyclonal antibodies (p38, (sc-535); ERK (sc-94)); and secondary antibodies, such as goat anti-rabbit IgG-HRP (sc-2004), and goat anti-mouse IgG-HRP (sc-2005) were provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA).
The standards (fucose, galactose, glucuronic acid, rhamnose, arabinose, ribose, xylose, glucose, and aminogalactose) were provided by Sigma-Aldrich (Sigma Chemicals, St. Louis, MO, USA). The bicinchoninic acid (BCA) assay kit and all cell culture chemicals were provided by Thermo Fisher Scientific, Inc. (Waltham, MA, USA). LPS, 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse polyclonal antibodies, including p65 (sc-8008), p-p65 (sc-136548), IκBα (sc-1643), p-IκBα (sc-8404), p-JNK (sc-6254), JNK (sc-7345), p-p38 (sc-166182), and p-ERK (sc-81492); rabbit polyclonal antibodies (p38, (sc-535); ERK (sc-94)); and secondary antibodies, such as goat anti-rabbit IgG-HRP (sc-2004), and goat anti-mouse IgG-HRP (sc-2005) were provided by Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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