Total protein from PC12 cells or the hippocampal tissues was extracted and analyzed using Western blots. Prepared samples were separated by 10% or 12% SDS-PAGE and transferred to Polyvinylidene Fluoride (PVDF) membranes (Millpore, Bedford, MA, USA). The membranes were incubated with anti-GAPDH (Abcam, Cambridge, UK), anti-thr181-phosphorylated-tau, anti-thr205-phosphorylated-tau, anti-ser396-phosphorylated-tau, anti-total tau, anti-JNK, anti-phospho JNK, anti-ERK1/2, anti-phospho ERK1/2, anti-phospho p38, anti-p38, anti-NF-κB, anti-phospho NF-κB (CST), anti-IL-6, and anti-TNF-α (Proteintech, Wuhan, China) antibodies. Immunoreactive bands were detected with the appropriate horseradish peroxidase-conjugated secondary antibodies and immunological complexes were visualized by enhanced chemiluminescence reagents (Pierce, Rockford, IL, USA).
Anti phospho erk1 2
Manufactured by Proteintech
Sourced in China, United States
Anti-phospho ERK1/2 is a primary antibody that binds to the phosphorylated forms of extracellular signal-regulated kinase 1 and 2 (ERK1/2). ERK1/2 are important signaling molecules that play a role in cellular processes such as proliferation, differentiation, and survival.
Automatically generated - may contain errors
Lab products found in correlation
Anti erk1 2, by Proteintech (3 mentions)
Anti nf κb, by Proteintech (1 mentions)
Anti il 6, by Proteintech (1 mentions)
Anti phospho p38, by Proteintech (1 mentions)
Anti phospho jnk, by Proteintech (1 mentions)
Anti p38, by Proteintech (1 mentions)
Anti tnf α, by Proteintech (1 mentions)
Anti gapdh, by Abcam (1 mentions)
Anti jnk, by Proteintech (1 mentions)
Anti p16, by Proteintech (1 mentions)
Anti phospho p38 mapk, by Proteintech (1 mentions)
Anti phospho jnk, by Cell Signaling Technology (1 mentions)
Anti jnk, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Proteintech (1 mentions)
Anti p53, by Proteintech (1 mentions)
Anti p38 mapk, by Proteintech (1 mentions)
Cell lysis solution, by Cell Signaling Technology (1 mentions)
Anti cd31, by Proteintech (1 mentions)
Ecl system, by Beyotime (1 mentions)
Anti snail, by Proteintech (1 mentions)
4 protocols using anti phospho erk1 2
1
Western Blot Analysis of Neuroinflammatory Markers
2
Protein Expression Analysis via SDS-PAGE
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SDS polyacrylamide gel electrophoresis tests were performed as the previous study [25 (link)]. Protein expression levels were detected by ECL luminescence imaging. The primary antibodies used in this experiment included anti-PRDM9(PA541161;Invitrogen); anti-FBLN5(Catalog # 3095-FB; R&D system); anti-p16 (Cat No. 10883-1-AP;proteintech), anti-p53(Cat No. 60283-2-Ig; proteintech), anti-phospho-p38 MAPK(Cat No. 28796-1-AP ;Proteintech), anti-p38MAPK(Cat No. 14064-1-AP ;Proteintech), anti-phospho-Erk1/2(Cat No: 80031-1-RR;Proteintech), anti-Erk1/2(CatNo. 11257-1-AP; Proteintech); anti-phospho-JNK (CatNo. 4668; Cell Signaling Technology), anti‐JNK (Cat No. 9258; Cell Signaling Technology) and anti-GAPDH (Cat No. 60004-1-1g; Proteintech)
3
Western Blot Analysis of HGEC Proteins
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Whole‐cell lysates were produced using a cell lysis solution from Cell Signalling Technology (Danvers, MA). The proteins were denatured by heating in loading buffer at 100°C for 10 min. Equal amounts of protein (50 μg) from all HGEC groups were separated using 8%–10% SDS‐PAGE and subsequently transferred onto PVDF membranes. These membranes were then blocked using a protein‐free rapid‐blocking solution (Beyotime Biotechnology, China) for 1 h. After the blocking step, the membranes underwent overnight incubation with designated primary antibodies at 4°C. The antibodies utilized in this investigation included anti‐β‐actin, anti‐HIC1, and anti‐Sirt7; anti‐SDC1, anti‐H3K18ac, anti‐CD31, anti‐αSMA, anti‐Snail, anti‐ERK1/2, and anti‐phospho‐ERK1/2 (ProteinTech, Wuhan, China). After undergoing five washes, the membranes were treated by secondary antibodies at ambient temperature for an hour, followed by another five washes with PBST. The protein signals were detected using an ECL system (Beyotime Institute of Biotechnology, Shanghai, China).
4
Western Blot Analysis of GC Cell Proteins
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In a radio-immunoprecipitation assay (RIPA) lysis buffer, the proteins were extracted from GC cells. Ten percents of polyacrylamide sodium dodecyl sulfate (SDS) gels (SDS-PAGE) were used to separate equivalent amounts of proteins. The proteins were then blotted onto membranes of polyvinylidene fluoride (Amersham, UK). After membranes were blocked at RT (37°C) in bovine serum albumin (5%) for 1 h, primary antibodies were used to culture the membranes overnight. After being cultured, they were placed in secondary antibodies conjugated with human resource planning (Perbio Science, Belgium), and incubated at RT (37°C) for 1 h. The system of enhanced chemiluminescence immunodetection (Immobilon, USA) was used to visualize immunoreactive proteins. The antibodies for Western blotting were as follows: anti-KIF22 (13403-1-AP; Proteintech Group, USA), anti-Phospho-MEK1/2 (CST: #9154; Proteintech Group), anti-MEK (CST: #8727; Cell Signaling Technology, USA), anti-Phospho-ERK1/2 (CST: #4370; Proteintech Group), anti-ERK (16443-1-AP; Proteintech Group), anti-Cyclin A2 (18202-1-AP; Proteintech Group), anti-Cyclin B1 (55004-1-AP; Proteintech Group), anti-Cyclin D1 (26939-1-AP; Proteintech Group), anti-P21 (10355-1-AP; Proteintech Group), and β-actin (20536-1-AP; Proteintech Group).
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