Ab131435

Manufactured by Abcam

Sourced in United States, China, United Kingdom

Ab131435 is a polyclonal antibody that recognizes the protein target. It is intended for use in laboratory research applications.

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Lab products found in correlation

4 protocols using ab131435

Western Blot Analysis of Signaling Proteins

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Cell lysates were separated by SDS-PAGE, followed by wet transfer onto 0.45 µm PVDF membranes, which were then blocked with 5% BSA in PBST solution for 30 min. Specific primary antibody incubation was conducted overnight at 4 °C. The antibodies used were as follows: anti-phospho-Y397-Fak (ab81298, Abcam, USA), anti-FAK (ab131435, Abcam, USA), anti-Tyr416-phosphor-c-Src (#2010s, CST), anti-c-Src (#2108s, CST), anti-Ser473-phospho-Akt (#4060s, CST), anti-Akt (#9272, CST, Danvers, MA), anti-FASN (#14979-1-AP, Proteintech, Rosemont, USA), anti-β-actin (#sc-81178, Santa Cruz, USA). Secondary antibody was incubated for 1 h at RT. Immunodetection was accomplished using enhanced chemiluminescence. The band intensity was determined with ImageJ software. All the original western blot pictures were provided in the “Supplemental Material original western blots”.

Du Q., Liu P., Zhang C., Liu T., Wang W., Shang C., Wu J., Liao Y., Chen Y., Huang J., Tan H., Zhao Y., Xia M., Liu J, & Yao S. (2022). FASN promotes lymph node metastasis in cervical cancer via cholesterol reprogramming and lymphangiogenesis. Cell Death & Disease, 13(5), 488.

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Protein Expression Analysis in Tumor Samples

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Proteins extracted from tumor tissue and cells were separated by Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and transferred to Polyvinylidene Fluoride (PVDF) membranes (Sigma Aldrich). Following blockade with bovine serum albumin (Sigma Aldrich) and incubation with primary antibodies against DSG3 (1:2000, ab218380, Abcam), Src (ab47405) and p-Src (1:2500, p-Tyr418, ab4816 Abcam), FAK (ab131435) and p-FAK (1:3000, p-Tyr397, ab81298, Abcam), AKT (ab235958) and p-AKT (1:3500, p-Ser 473, ab81283, Abcam), and β-actin (1:4000, ab8227, Abcam), the membrane was then incubated with HRP-conjugated secondary antibody (1:5000, ab205718, Abcam), and the signals were detected with chemiluminescence system (Tanon, Shanghai, China).

Abula Y., Su Y., Tuniyazi D, & Yi C. (2021). Desmoglein 3 contributes to tumorigenicity of pancreatic ductal adenocarcinoma through activating Src–FAK signaling. Animal Cells and Systems, 25(3), 195-202.

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Western Blot Analysis of Cell Signaling

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Western blot analyses were performed using an anti-alpha V antibody (ab124968, and ab112487 Abcam, Cambridge, MA), phospho-FAK antibody (ab24781, Abcam, Cambridge, MA) FAK antibody (ab131435, Abcam, Cambridge, MA), TLR2 antibody (ab213676, Abcam, Cambridge, MA), TLR4 antibody (SC-293072, Santa Cruz Biotecnology, Santa Cruz, CA), or MyD88 antibody (ab2064, Abcam, Cambridge, MA) followed by a horseradish peroxidase-conjugated anti-mouse antibody (SC-2005, Santa Cruz Biotechnology). Blots were then developed with the ECL-plus detection system (Thermo Fisher Scientific, Waltham, MA). To evaluate the samples for equal protein loading, membranes were stripped and re-probed with an anti-β-actin antibody (SC-1615, Santa Cruz Biotechnology).

Kamarajan P., Ateia I., Shin J.M., Fenno J.C., Le C., Zhan L., Chang A., Darveau R, & Kapila Y.L. (2020). Periodontal pathogens promote cancer aggressivity via TLR/MyD88 triggered activation of Integrin/FAK signaling that is therapeutically reversible by a probiotic bacteriocin. PLoS Pathogens, 16(10), e1008881.

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Investigating FAK and Src Signaling

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1-CP-U, generously provided by Ms. Qizhi Ning, was diluted to the required concentrations in conditional medium and then stored at 4°C according to the reported procedure [19]. Matrigel was purchased from Corning, USA (356234). The antibody to FAK (ab131435) and p-FAK Tyr397 (ab81298) were from Abcam, UK. p-Src Tyr416 antibody (6943) and paxillin antibody (12065) were purchased from Cell Signaling Technology, MA, USA. Anti-β-actin, goat anti-mouse and anti-rabbit secondary antibodies were from Boster Biological Technology (CA, USA).

Peng J., Yang X., Li X., Gao H., Liu N, & Guo X. (2019). 1-calcium phosphate-uracil inhibits intraperitoneal metastasis by suppressing FAK in epithelial ovarian cancer. Cell Cycle, 18(16), 1925-1937.

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