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Anti phospho src family tyr416

Manufactured by Cell Signaling Technology
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The Anti-phospho-Src family (Tyr416) is a research-use only antibody that detects the phosphorylated form of the Src family kinases at the Tyr416 site. It can be used for applications such as Western blotting, immunoprecipitation, and immunohistochemistry.

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Lab products found in correlation

Anti phospho akt ser473, by Cell Signaling Technology (3 mentions) Anti akt, by Cell Signaling Technology (2 mentions) Polyvinylidene fluoride membrane, by Merck Group (1 mentions) Anti phospho c fos, by Cell Signaling Technology (1 mentions) Anti p47phox, by Cell Signaling Technology (1 mentions) Anti phospho erk1 2, by Cell Signaling Technology (1 mentions) Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions) Anti phospho c jun, by Cell Signaling Technology (1 mentions) Pro preptm, by iNtRON Biotechnology (1 mentions) Anti erk, by Cell Signaling Technology (1 mentions) Anti rhoa sc 418, by Santa Cruz Biotechnology (1 mentions) Anti pyk2, by Merck Group (1 mentions) Anti p38α sc 535, by Santa Cruz Biotechnology (1 mentions) Immobilon p membrane, by Merck Group (1 mentions) Western lightning plus ecl, by PerkinElmer (1 mentions) Anti nfatc1, by BD (1 mentions) Anti src, by Cell Signaling Technology (1 mentions) Streptavidin hrp, by Cell Signaling Technology (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Apoptosis western blot cocktail, by Abcam (1 mentions)

4 protocols using anti phospho src family tyr416

1

Western Blot Analysis of Signaling Pathways

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Protein extraction solution PRO-PREPTM was obtained from iNtRON Biotechnology (Jungwon, Gyeonggi, Korea). Polyvinylidene fluoride membranes and western chemiluminescent HRPV substrate were obtained from Millipore Corporation (Billerica, MA, USA). The above reagents were used according to the manufacturer’s protocol. The following antibodies from Cell Signaling Technology (Danvers, MA, USA) were used: EGFR antibody, anti-phospho-Src family (Tyr416), anti-phospho-AKT (Ser473), anti-phospho-Erk1/2, anti-p47phox, anti-phospho-NF-κB p65, anti-phospho-c-Fos, anti-phospho-c-Jun, anti-p44/42 MAPK (ERK1/2), anti-NF-κB p65, and anti-β-actin monoclonal antibodies. The detailed western blot steps were described previously [13 (link)].
Li S., Nguyen T.T., Ung T.T., Sah D.K., Park S.Y., Lakshmanan V.K, & Jung Y.D. (2022). Piperine Attenuates Lithocholic Acid-Stimulated Interleukin-8 by Suppressing Src/EGFR and Reactive Oxygen Species in Human Colorectal Cancer Cells. Antioxidants, 11(3), 530.
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2

Western Blotting Analysis of Signaling Proteins

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Total cell extracts were obtained using lysis buffer containing 150mMTris-HCl (pH 6.8), 6% SDS, 30% glycerol, and 0.03% Bromophenol Blue; 10% 2-ME was added immediately before harvesting cells. Cell lysates were fractionated on 7.5% SDS-PAGE, transferred to Immobilon-P membranes (Millipore), and incubated with specific antibodies. Western Lightning plus-ECL (PerkinElmer) was used for detection. For immunoblotting, anti-NFATc1 (556602, 1:1000 dilution) was from BD Biosciences. Anti-Blimp1 (sc-47732, 1:1000 dilution), anti-Gα13 (sc-410, 1:500 dilution), anti-RhoA (sc-418), and anti-p38α (sc-535, 1:1000 dilution) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-pyk2 (06-559, 1:1000 dilution) was from EMD Millipore. Anti-phospho-Pyk2 (Try402) (#3291, 1:1000 dilution), anti-phospho-Akt (Ser473) (#9271, 1:1000 dilution), anti-phospho-Akt (Thr408) (#2965, 1:1000 dilution), anti-Akt (#9272, 1:1000 dilution), anti-phospho-ERK (Ser473) (#9101, 1:1000 dilution), anti-ERK (#9102, 1:1000 dilution), anti-phospho-Src family (Tyr416) (#2101, 1:1000 dilution) were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Anti- phospho-Rock2 (Ser1366, 1:500 dilution) is from GeneTex.
Nakano S., Inoue K., Xu C., Deng Z., Syrovatkina V., Vitone G., Zhao L., Huang X.Y, & Zhao B. (2019). G-protein Gα13 functions as a cytoskeletal and mitochondrial regulator to restrain osteoclast function. Scientific Reports, 9, 4236.
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3

Immunoblotting and Quantification Techniques

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Immunoprecipitation and immunoblotting experiments were performed as previously described 58 (link) using the following primary antibodies: anti-CD99 clone 12E7 (Dako, Inc.; #M3601); anti-CD99 clone 013 (Invitrogen, #180235 or MA5-12287); anti-CD99 antibody [EPR3097Y] (Abcam, #ab75858); anti-cyclophilin A (Abcam, #ab58144); anti-PKA-RIIα (Santa Cruz Biotechnology, #sc-908); anti-Src (Cell Signaling, #2123); anti-phospho-Src family (Tyr416) (Cell Signaling, #6943); streptavidin-HRP (Cell Signaling, #3999); anti-actin-HRP, (Santa Cruz Biotechnology, #sc-1615); anti-ROCK2 clone C20 (Santa Cruz Biotechnology, #sc-1851); anti-GAPDH (Santa Cruz Biotechnology, #sc-25778); and apoptosis western blot cocktail (Abcam, # ab136812). Quantitative analysis of protein bands was performed using ImageJ 1.48v software 59 (link).
Çelik H., Sciandra M., Flashner B., Gelmez E., Kayraklıoğlu N., Allegakoen D.V., Petro J.R., Conn E.J., Hour S., Han J., Oktay L., Tiwari P.B., Hayran M., Harris B.T., Manara M.C., Toretsky J.A., Scotlandi K, & Üren A. (2018). Clofarabine inhibits Ewing sarcoma growth through a novel molecular mechanism involving direct binding to CD99. Oncogene, 37(16), 2181-2196.
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4

Comprehensive Signaling Pathway Analysis

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Membranes were incubated with the following primary antibodies (1:1000 dilution, if not stated otherwise): anti-P-Tyr-1000 (Cell Signaling Technology cat# 8954), anti-Lck (Cell Signaling Technology cat# 2752), anti-Src L4A1 (Cell Signaling Technology cat# 2110), anti-phospho Lck (Tyr505) (Cell Signaling Technology cat# 2751), anti-phospho Src Family (Tyr416) (Cell Signaling Technology cat# 2101), anti-IGF1Rβ (Cell Signaling Technology cat# 3027), anti-phospho IGF1Rβ (Tyr1135) (Cell Signaling Technology cat# 3918), anti-phospho mTOR (Ser2448) (Cell Signaling Technology cat# 2971), anti-phospho p70 S6K (Thr421/Ser424) (Cell Signaling Technology cat# 9204), anti-AKT (Cell Signaling Technology cat# 9272), anti-phospho AKT (Ser473) (Cell Signaling Technology cat# 9271), anti-p44-42 MAPK (ERK1/2) (137F5) (Cell Signaling Technology cat# 4695), anti-phospho p44-42 MAPK (Thr202/Tyr204) (D13.14.4E) (Cell Signaling Technology cat# 4370), anti-cleaved caspase-3 (Asp175) (Cell Signaling Technology cat# 9661), and anti-β actin (Abcam, cat# ab6276, 1:10,000).
For protein bands staining, SDS-PAGE gels were stained using the Colloidal Blue Staining kit (Invitrogen cat# LC6025) according to the manufacturer protocol. Uncropped and unprocessed blots are provided in the Source Data file.
Cordo’ V., Meijer M.T., Hagelaar R., de Goeij-de Haas R.R., Poort V.M., Henneman A.A., Piersma S.R., Pham T.V., Oshima K., Ferrando A.A., Zaman G.J., Jimenez C.R, & Meijerink J.P. (2022). Phosphoproteomic profiling of T cell acute lymphoblastic leukemia reveals targetable kinases and combination treatment strategies. Nature Communications, 13, 1048.
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