Gadolinium chloride and Yoda1 were purchased from Tocris Bioscience (Bristol, UK). Ruthenium Red was purchased from Cayman Chemical (Ann Arbor, MI). GsMTx4 was purchased from both Tocris Bioscience and Alomone Labs (Jerusalem, Israel). Rabbit anti-phospho-Akt (Ser473) (#4060), rabbit anti-phospho-ERK1/2 (T202/Y204) (#9101), and rabbit anti-ERK1/2 (#9102) were obtained from Cell Signaling Technology (Danvers, MA). Goat anti-Akt (sc-1618) was from Santa Cruz Biotechnology (Dallas, Tx).
Rabbit anti phospho erk1 2 t202 y204
Manufactured by Cell Signaling Technology
Rabbit anti-phospho-ERK1/2 (T202/Y204) is a primary antibody that recognizes the phosphorylated forms of extracellular signal-regulated kinase 1 and 2 (ERK1/2) at threonine 202 and tyrosine 204. It is used to detect and quantify the activated, phosphorylated state of these important signaling proteins.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti akt, by Cell Signaling Technology (2 mentions)
Rabbit anti phospho akts473, by Cell Signaling Technology (2 mentions)
Yoda1, by Bio-Techne (1 mentions)
Ruthenium red, by Cayman Chemical (1 mentions)
Gsmtx4, by Bio-Techne (1 mentions)
Rabbit anti phospho akt ser473, by Cell Signaling Technology (1 mentions)
Rabbit anti erk1 2, by Cell Signaling Technology (1 mentions)
U0126, by Santa Cruz Biotechnology (1 mentions)
Millicell hanging cell culture insert, by Merck Group (1 mentions)
Goat anti actin 1 19 sc 1616, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti mmp 9, by Merck Group (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Tnf α, by ProSpec (1 mentions)
Ecl western blotting detection reagent, by GE Healthcare (1 mentions)
Anti rabbit igg hrp, by GE Healthcare (1 mentions)
Anti mouse igg hrp, by GE Healthcare (1 mentions)
Mouse anti egfr, by Cell Signaling Technology (1 mentions)
Rabbit anti phospho egfr y1068, by Cell Signaling Technology (1 mentions)
Mouse anti actin, by Merck Group (1 mentions)
Las 3000 imaging system, by Fujifilm (1 mentions)
4 protocols using rabbit anti phospho erk1 2 t202 y204
1
Pharmacological Modulation of Signaling Pathways
2
Quantifying Protein Signaling Cascades
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TNF-α was obtained from ProSpec-Tany TechnoGene Ltd. (cyt-223-b) and U0126 was purchased from Santa Cruz (CAS 109511–58-2). The antibodies used were: rabbit anti-phospho-ERK1/2 (T202/Y204, Cell Signaling), mouse anti-ERK1/2 (L34F12, Cell Signaling), rabbit anti-MMP-9 (AB19016, Merck Millipore), rabbit anti-MMP2 (VARP20016_T100, Aviva), rabbit anti-MMP14 (PA5–13,183, Thermo Scientific), rabbit anti-FAP-α (GTX102732, GeneTex), goat anti-DPPIV/CD26 (AF1180, R&D Systems), rabbit-anti-Cortactin (H-191) (sc-11,408, Santa Cruz Biotechnology, Inc.), goat anti-actin (I-19) (sc-1616, Santa Cruz Biotechnology, Inc.) and mouse anti-FL-2 (B6) (sc-28,320, Santa Cruz Biotechnology, Inc.). The cholera toxin subunit B-HRP conjugate was obtained from Molecular Probes (C-34,780). Millicell Hanging Cell Culture Inserts (8.0 μm) were purchased from Merck Millipore (PIEP15R48) and blotting membranes were purchased from Bio-Rad (162–0094). Immuno-detection was performed using HRP-conjugated donkey anti-rabbit (sc-2313), donkey anti-goat (sc-2020) and goat anti-mouse (sc-2005) antibodies purchased from Santa Cruz Biotechnology, Inc. and Pierce™ECL Western Blotting Substrate (32,106, Thermo Scientific).
3
Protein Extraction and Western Blot Analysis
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Whole‐cell extracts were prepared by lysing cells with RIPA buffer (50 mM Tris‐HCl [7.4], 150 mM NaCl, 1% Triton X‐100, 0.5% sodium deoxycholate, 0.1% SDS) containing 5 mM sodium vanadate, 5 mM NaF, and complete EDTA‐free protease inhibitor (Roche) for 30 minutes at 4°C. Insoluble materials were removed by centrifugation. Western blotting was carried out using standard methods. The following antibodies were used: rabbit anti‐phospho‐EGFR (Y1068) (1:3000; Cell Signaling #2237), mouse anti‐EGFR (1:3000; Cell Signaling #2239), rabbit anti‐phospho‐AKT (S473) (1:3000; Cell Signaling #9271), rabbit anti‐AKT (1:3000; Cell Signaling #9272), rabbit anti‐phospho‐ERK1/2 (T202/Y204) (1:3000; Cell Signaling #9101), rabbit anti‐ERK1/2 (1:10 000; Santa Cruz sc‐93), mouse anti‐actin (1:1000; Sigma C7207), anti‐mouse IgG‐HRP (1:10 000; GE Healthcare NA9310), and anti‐rabbit IgG‐HRP (1:10 000; GE Healthcare NA9340). Blots were developed using the ECL Western Blotting Detection Reagents (GE Healthcare RPN2109) and detected using the LAS‐3000 imaging system (Fujifilm).
4
Immunoblotting and Antibody Validation
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S1P was purchased from Cayman
Chemicals and JTE-013 from Sigma-Aldrich. Mouse anti-ERK 1/2 (1:1000,
clone 3A7), rabbit anti-phospho-ERK1/2 (T202/Y204) (1:1000, clone
D13.14.4E), rabbit anti-GFP (1:200, clone D5.1), rabbit anti-HA tag
(1:1000, clone C29F4), rabbit anti-p38 (1:1000, catalog no. 9212),
rabbit anti-phospho-p38 (T180/Y192) (1:1000, clone D3F9), rabbit anti-AKT
(1:1000, clone C67E7), and rabbit anti-phospho-AKT (S473) (1:1000,
clone D9E) antibodies were obtained from Cell Signaling. The mouse
anti-myc antibody (1:800, clone 9E10) was from Santa Cruz Biotechnology,
and the mouse anti-HA antibody (1:1000, clone 12CA5) was from Roche.
Rabbit anti-Flag (1:1000, clone F-7425), mouse anti-β-actin
(1:10000, clone A1978), and rabbit anti-β-catenin (1:1000, clone
C2206) antibodies were from Sigma-Aldrich. Goat anti-mouse Alexa 488
(1:1000), goat anti-rabbit Alexa 488 (1:1000), and goat anti-rabbit
Alexa 568 (1:1000) antibodies were purchased from Life Technologies.
Donkey anti-mouse IRDye800CW (1:20000) and donkey anti-mouse IRDye680LT
(1:20000) were bought from LI-COR Biosciences.
Chemicals and JTE-013 from Sigma-Aldrich. Mouse anti-ERK 1/2 (1:1000,
clone 3A7), rabbit anti-phospho-ERK1/2 (T202/Y204) (1:1000, clone
D13.14.4E), rabbit anti-GFP (1:200, clone D5.1), rabbit anti-HA tag
(1:1000, clone C29F4), rabbit anti-p38 (1:1000, catalog no. 9212),
rabbit anti-phospho-p38 (T180/Y192) (1:1000, clone D3F9), rabbit anti-AKT
(1:1000, clone C67E7), and rabbit anti-phospho-AKT (S473) (1:1000,
clone D9E) antibodies were obtained from Cell Signaling. The mouse
anti-myc antibody (1:800, clone 9E10) was from Santa Cruz Biotechnology,
and the mouse anti-HA antibody (1:1000, clone 12CA5) was from Roche.
Rabbit anti-Flag (1:1000, clone F-7425), mouse anti-β-actin
(1:10000, clone A1978), and rabbit anti-β-catenin (1:1000, clone
C2206) antibodies were from Sigma-Aldrich. Goat anti-mouse Alexa 488
(1:1000), goat anti-rabbit Alexa 488 (1:1000), and goat anti-rabbit
Alexa 568 (1:1000) antibodies were purchased from Life Technologies.
Donkey anti-mouse IRDye800CW (1:20000) and donkey anti-mouse IRDye680LT
(1:20000) were bought from LI-COR Biosciences.
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