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Phospho smad1 ser463 465 smad5 ser463 465 smad9 ser465 467

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The Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad9 (Ser465/467) is an antibody that detects endogenous levels of Smad1, Smad5, and Smad9 proteins when phosphorylated at the specified serine residues.

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Lab products found in correlation

Phospho nf κb p65 ser536 93h1 rabbit mab, by Cell Signaling Technology (2 mentions) Smad1 d59d7 xp rabbit mab, by Cell Signaling Technology (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Nf κb p65 d14e12 xp rabbit mab, by Cell Signaling Technology (1 mentions) Nitrotyrosine, by Abcam (1 mentions) Phosphorylated enos, by Cell Signaling Technology (1 mentions) Pkg1α, by Enzo Life Sciences (1 mentions) Caveolin 1, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Abcam (1 mentions) Xp rabbit mab, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

SMAD5 (18 citations) Smad9 (2 citations)

3 protocols using phospho smad1 ser463 465 smad5 ser463 465 smad9 ser465 467

1

Western Blot Analysis of Smad and NF-κB Signaling

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Cells were lysed with RIPA Lysis and Extraction Buffer (Thermo Scientific cat.# PI89900) buffer with protease/phosphatase inhibitor cocktail. The lysate was centrifuged at 12,000×g at 4°C for 15 min, boiled with Laemmli buffer for 7 min at 95°C, and transferred to PVDF membranes (Millipore). After blocking, membranes were incubated with primary antibody at 4°C overnight followed by incubation with corresponding secondary HRP-linked antibody. The following antibodies were used for western blotting: Smad1 (D59D7) XP Rabbit mAb (Cell Signaling Technology cat.# 6944), Phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad9 (Ser465/467) (Cell Signaling Technology cat.# 13,820P), NF-κB p65 (D14E12) XP Rabbit mAb (Cell Signaling Technology cat.# 8242), Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Cell Signaling Technology cat.# 3033), Anti-GAPDH antibody EPR16884 Loading Control (Abcam cat.# ab181603).
Xiong L., Zhevlakova I., West X.Z., Gao D., Murtazina R., Horak A., Brown J.M., Molokotina I., Podrez E.A, & Byzova T.V. (2024). TLR2 regulates hair follicle cycle and regeneration via BMP signaling. eLife, 12, RP89335.
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2

Western Blot Analysis of Lung Proteins

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Protein samples from whole lung tissues or cultured cells were subjected to Western blot analysis with antibodies against Cavin-2 (Proteintech), Caveolin-1 (Santa Cruz Biotechnology), phosphorylated eNOS (Cell Signaling), eNOS (Cell Signaling), nitrotyrosine (Abcam), PKG-1α (Enzo), phospho-Smad1 (Ser463/465)/Smad5 (Ser463/465)/Smad9 (Ser465/467) (Cell Signaling), Smad1 (Cell Signaling), phospho-MLC2 at Ser19 (Cell Signaling), MLC (Cell signaling), and Gapdh (Abcam). Signal intensities were determined using Image J software (National Institutes of Health, Bethesda, MD). Uncropped data of Western blots are provided in Figs. S7–S9.
Kasahara T., Ogata T., Nakanishi N., Tomita S., Higuchi Y., Maruyama N., Hamaoka T, & Matoba S. (2023). Cavin-2 loss exacerbates hypoxia-induced pulmonary hypertension with excessive eNOS phosphorylation and protein nitration. Heliyon, 9(6), e17193.
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3

Western Blot Analysis of Smad and NF-κB Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed with RIPA Lysis and Extraction Buffer (ThermoScientific™cat.#PI89900) buffer with protease/phosphatase inhibitor cocktail. The lysate was centrifuged at 12000g at 4°C for 15 min, boiled with Laemmli buffer for 7 min at 95°C, and transferred to PVDF membranes (Millipore). After blocking, membranes were incubated with primary antibody at 4°C overnight followed by incubation with corresponding secondary HRP-linked antibody. The following antibodies were used for western blotting: Smad1 (D59D7) XP® Rabbit mAb (Cell Signaling Technology cat#6944), Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad9 (Ser465/467) (Cell Signaling Technology cat# CLS8160-24EA), NF-κB p65 (D14E12) XP® Rabbit mAb (Cell Signaling Technology cat#8242), Phospho-NF-κB p65 (Ser536) (93H1) Rabbit mAb (Cell Signaling Technology cat#3033), Anti-GAPDH antibody EPR16884 Loading Control (Abcam cat#ab181603).
Xiong L., Zhevlakova I., West X.Z., Gao D., Murtazina R., Horak A., Brown J.M., Molokotina I., Podrez E.A, & Byzova T.V. (2023). TLR2 Regulates Hair Follicle Cycle and Regeneration via BMP Signaling. bioRxiv.
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