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Rabbit anti smad1 antibody

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The Rabbit anti-Smad1 antibody is a primary antibody raised against the Smad1 protein. Smad1 is a key intracellular signal transducer and transcriptional modulator activated by bone morphogenetic protein (BMP) receptors. This antibody can be used to detect and analyze Smad1 expression and activation in various experimental systems.

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Lab products found in correlation

Startingblock buffer, by Thermo Fisher Scientific (1 mentions) Rabbit anti p65 antibody, by Cell Signaling Technology (1 mentions) Rabbit anti phosphor stat3, by Cell Signaling Technology (1 mentions) Mouse anti β actin, by Merck Group (1 mentions) Rabbit anti stat3, by Cell Signaling Technology (1 mentions)

Close spelling variants of this product

Rabbit anti-Smad1 Anti-Smad1 antibody Anti-Smad1

3 protocols using rabbit anti smad1 antibody

1

Shear Stress-Induced NF-κB and Smad1 Activation

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Cells were seeded on tissue culture plastic slides cut from 150 mm tissue culture dishes (Falcon), coated with 20 μg.ml−1 fibronectin. Confluent cells were subjected to steady laminar shear stress in a modified parallel plate flow chamber (Figure 1) in which the gasket was a silicon sheet of either 0.8 or 1.6 mm height (Grace Bio-Labs, Bend, OR, #664172 and #664283) cut to generate a linear gradient of shear stress, calculated from (Usami et al., 1993 (link)). Flow was applied for 16 hr in starvation medium. Cells were then fixed with 4% formaldehyde in PBS for 10 min, permeabilized with 0.5% Triton x-100 in PBS for 10 min, blocked with Startingblock buffer (ThermoScientific) for 30 min at room temperature and probed overnight at 4°C with a primary antibody diluted in Startingblock buffer. Slides were stained with Hoechst 33342 to label nuclei, with rabbit anti-p65 antibody (Cell Signaling) to label NF-κB, and with rabbit anti-Smad1 antibody (Cell Signaling).
Baeyens N., Nicoli S., Coon B.G., Ross T.D., Van den Dries K., Han J., Lauridsen H.M., Mejean C.O., Eichmann A., Thomas J.L., Humphrey J.D, & Schwartz M.A. (2015). Vascular remodeling is governed by a VEGFR3-dependent fluid shear stress set point. eLife, 4, e04645.
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2

Western Blot Analysis of Cellular Proteins

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Cultured cells were lysed using RIPA lysis buffer, and total protein (40 μg/sample) was loaded on a 10% sodium dodecyl sulphate polyacrylamide gel. The following primary antibodies were used in this study: rabbit anti‐L‐ferritin (Alpha Diagnostics International), rabbit anti‐phosphor–Smad1/5/8 (Cell Signaling Technology), rabbit anti‐Smad1 antibody (Cell Signaling Technology), rabbit anti‐phosphor–Stat3 (Cell Signaling Technology), rabbit anti‐Stat3 (Cell Signaling Technology) and mouse anti‐β‐actin (Sigma‐Aldrich).
An P., Wang H., Wu Q., Wang J., Xia Z., He X., Wang X., Chen Y., Min J, & Wang F. (2018). Smad7 deficiency decreases iron and haemoglobin through hepcidin up‐regulation by multilayer compensatory mechanisms. Journal of Cellular and Molecular Medicine, 22(6), 3035-3044.
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3

Evaluating Apcdd1 Effects on TGFβ Signaling

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Xenopus apcdd1 cDNA was injected at the one-cell stage at a concentration of 150 ng/μl.
To evaluate the effect of apcdd1 on protein levels of TGFβ receptors, Xenopus embryos were injected in the animal poles of all blastomeres at-4 cell stage with RNAs for alk3-myc (100pg), bmprII-HA (200 pg), alk4-HA (250 pg), and LacZ (1 ng) as loading control, with or without apcdd1-FLAG (2 ng). Animal caps were collected at 4 cell stage in 1 % NP40 buffer, deglycosylated, and run on NuPage 4–12 % gels. Blots were incubated with the same antibodies as above, and with anti-beta-Gal chicken antibodies (1:5,000) and visualized with ECL.
To evaluate the effect of Apcdd1 on endogenous C-terminal phosphorylation of Smad1, 1ng apcdd1 RNA was injected in the animal pole of all blastomeres at the 4-cell stage. Embryos injected with chordin RNA (1 ng) served as controls for the inhibition of BMP-dependent Smad1 phosphorylation. Animal caps were dissected at stage 11 and lysed in the presence of protein phosphatase inhibitor cocktails (Sigma). Western blots were stained with rabbit anti-Smad1 antibody (1:1000, Cell Signaling) and rabbit anti-pSmad1 (C-terminal) antibody (1:2000, from E. Laufer, Columbia U.).
Vonica A., Bhat N., Phan K., Guo J., Iancu L., Weber J.A., Karger A., Cain J.W., Wang E.C., DeStefano G.M., O’Donnell-Luria A.H., Christiano A.M., Riley B., Butler S.J, & Luria V. (2020). Apcdd1 is a dual BMP/Wnt inhibitor in the developing nervous system and skin. Developmental biology, 464(1), 71-87.
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