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Rabbit anti smad1 5 8

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The Rabbit anti-Smad1/5/8 is a primary antibody that recognizes the Smad1, Smad5, and Smad8 proteins. These proteins are members of the SMAD family and play a role in the transforming growth factor-beta (TGF-β) signaling pathway.

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Lab products found in correlation

Mouse anti gapdh, by Merck Group (2 mentions) T per protein extraction reagent, by Thermo Fisher Scientific (2 mentions) Rabbit anti phospho smad1 5 8, by Cell Signaling Technology (2 mentions) Mouse anti bmp4, by OriGene (2 mentions) Rabbit ant id2, by Santa Cruz Biotechnology (2 mentions) Rabbit anti noggin, by Merck Group (2 mentions) Halt protease and phosphatase inhibitor, by Thermo Fisher Scientific (2 mentions) Axioplan microscope, by Zeiss (2 mentions) Rabbit anti smad2 3, by Cell Signaling Technology (1 mentions) Fitc conjugated anti rabbit igg, by Merck Group (1 mentions) Rabbit anti mouse actin, by Merck Group (1 mentions) Rabbit anti phospho smad3, by Cell Signaling Technology (1 mentions) Rabbit anti mouse phospho stat1, by Cell Signaling Technology (1 mentions) Facscanto 2, by BD (1 mentions) Facscalibur, by BD (1 mentions)

4 protocols using rabbit anti smad1 5 8

1

Multiparametric Analysis of Signaling Pathways

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Rabbit anti phospho-Smad2 (Millipore, Billerica, MA, USA), rabbit anti Smad2/3 (Cell Signaling Technology, Danvers, MA, USA), rabbit anti phospho-Smad3 (Cell Signaling Technology), goat anti phospho-Smad1/5/8 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti Smad1/5/8 (Santa Cruz Biotechnology), rabbit anti-mouse phospho-Stat1 (Cell Signaling Technology), rabbit anti-mouse ApoA-I (Santa Cruz Biotechnology), rabbit anti-mouse Actin (Sigma, Steinheim, Germany) and HRP conjugate donkey anti-rabbit IgG (Southern Biotech, Birmingham, AL, USA) were used for western blot analysis. Rabbit anti SRB1 monoclonal antibody (Novus Biologicals, CO, USA) and FITC conjugated anti-rabbit IgG (Sigma, Steinheim, Germany) were used for flow cytometry analysis. Fc block and fluorochrome conjugated antibodies against mouse CD45, CD3, CD8, MHC-II, CD11c and TNF-alpha were purchased from BD Biosciences. Intracellular staining for TNF-alpha was performed following manufacturer's instructions. For flow cytometry experiments acquisition was done using either FACSCalibur or FACSCanto II (BD Biosciences). FlowJo Software (TreeStar) was used to analyze cellular events.
Medina-Echeverz J., Fioravanti J., Díaz-Valdés N., Frank K., Aranda F., Gomar C., Ardaiz N., Dotor J., Umansky V., Prieto J, & Berraondo P. (2014). Harnessing High Density Lipoproteins to Block Transforming Growth Factor Beta and to Inhibit the Growth of Liver Tumor Metastases. PLoS ONE, 9(5), e96799.
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2

Protein Expression Analysis of Mouse Dorsal Ganglia

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The DG was micro-dissected from adult mice as previously described.25 The tissue was mechanically homogenized on ice in T-per protein extraction reagent (Thermo Fisher) with Halt protease and phosphatase inhibitors (Thermo Fisher). Lysates were centrifuged at 10,000 rpm and the supernatant was collected for western blot analysis. Western blotting was performed using standard techniques. Primary antibodies used were: mouse anti-BMP4 (1:1000, OriGene, UM500038), mouse anti-GAPDH (1:4000, Millipore, MAB374), rabbit ant-Id2 (1:200, Santa Cruz, sc-489), rabbit anti-noggin (1:2000, Millipore, AB5729), rabbit anti-phospho-Smad1/5/8 (1:1000, Cell Signaling, CS9511), and rabbit anti-Smad1/5/8 (1:1000, Santa Cruz, sc-6031-R). See Supplementary Materials and Methods for detailed procedures.
Brooker S.M., Gobeske K.T., Chen J., Peng C.Y, & Kessler J.A. (2016). Hippocampal bone morphogenetic protein signaling mediates behavioral effects of antidepressant treatment. Molecular psychiatry, 22(6), 910-919.
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3

Protein Expression Analysis of Mouse Dorsal Ganglia

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The DG was micro-dissected from adult mice as previously described.25 The tissue was mechanically homogenized on ice in T-per protein extraction reagent (Thermo Fisher) with Halt protease and phosphatase inhibitors (Thermo Fisher). Lysates were centrifuged at 10,000 rpm and the supernatant was collected for western blot analysis. Western blotting was performed using standard techniques. Primary antibodies used were: mouse anti-BMP4 (1:1000, OriGene, UM500038), mouse anti-GAPDH (1:4000, Millipore, MAB374), rabbit ant-Id2 (1:200, Santa Cruz, sc-489), rabbit anti-noggin (1:2000, Millipore, AB5729), rabbit anti-phospho-Smad1/5/8 (1:1000, Cell Signaling, CS9511), and rabbit anti-Smad1/5/8 (1:1000, Santa Cruz, sc-6031-R). See Supplementary Materials and Methods for detailed procedures.
Brooker S.M., Gobeske K.T., Chen J., Peng C.Y, & Kessler J.A. (2016). Hippocampal bone morphogenetic protein signaling mediates behavioral effects of antidepressant treatment. Molecular psychiatry, 22(6), 910-919.
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4

Immunofluorescent Analysis of SMAD1/5/8

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For immunofluorescence, ATDC5 chondrocytes were plated at 1,000 cells/cm2 on coverslips and grown for 24 hours in normal medium. At 60% confluence, cells were serum deprived for 6 hours and treated with DAPT and/or BMP for 2 hours. Cells were then fixed in 4% paraformaldehyde in PBS for 20 minutes at room temperature and permeabilized with 0.3% Triton X-100 in PBS for 10 minutes. Cells were washed in PBS and incubated with 0.5% BSA dissolved in PBS at room temperature for 20 minutes. Cultures were then incubated for 2 hours at room temperature with rabbit anti-SMAD1/5/8 (Santa Cruz Biotechnology) diluted 1:50 in PBS. After washing with PBS, cells were incubated with FITC conjugated anti-rabbit IgG for 1 hour at room temperature. Reaction controls were performed using a non-immune rabbit immunoglobulin IgG, or by omitting the primary antibody. Cover slips were mounted on slides with PBS/glycerol (1:1), and slides were imaged by fluorescent microscopy using a Zeiss Axioplan microscope.
Shang X., Wang J., Luo Z., Wang Y., Morandi M.M., Marymont J.V., Hilton M.J, & Dong Y. (2016). Notch signaling indirectly promotes chondrocyte hypertrophy via regulation of BMP signaling and cell cycle arrest. Scientific Reports, 6, 25594.
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