Following electrophoresis and protein transfer, PVDF membranes were incubated overnight with mouse monoclonal anti-endoglin antibody (1 μg/mL, R&D, USA), mouse monoclonal anti-HA-probe antibody (1 μg/mL, Santa Cruz Biotechnology, USA), mouse monoclonal anti-VE-cadherin antibody (1 μg/mL, Santa Cruz Biotechnology, USA), mouse monoclonal anti-BMPRII antibody (0.5 μg/mL, R&D), mouse monoclonal anti- β-actin antibody (0.2 μg/mL, Sigma), rabbit anti-p-smad1,5,8 (0.5 μg/mL; Santa Cruz Biotechnology, USA) or mouse monoclonal anti-smad 1 antibody (0.5 μg/mL; Santa Cruz Biotechnology, USA). The membranes were washed in Tris-buffered saline with Tween 20, incubated with goat anti-mouse- or goat anti-rabbit polyclonal HRP-conjugated antibodies (0.2 μg/mL; DAKO, USA) for 1 h. Blots were developed in Luminata Crescendo Western HRP substrate (Millipore, USA) and analysed in BioRad ChemiDoc Imager.
Rabbit anti p smad1 5 8
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Rabbit anti-p-smad1,5,8 is a primary antibody that detects phosphorylated forms of SMAD1, SMAD5, and SMAD8 proteins. These proteins are key components of the BMP (Bone Morphogenetic Protein) signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Luminata crescendo western hrp substrate, by Merck Group (1 mentions)
Mouse monoclonal anti β actin antibody, by Merck Group (1 mentions)
Chemidoc imager, by Bio-Rad (1 mentions)
Mouse monoclonal anti ve cadherin antibody, by Santa Cruz Biotechnology (1 mentions)
Lc3ii, by Thermo Fisher Scientific (1 mentions)
Rabbit anti β actin, by Abcam (1 mentions)
Goat anti rabbit igg h l hrp, by Abcam (1 mentions)
Goat anti mouse igg hrp, by Thermo Fisher Scientific (1 mentions)
2 protocols using rabbit anti p smad1 5 8
1
Western Blot Analysis of Endoglin and Related Proteins
2
Investigating OPN, TGF-β, and LC3 in HOb Cells
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HOb cells were seeded and 24 h later samples were added. After three days, cells were collected and lysed. Protein concentrations were measured and sodium dodecyl sulfate polyacrylamide gel electrophoresis was done to separate cell lysates. Lysates were transferred to polyvinylidene difluoride membranes, followed by probing with primary antibodies. Mouse anti-OPN and rabbit-anti TGF-β were purchased from GeneTex Inc. and were used at dilutions 1:1000. Rabbit anti-LC3I and LC3II were purchased from Thermo Fisher Scientific and were used at dilutions 1:1000. Rabbit anti-pSmad 1/5/8 were used at the dilutions of 1:500 and were purchased from Santa Cruz Biotechnology (CA, USA). Rabbit anti-β-actin was purchased from Abcam and was used at dilutions of 1:500. Secondary antibodies, goat anti-mouse IgG HRP from Thermo Fisher Scientific at 1:3000 and goat anti-rabbit IgG H&L HRP from Abcam at 1:3000 were used (Tai et al., 2020 ).
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