Proteins were extracted by radioimmunoprecipitation assay (RIPA) Lysis buffer (Abcam, Cambridge, MA, USA) and quantified by bicinchoninic acid (BCA) Protein Assay Kit (Thermo Scientific, Rockford, IL, USA). 40 µg of protein per sample was separated onto 10% SDS-PAGE gels and transferred onto polyvinyl difluoride (PVDF) filter membrane (Bio-Rad, Hercules, CA, USA). Then, PVDF membranes were blocked with 5% non-fat dried milk for 2 h, and incubated with anti-BZW1 (1: 1000, ABcam Corp, USA), anti-Smad1 (1: 1000, ABcam Corp, USA), anti-p-Smad1 (1: 1000, ABcam Corp, USA), anti-Smad3 (1: 1000, ABcam Corp, USA), anti-p-Smad3 (1: 1000, ABcam Corp, USA), anti-TGF-β1 (1: 2000, ABcam Corp, USA) and anti-GAPDH (1: 2000, ABcam Corp, USA), respectively at 4 overnight. Finally, membranes were incubated with secondary antibodies (horseradish peroxidase-conjugated anti-rabbit). Signals were developed using an enhanced chemiluminescence reaction kit (Applygen Technologies, Beijing, China).
Anti smad1
Manufactured by Abcam
Sourced in United States
Anti-Smad1 is a laboratory reagent that can be used to detect the Smad1 protein. Smad1 is a key mediator in the transforming growth factor-beta (TGF-β) signaling pathway and plays a role in various cellular processes. The Anti-Smad1 product can be used in techniques such as Western blotting to study the expression and regulation of the Smad1 protein.
Automatically generated - may contain errors
Lab products found in correlation
Anti gapdh, by Abcam (2 mentions)
Anti p smad1, by Abcam (2 mentions)
Anti β actin, by Merck Group (2 mentions)
Anti ocn, by Abcam (2 mentions)
Anti runx2, by Abcam (2 mentions)
Anti opn, by Abcam (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Enhanced chemiluminescence reaction kit, by Applygen (1 mentions)
Pvdf filter membrane, by Bio-Rad (1 mentions)
Ripa lysis buffer, by Abcam (1 mentions)
Anti smad3, by Abcam (1 mentions)
Anti p smad3, by Abcam (1 mentions)
Anti tgf β1, by Abcam (1 mentions)
Anti akt, by Cell Signaling Technology (1 mentions)
Anti erk, by Cell Signaling Technology (1 mentions)
Tris hcl, by Merck Group (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti p smad1 5, by Cell Signaling Technology (1 mentions)
Fitc conjugated anti mouse antibody, by ABclonal (1 mentions)
Anti bmp2, by Wuhan Servicebio Technology (1 mentions)
6 protocols using anti smad1
1
Western Blot Analysis of BZW1, Smad, and TGF-β1
2
Antibody-based Analysis of Cellular Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies were used in this study: monoclonal anti‐Smad1/5 (Cell Signaling Technology, 6944); anti‐pSmad1/5 (Cell Signaling Technology, 9516); anti‐Smad1 (Abcam, ab63356); anti‐Keratin 18 (CK18) (MutiSciences, 70‐ab36769‐050); anti‐p‐Akt (Cell Signaling Technology, 4060); anti‐Akt (Cell Signaling Technology, 2920); anti‐p‐Erk (Cell Signaling Technology, 4370); anti‐Erk (Cell Signaling Technology, 4695); anti‐Osteocalcin (OCN) (Biorbyt, orb259644); anti‐Ki67‐FITC (fluorescein isothiocyanate) (Biolegend, 652 409); anti‐BMP2 (Servicebio, GB11252); TRITC (Tetramethylrhodamine)‐conjugated anti‐Rabbit antibody (abclonal, AS040); FITC‐conjugated anti‐Mouse antibody (abclonal, AS001) and anti‐β‐Actin (Sigma, A1978). Reagents: BMP2 (R&D, 355‐BM‐100); 4',6‐diamidino‐2‐phenylindole (DAPI) (Solarbio, C0060) and carboxyfluorescein succinimidyl amino ester (CFSE) (eBioscience, 65‐0850‐84). Tris‐HCl, NaCl and other chemicals were from Sigma.
3
Osteoblast Differentiation Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proteins lysates from MC3T3-E1 cells were partitioned by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a PVDF membrane (EMD Millipore, Billerica, MA, USA). Blotting was carried out with spectific antibodies [anti-B-cell lymphoma-2 (Bcl-2) associated X protein (Bax), 1:5,000 dilution, cat. no. ab32503; Abcam; anti-Bcl-2, 1:1,000 dilution, cat. no. ab692; Abcam; anti-OPN, 1:1,000 dilution, cat. no. ab8448; Abcam; anti-OCN, 1:1,000 dilution, cat. no. ab13418; Abcam; anti-Osterix, 1:1,000 dilution, cat. no. ab94744; Abcam; anti-BMP2, 1:500 dilution, cat. no. ab14933; Abcam; anti-Smad1, 1:1,000 dilution, cat. no. ab33902; Abcam; anti-Smad5, 1:1,000 dilution, cat. no. ab194661; Abcam; anti-Smad8, 1:5,000 dilution, cat. no. ab13723; Abcam; anti-RunX2, 1:500 dilution, cat. no. ab23981; Abcam; anti-β-actin, 1:1,000 dilution, cat. no. ab8227; Abcam]. After that, horseradish peroxidase-conjugated secondary antibodies (bs-0293M; Bioss, Beijing, China) were added and maintained at room temperature for 1 h. The results were assessed by enhanced chemiluminescent reagents (EMD Millipore) using an ECL system (Amersham Pharmacia, Piscataway, NJ, USA).
4
Bone Marrow Stromal Cell Protein Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMSCs were lysed using RIPA buffer (Sigma-Aldrich). Following detecting protein concentration, cell lysate was isolated using 10% SDS-PAGE, followed by being transferred on PVDF membranes. After incubating with primary antibodies (anti-BGLAP, anti-COL1A1, anti-RUNX2, anti-OCN, anti-OPN, anti-METTL14, anti-FTO, anti-ALKBH5, anti-WTAP, anti-RBM15, anti-VIRMA, anti-METTL3, anti-SMAD1, and anti-GAPDH, Abcam, Cambridge) at 4 °C overnight, the membranes were incubated with HRP-conjugated secondary antibody (Abcam) at 25 °C for 1 h. Bands were visualized using immobilon ECL ultra western HRP substrate (Sigma-Aldrich). The original bands are shown in the Supplementary material .
5
Comprehensive Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot assays were performed as described previously,39 (link) using anti-ANP, anti-β-MHC, anti-α-SMA, anti-p-Smad1, anti-Smad1, anti-SRF, anti-E2F2, anti-PGC-1α, anti-NRF2 (Abcam), anti-ACTA1, anti-COL1A1, anti-COL3A1, anti-TRX2, anti-SOD2, anti-HO-1, anti-SRF (Proteintech), anti-SIRT1, anti-p-Smad3, anti-Smad3, anti-p-NF-кB p65, anti-NF-кB p65 (Cell Signaling Technology), anti-RB1or anti-GAPDH (Santa Cruz Biotechnology).
6
Detailed Western Blot Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed as described previously 63 (link), using the following antibodies: anti-NGFR, Cell Signaling, catalog # 8238S; anti-phospho-NF-κB p65 (Ser536), Cell Signaling, catalog #3033; anti-NF-κB p65 (D14E12), Cell Signaling, catalog #8242; anti-phospho-IκBα (Ser32), Cell Signaling, catalog # 2859; anti-IκBα, Novus Biologicals, catalog # NB100–56507; anti-p-SMAD1, Cell Signaling, catalog # 9516S; anti-SMAD1, abcam, catalog # ab63356; anti- Phospho-IKKα (Ser176)/IKKβ (Ser177), Cell Signaling, catalog #2078; anti-IKKβ, Cell Signaling, catalog #2678; anti-β-actin, Sigma-Aldrich, catalog # A5441; anti-RANKL, Novus Biologicals, Clone 12A668.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!