Cells were lysed in protein lysis buffer (Beyotime). The whole cell lysis products were analyzed with SDS-PAGE and then transferred to PVDF membrane (Millipore). The following primary antibodies were used: anti-KLF4 polyclonal antibody (ab106629, Abcam), anti-DMP1 polyclonal antibody (ab103203, Abcam), anti-DSP polyclonal antibody (NBP191612, NOVUS), and anti-β-ACTIN monoclonal antibody (660091Ig, Proteintech). After incubation with the corresponding antibodies, the membrane was washed 3 times for 5 min each with TBST. We used ImageJ software for further densitometric analysis. Bound primary antibodies were detected by incubating for 1 h with horseradish peroxidase conjugated goat anti-mouse or anti-rabbit IgG (Biofly, China) for analysis. The membrane was washed and developed by a chemiluminescence assay (GE Healthcare).
Nbp191612
Manufactured by Novus Biologicals
NBP191612 is a lab equipment product manufactured by Novus Biologicals. The core function of this product is to perform specific laboratory tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Ab103203, by Abcam (2 mentions)
Chemiluminescence assay, by GE Healthcare (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Ab106629, by Abcam (1 mentions)
β actin, by Proteintech (1 mentions)
Protein lysis buffer, by Beyotime (1 mentions)
Yt5356, by Immunoway (1 mentions)
Sc 393325, by Santa Cruz Biotechnology (1 mentions)
2 protocols using nbp191612
1
Protein Expression Analysis by Western Blot
2
Western Blot Analysis of Osteogenesis Regulators
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Cells were lysed in NP40 Lysis Buffer with protease inhibitor cocktail (MCE). The cell lysis were analyzed with electrophoresis and then transferred to PVDF membrane (Millipore). The filter was blocked for 1-2 hours by 5% nonfat milk in TBST (Tris buffered saline containing Tween 20) and then incubated with the primary antibodies at 4 °C overnight. The primary antibodies used were: anti-IPO7 monoclonal antibody (sc365231, Santa Cruz), anti-RUNX2 polyclonal antibody (YT5356, Immunoway), anti-KLF4 polyclonal antibody (4038, Cell Signaling technology), anti-Phospho-SMAD1/5 monoclonal antibody (9516, Cell Signaling technology), anti-OSX monoclonal antibody (sc393325, Santa Cruz), anti-DLX3 polyclonal antibody (132613AP, Proteintech), anti-DMP1 polyclonal antibody (ab103203, Abcam), anti-DSP polyclonal antibody (NBP191612, NOVUS), anti-β-ACTIN monoclonal antibody (660091Ig, Proteintech) and anti-PCNA polyclonal antibody (102052AP, Proteintech). After incubation with the corresponding antibodies, the filter was washed 3 times for 5 minutes each with TBST. The filter was then incubated with HRP (horseradish peroxidase) conjugated goat anti-Mouse or anti-Rabbit IgG (Biofly, China) for 1 hour. The filter was washed and developed by a chemiluminescence assay (GE Healthcare). We used ImageJ software for further analysis.
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