Total cell protein was extracted by using radio immunoprecipitation assay (RIPA) lysis buffer. Protein concentration was measured by the BCA Protein Assay Kit (Beyotime Institute of Biotechnology, Nanjing, China). Equal amount of protein samples was used for 10% SDS-PAGE electrophoresis. After that, the protein was transferred onto the PVDF membranes blocked with 5% BSA. These PVDF membranes were added with primary antibodies and incubated overnight at 4°C. After washed with TBST, the secondary antibodies were added to the membranes. The antibodies were as follows: RUNX2 (ab76956, Abcam), BMPR2 (EM1709-92, Huabio), p-SMAD5 (ET1605-5, Huabio), SMAD5 (ET1606-26, Huabio), p-p38 (ER1903-01, Huabio), p38 (ET1602-26, Huabio), and GAPDH (ab181602, Abcam).
Er1903 01
Manufactured by Huabio
Sourced in China
The ER1903-01 is a benchtop centrifuge used for separating particles, cells, or other components from liquid samples. It features a maximum speed of 14,000 RPM and a rotor capacity of up to 4 x 100 mL. The centrifuge is designed for general-purpose laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab76956, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ab181602, by Abcam (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Ab209232, by Abcam (1 mentions)
Et1602 18, by Huabio (1 mentions)
Ab216776, by Abcam (1 mentions)
Ab8226, by Abcam (1 mentions)
Bca protein assay kit, by Elabscience (1 mentions)
Pg212, by Ipsen (1 mentions)
Goat anti mouse igg, by Abcam (1 mentions)
2 protocols using er1903 01
1
Western Blot Analysis of Osteoblast Markers
2
Cardiac Protein Extraction and Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total protein was extracted from frozen heart tissue using a Minute™ muscle tissue total protein extraction kit (SA-06-MS, Invent Biotechnologies, Inc., United States) following the manufacturer’s instructions. Protein samples were quantified using a BCA Protein Assay Kit (E-BC-K318-M, Elabscience, Wuhan, China). Protein samples were resolved on 10% SDS-PAGE gels (PG212, EpiZyme, Shanghai, China) and transferred onto polyvinylidene fluoride membranes (Millipore, Burlington, MA, United States). The membranes were blocked with 5% fat-free milk and incubated overnight at 4°C with primary antibodies (ANP, 1:1000, ab209232, Abcam, United States; β-MHC, 1:1000, 22280-1-AP, Proteintech, China; p38, 1:1000, ET1702-65, Huabio, China; JNK, 1:1000, RT1550, Huabio, China; ERK1/2, 1:1000, 67170-1-Ig, Proteintech, China; p-p38, 1:1000, ER1903-01, Huabio, China; p-JNK, 1:1000, 4668S, Cell Signaling Technology, USA; p-ERK1/2, 1:1000, 4370T, Cell Signaling Technology, United States; DUSP 6, 1:1000, ET1602-18, Huabio, China; β-ACTIN, 1:1000, ab8226, Abcam, United States). Then, the membranes were washed and incubated with secondary antibodies (Goat anti-Mouse IgG, 1:10000, ab216776, Abcam; Goat Anti-Rabbit IgG, 1:10000, ab6721, Abcam). The protein bands were visualized by ECL (Millipore) and quantified using ImageJ software (National Institutes of Health).
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