Western blotting was used to detect the levels of JKAMP, JNK1, GSK-3β, p-GSK-3β, β-catenin, RUNX2, and OPN. Total protein was isolated from cells using a total protein extraction kit (Keygen Biotech, Nanjing, China). The protein samples were mixed with loading buffer, boiled for 5 min, separated by SDS-PAGE, and transferred to polyvinylidene fluoride membranes [29 (link), 30 (link)]. The membranes were blocked with 5% dry skim milk in Tris-buffered saline with 0.05% (v/v) Tween-20 (TBST) for 1 h and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4 °C overnight. The membrane was washed three times with TBST and then incubated with a secondary labeled anti-rabbit or anti-mouse antibody (1:3000) for 1 h. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
Nbp2 36446ss
Manufactured by Novus Biologicals
Sourced in United States
NBP2-36446SS is a laboratory centrifuge that is capable of separating and concentrating samples. It is designed to operate at high speeds to effectively separate components of varying densities within a sample.
Automatically generated - may contain errors
Lab products found in correlation
Total protein extraction kit, by Keygen Biotech (2 mentions)
Ab92336, by Abcam (2 mentions)
Ab8448, by Abcam (2 mentions)
Ab110724, by Abcam (2 mentions)
Ab181602, by Abcam (2 mentions)
P gsk 3β 5558, by Cell Signaling Technology (2 mentions)
Gsk 3β 12456, by Cell Signaling Technology (2 mentions)
Ab32572, by Abcam (2 mentions)
Enhanced chemiluminescence detection system, by Bio-Rad (2 mentions)
2 protocols using nbp2 36446ss
1
Western Blot Analysis of Key Signaling Proteins
2
Protein Expression Analysis by Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Western blotting was used to detect the levels of JKAMP, JNK1, GSK-3β, p-GSK-3β, β-catenin, RUNX2, and OPN. Total protein was isolated from cells using a total protein extraction kit (Keygen Biotech, Nanjing, China). The protein samples were mixed with loading buffer, boiled for 5 minutes, separated by SDS-PAGE, and transferred to polyvinylidene fluoride membranes [27, 28] . The membranes were blocked with 5% dry skim milk in Tris-buffered saline with 0.05% (v/v) Tween-20
(TBST) for 1 hour and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4°C overnight. The membrane was washed three times with TBST and then incubated with a secondary labelled anti-rabbit or anti-mouse antibody (1:3000) for 1 hour. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
(TBST) for 1 hour and then incubated with primary antibodies against JKAMP (NBP2-36446SS) (Novus, Littleton, USA), GAPDH (ab181602), JNK1 (ab110724), β-Catenin (ab32572), RUNX2 (ab92336) and OPN (ab8448) (Abcam, Cambridge, UK), GSK-3β (12456), or p-GSK-3β (5558) (Cell Signaling Technology, Danvers, USA) at 4°C overnight. The membrane was washed three times with TBST and then incubated with a secondary labelled anti-rabbit or anti-mouse antibody (1:3000) for 1 hour. The membrane was then washed three times with TBST and developed using an enhanced chemiluminescence detection system (Bio-Rad, Hercules, USA).
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