After protein extraction, protein concentration was determined with a BCA assay (CWBIO, China). A 10% SDS-PAGE gel was loaded with 20 µg of total protein, and the separated proteins were transferred by electroblotting to polyvinylidene fluoride (PVDF) membranes. The membranes were blocked with 5% non-fat dry milk in TBST (50mM tris-buffered saline, pH 7.6, 150mM NaCl, 0.1% Tween 20) and incubated with the primary antibody overnight at 4°C in 5% non-fat dry milk in TBST. Immunolabelling was detected using enhanced chemiluminescence (ECL) reagent (Thermo Fisher Scientific). The antibodies used for Western blot were from the following sources: anti-Runx2 antibody (Abcam, UK; 1:1,000), anti-Osx antibody (Cell Signalling Technology, USA; 1:1,000), anti-ALP antibody (Cell Signalling Technology; 1:1,000), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (Sigma-Aldrich; 1:10,000).
Anti alp antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The Anti-ALP antibody is a laboratory reagent used to detect and study the alkaline phosphatase (ALP) enzyme. ALP is an important enzyme involved in various biological processes. The antibody can be used in techniques such as Western blotting, immunohistochemistry, and ELISA to identify and quantify ALP in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Bca assay, by CWBIO (1 mentions)
Anti glyceraldehyde 3 phosphate dehydrogenase gapdh antibody, by Merck Group (1 mentions)
Anti runx2 antibody, by Abcam (1 mentions)
Ecl reagent, by Thermo Fisher Scientific (1 mentions)
Anti runx2 antibody, by Cell Signaling Technology (1 mentions)
Anti ocn antibody, by Cell Signaling Technology (1 mentions)
Anti skp2 antibody, by Santa Cruz Biotechnology (1 mentions)
Anti foxo1 antibody, by Cell Signaling Technology (1 mentions)
Anti gapdh antibody, by Santa Cruz Biotechnology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
2 protocols using anti alp antibody
1
Western Blot Analysis of Osteogenic Markers
2
Western Blot Analysis of Osteogenic Markers
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Total protein was extracted from tissues or cells using RIPA Lysis Buffer (Beyotime, Beijing, China). The protein was then separated by 12% SDS-PAGE and transferred onto PVDF membranes (Millipore, USA). The membranes were blocked with 5% skim milk and incubated with primary antibodies, including anti-FOXO1 antibody (1:400, Cell Signaling Technology), anti-ALP antibody, anti-Runx2 antibody (1:300, Cell Signaling Technology), anti-Ostx antibody (1:300, Cell Signaling Technology), anti-OCN antibody (1:300, Cell Signaling Technology), anti-Skp2 antibody (1:500, Santa Cruz, USA) and anti-GAPDH antibody (1:800, Santa Cruz). Then, the membranes were incubated with anti-rabbit horseradish peroxidase-conjugate secondary antibody (1:2000, Santa Cruz) at room temperature for 1 h. ECL development solution (Thermo, USA) was used to visualize the proteins.
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