After being transfected for 48 h, cells were collected by centrifugation, followed by total protein extraction. The proteins were quantified by BCA (bicinchoninic acid) protein assay kit (Jianglaibio, Shanghai, China) and separated by 12% SDS-PAGE. Then, proteins were transferred to the PVDF membrane and blocked with 5% skim milk. The membranes were incubated with primary antibodies of MMP-2 monoclonal antibody (1 : 500, Invitrogen), MMP-9 polyclonal antibody (1 : 1000, Invitrogen), Cyclin D1 monoclonal antibody (1 : 200, Invitrogen), c-myc monoclonal antibody (1 : 200, Invitrogen), JAK1 monoclonal antibody (1 : 1000, Invitrogen), p-JAK1 polyclonal antibody (1 : 1000, Invitrogen), STAT3 monoclonal antibody (1 : 5000, Invitrogen), p-STAT3 polyclonal antibody (1 : 1000, Invitrogen), and GAPDH monoclonal antibody (1 : 1000, Invitrogen) at room temperature overnight, followed by incubation with the secondary antibody at 37°C for 90 min. Finally, proteins were detected with the ECL method under a multi-imager (Bio-Rap, Hercules, CA, USA).
Gapdh monoclonal antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The GAPDH monoclonal antibody is a laboratory tool used to detect the presence and quantity of the GAPDH protein in biological samples. GAPDH, or Glyceraldehyde 3-phosphate dehydrogenase, is a common housekeeping gene that is essential for cellular metabolism. The antibody can be used in various laboratory techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to study the expression and distribution of GAPDH in different cell types and tissues.
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Lab products found in correlation
C myc monoclonal antibody, by Thermo Fisher Scientific (1 mentions)
Procaspase 3, by Cell Signaling Technology (1 mentions)
Mouse anti cyclin a, by Santa Cruz Biotechnology (1 mentions)
Cleaved caspase 9, by Cell Signaling Technology (1 mentions)
Cyclin b1, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
2 protocols using gapdh monoclonal antibody
1
Protein Expression Analysis by Western Blot
2
Western Blot Quantification of Apoptosis-Related Proteins
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From protein extraction, quantitation, separation by 12 % sodium dodecyl sulfate–polyacrylamide gel electrophoresis, transfer to polyvinylidene fluoride membrane, and to protein visualization, Western blot technology was employed as previously described (Matou-Nasri et al., 2022 (link)). Diluted in blocking buffer, the primary antibodies used were rabbit anti-cleaved caspase-3 (#9664, dilution 1:1000), pro-caspase 3 (#9665, 1:1000), cleaved caspase-9 (#9505, 1:1000), cleaved poly (ADP-ribose) polymerase (PARP) (#5625, 1:500), PARP (#9542, 1:500), mouse pro-caspase-9 (#9508, 1:1000) monoclonal antibodies provided by Cell Signaling Technology (Danvers, MA, USA), and mouse anti-cyclin A (#sc-271645, 1:1000), cyclin B1 (#sc-70898, 1:1000), cyclin D1 (#sc-8396, 1:1000) monoclonal antibodies purchased from Santa Cruz Biotechnology, and mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) monoclonal antibody (#AM4300, 1:5000) from Invitrogen.
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