Cells were lysed with radioimmunoprecipitation assay (RIPA) lysis buffer (C1053, Beijing Applygen Technologies Inc.), and protein concentrations were determined using the bicinchoninic acid (BCA) Protein Assay Kit (PC0020, Solarbio, China). Total protein of 20 μg was separated on a 10% NuPAGETM Bis-Tris Welcome Pack (NP030B, Invitrogen), and the electrophoresed products were transferred to iBlotTM 2 NC Regular Stacks (IB23001, Life Technologies). Membranes were blocked for 30 min at RT using 5% powdered milk, and primary antibodies mouse anti-eGFP mAb (TA-06, ZSGB-BIO, China) and mouse anti-GAPDH mAb (TA-08, ZSGB-BIO, China) were incubated overnight at 4°C. After washing the membranes, the secondary antibody Goat anti-Mouse IgG (H + L) (ZB-2305, ZSGB-BIO, China) was incubated for 2 h at RT. Bands were visualized with Pro-light HRP substrate chemiluminescent system (PA112, Tiangen Biotech Co., Ltd.). Chemiluminescent signals were quantified using ImageJ software.
Goat anti mouse igg h l
Manufactured by ZSGB-BIO
Sourced in China
Goat anti-Mouse IgG (H + L) is a laboratory reagent used for detecting and quantifying mouse immunoglobulins in various immunological applications. It is a polyclonal antibody raised in goats and is reactive to the heavy and light chains of mouse IgG.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti gapdh mab, by ZSGB-BIO (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
Ripa lysis buffer, by Applygen (1 mentions)
Rabbit anti akt, by Abcam (1 mentions)
Rabbit anti pdk1, by Abcam (1 mentions)
Goat anti rabbit igg h l, by ZSGB-BIO (1 mentions)
Rabbit anti mmp 1, by Abcam (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Pvdf membrane, by Abcam (1 mentions)
Bca method, by Thermo Fisher Scientific (1 mentions)
2 protocols using goat anti mouse igg h l
1
Western Blot Analysis of eGFP Expression
2
Protein Quantification and Western Blot Analysis
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Total proteins were extracted, and protein concentrations were quanti ed via the bicinchoninic acid (BCA) method (Thermo Scienti c). A total of 50 µg of protein was separated in 10% SDS-PAGE gels and transferred to PVDF membranes (Millipore, Billerica, MA, USA). After blocking overnight at 4 °C in the 5% BSA, the PVDF membranes were incubated with the primary antibodies, including Rabbit Anti AKT (1/500),Rabbit Anti Caspase-3 (1/5000), Rabbit Anti Caspase-9 (1/2000), Rabbit Anti MDK (1/1000), Rabbit Anti MMP-1 (1/1000), Rabbit Anti PDK1(1/2000)(Abcam, Cambridge, UK), Rabbit Anti p-AKT (Bioss, 1/1000), Rabbit Anti p-PDK1 (1/1000;A nity Biologicals, Ancaster, ON, Canada) and Mouse Monoclonal Anti-GAPDH (1/2000; ZSBIO, Beijing,China) at 4 °C overnight. Horseradish peroxidase-labeled secondary antibodies, including Goat Anti-Mouse IgG(H + L) and Goat Anti-Rabbit IgG(H + L) (ZSBIO), were used at 1:2000 dilution to incubate the membranes at room temperature for 2 h. Signals were detected with the SuperSignal® west pico chemiluminescent substrate (Thermo Fisher Scienti c).
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