Total proteins were extracted using radio-immunoprecipitation assay cell lysis buffer (Beyotime). Afterward, the proteins were performed with 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis for 2 h, transferred onto membranes, and blocked with 5% skim milk powder for 2 h, and then incubated with primary antibodies EIF4G2, vascular endothelial growth factor (VEGF), and glyceraldehyde phosphate dehydrogenase (GAPDH) (all 1:1000 and from Santa Cruz Biotechnology, Inc, CA, USA). Relative secondary antibody was added for reaction and enhanced chemiluminescent method was applied for development. The gray values of protein band was assessed with GAPDH as the loading control, and then the relative protein expression was calculated.
Eif4g2
Manufactured by Santa Cruz Biotechnology
Sourced in United States
EIF4G2 is a protein involved in the initiation of protein synthesis. It functions as a scaffold that coordinates the assembly of the translation initiation complex. EIF4G2 plays a critical role in the regulation of mRNA translation.
Automatically generated - may contain errors
Lab products found in correlation
Radio immunoprecipitation assay cell lysis buffer, by Beyotime (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Sds page gel, by Beyotime (1 mentions)
Ripa reagent, by Thermo Fisher Scientific (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Ecl detection system, by Thermo Fisher Scientific (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
2 protocols using eif4g2
1
Protein Expression Analysis by Western Blot
2
Protein Expression Analysis in Chondrocytes
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The transfected chondrocytes were washed with cold PBS and lysed in RIPA reagent (Invitrogen, Carlsbad, CA, U.S.A.). The proteins were quantified using the BCA Protein Assay kit (Pierce Biotechnology, Rockford, IL, U.S.A.) and electrophoresed on a 10% SDS-PAGE gel (Beyotime Biotechnology). Then, the proteins were transferred to polyvinylidene fluoride membranes (Millipore, Bedford, MA, U.S.A.), and blocked with 5% nonfat milk for 2 h at 37°C. The membranes were incubated with primary antibodies against EIF4G2 (Santa Cruz Biotechnology, Santa Cruz, CA, U.S.A.) and GPAHD (Santa Cruz Biotechnology) at 4°C overnight. All primary antibodies were diluted in 1% bovine serum albumin (Beyotime Biotechnology) solution with a dilution of 1:2000. After the membranes were washed with TBST buffer, and incubated with HRP-conjugated secondary antibody (Santa Cruz Biotechnology). Signals of bands were recorded using ECL detection system (Pierce Biotechnology) according to manufacturer’s information. Data were analyzed using Quantity One software (Bio-Rad, Hercules, CA, U.S.A.).
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