Bradford assay kit

Manufactured by Sangon

Sourced in China

The Bradford assay kit is a colorimetric protein quantification method used to determine the concentration of proteins in a sample. It utilizes the binding of the dye Coomassie Brilliant Blue G-250 to proteins, resulting in a color change that can be measured spectrophotometrically.

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Lab products found in correlation

Ni nta sepharose column, by GE Healthcare (1 mentions) 0.22 μm filter, by Merck Group (1 mentions) Amicon ultra centrifugal filter, by Merck Group (1 mentions) Protease inhibitor cocktail, by Transgene (1 mentions) Ni nta column, by Yeasen (1 mentions) T4 ligase, by Takara Bio (1 mentions) Pgex 4t 2, by Thermo Fisher Scientific (1 mentions) Restriction enzyme, by Takara Bio (1 mentions) Ptk787, by MedChemExpress (1 mentions) Taq dna polymerase, by Takara Bio (1 mentions) E coli dh5α, by Thermo Fisher Scientific (1 mentions) Isopropyl β d 1 thiogalactopyranoside iptg, by Merck Group (1 mentions) Ampicillin, by Merck Group (1 mentions)

Close spelling variants of this product

C503041-1000 Modified Bradford Protein Assay Kit Modified Bradford Protein Assay Kit Bradford protein assay kit Bradford bioassay-Bradford protein assay kit Bradford assay Bradford kit

6 protocols using bradford assay kit

Purification and Characterization of rSjTGR-Sec and wtSjTGR

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The rSjTGR-Sec was expressed and harvested as described previously [13 (link), 19 (link)]. Most of rSjTGR-Sec was expressed in E. coli as a soluble His-tagged fusion protein when bacterial growth occurred at 24 °C. The rSjTGR-Sec was purified by His Mag™ Agarose beads according to manufacturer’s instructions. The concentration of purified rSjTGR-Sec was determined using a Bradford assay kit (Sangon, China), and the protein was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and stored at 4 °C [20 (link)].
The SWAP containing wtSjTGR was prepared from adult schistosomes. Rabbits were infected with cercariae of S. japonicum by exposing their abdominal skin and were sacrificed at 42 days after infection to collect the adult schistosomes by perfusion. The worms were washed three times with phosphate-buffered saline (PBS). After sufficient grinding of the adult schistosomes in PBS, a suitable amount of benzoyl sulfonyl fluoride (PMSF) solution was added to obtain a final concentration of 1 mM. The homogenate was centrifuged at 4 °C, 12,000×g for 20 min [1 (link), 10 (link)]. The concentration of supernatant containing wtSjTGR was determined by Bradford assay kit (Sangon, China).

Li G., Guo Q., Feng C., Chen H., Zhao W., Li S., Hong Y, & Sun D. (2021). Synthesis of oxadiazole-2-oxide derivatives as potential drug candidates for schistosomiasis targeting SjTGR. Parasites & Vectors, 14, 225.

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Purification of Recombinant Hybrid Peptide

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Recombinant hybrid peptide was purified by Ni-NTA Sepharose column (GE Healthcare, Chicago, IL, USA). The expressed culture medium centrifuged at 10,000× g for 15 min, 4 °C, and the supernatant was collected and then filtrated by 0.22 μm filter (Millipore, Burlington, MA, USA). Ni-NTA Sepharose column was pre-equilibrated with buffer containing 20 mM Tris-HCl, 300 mM NaCl, 40 mM imidazole, pH 7.5. The supernatants were loaded onto Ni-NTA column and washed with equilibration buffer. The bound peptide then eluted by buffer (20 mM Tris-HCl, 300 mM NaCl, 40 mM imidazole, pH 7.5) five times. Fractions were pooled and then concentrated by Amicon Ultra centrifugal filters (Millipore, Burlington, MA, USA). The collected peptide was further purified by reversed-phase high performance liquid chromatography (RP-HPLC). Purified peptide was quantified through Bradford assay kit (Sangon Biotech, Shanghai, China) using bovine serum albumin as a standard and analyzed by 16%/6 M urea Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Tricine-SDS-PAGE) [37 (link)].

Li Z., Cheng Q., Guo H., Zhang R, & Si D. (2020). Expression of Hybrid Peptide EF-1 in Pichia pastoris, Its Purification, and Antimicrobial Characterization. Molecules, 25(23), 5538.

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Peach Protein Extraction Protocol

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Protein extraction was carried out as described previously.¹⁸ Briefly, 1 g of homogenized material was ground to powder in liquid nitrogen and taken up in Coca’s solution (0.1 M Na₂CO₃, 0.1 M NaHCO₃, 0.1 M NaCl, 2 mM EDTA–Na₂, 20 mM sodium diethyldithiocarbamate trihydrate), at 1:5 (wt/vol) for pulp and whole samples and 1:10 (wt/vol) for the peel samples. Each peach variety contained three replicate samples. The supernatant was stored at −30°C and used for Pru p 1 quantification within 2 days. Total protein content was determined with the Bradford Assay Kit (Sangon Biotech) according to the manufacturer’s instructions.

Jin J., Gan K., Zhao L., Jia H., Zhu Y., Li X., Yang Z., Ye Z., Cao K., Wang Z., Yu M., Zhang Y., Ma Z., Liu H., Arús P., Akkerdaas J.H., Gao Z, & van Ree R. (2021). Peach allergen Pru p 1 content is generally low in fruit but with large variation in different varieties. Clinical and Translational Allergy, 11(3), e12034.

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Exosome Isolation and Quantification

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Exosomes were isolated as previously described 67 (link). Briefly, cell culture medium was sequentially centrifuged at 1000×g for 10 min, followed by 10,000×g for 30 min. The supernatant was collected and filtered with a 0.22 µm filter (Millex, Germany), followed by ultracentrifugation at 100,000×g for 1 h. Exosome pellets were washed in a large volume of PBS and recovered by centrifugation at 100,000×g for 1 h and was resuspended in sterile PBS for the following experiments. The protein concentration of isolated exosomes was quantified by using the Bradford assay kit (Sangon Biotech, China).

Dong X., Lei Y., Yu Z., Wang T., Liu Y., Han G., Zhang X., Li Y., Song Y., Xu H., Du M., Yin H., Wang X, & Yan H. (2021). Exosome-mediated delivery of an anti-angiogenic peptide inhibits pathological retinal angiogenesis. Theranostics, 11(11), 5107-5126.

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Large-Scale Recombinant Protein Expression

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For large scale expression, 10 mL overnight culture was inoculated into 400 mL LB media containing 100 µg/mL Kanamycin and incubated at 37 °C at 220 rpm. When OD₆₀₀ reached 0.6–0.8, protein expression was induced with the final concentration of 0.5 mM of isopropyl β-d-1-thiogalactopyranoside (IPTG). After incubation at 16 °C for an additional 20 h, bacterial pellet was collected by centrifugation and resuspended in Equilibration buffer (200 mM NaCl, 50 mM Tris-HCl, 10 mM imidazole, pH 8.0) with protease inhibitor cocktail (Transgen, Beijing, China). Bacterial disruption was performed in a high-pressure homogenizer for optimal lysis on ice. Supernatant was collected after centrifugation and filtered through a 0.22 μm filter. After the selective binding of the protein onto a Ni-NTA column (Yeasen, Shanghai, China), elution was achieved by applying a linear gradient of Elution buffer (200 mM NaCl, 50 mM Tris-HCl, 10 mM-500 mM imidazole, pH 8.0). The eluted samples were then evaluated by SDS-PAGE and quantified using the Bradford Assay Kit (Sangon Biotech, Shanghai, China).

Liu Z.H., Deng Z.F., Lu Y., Fang W.H, & He F. (2022). A modular and self-adjuvanted multivalent vaccine platform based on porcine circovirus virus-like nanoparticles. Journal of Nanobiotechnology, 20, 493.

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Pancreatic Cancer Protein Expression Analysis

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The tissue of pancreatic cancer was provided by the Shanghai First Peoples Hospital. Trizol for extracted RNA was obtained from Invitrogen (Shanghai, China). Restriction enzymes, T4 ligase, and Taq DNA Polymerase were purchased from TaKaRa (Dalian, China). The kits for the PCR product cleaning, DNA recovery from the gel after electrophoresis, and plasmid extraction were purchased from Sangon (Shanghai, China). E. coli DH5α from Invitrogen and E. coli BL21 (DE3) from Novagen (Shanghai, China) were used for the plasmid propagation and protein overexpression, respectively. The plasmid pGEX-4T-2 was purchased from Invitrogen. The Bradford assay kit for measuring protein concentrations was purchased from Sangon. The anti-VEGFR-2 antibody and mouse monoclonal antibody against phosphotyrosine (PY99) were purchased from BD (Becton, Dickinson and Company, Shanghai, China). All purified chromatography columns were obtained from GE Healthcare (Shanghai, China). VEGFR-2 expressed in Sf9 (Spodoptera frugiperda) insect cells was purchased from Abcam (Shanghai, China). PTK787, a known VEGFR-2 inhibitor, was obtained from MedChemExpress (Shanghai, China). The isopropyl β-D-1-thiogalactopyranoside (IPTG) and ampicillin were obtained from Sigma-Aldrich (Shanghai, China). All the solutions were made up with MilliQ water.

Wei J., Cao X., Zhou S., Chen C., Yu H., Zhou Y, & Wang P. (2015). Soluble Expression and Purification of the Catalytic Domain of Human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli. Journal of microbiology and biotechnology, 25(8).

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