Bradford protein assay

Manufactured by Takara Bio

Sourced in China, Japan

The Bradford protein assay is a colorimetric biochemical assay used to quantify the total protein concentration in a sample. It is a fast and reliable method for determining protein levels by measuring the absorbance of the sample at a specific wavelength.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Bradford Protein Assay Kit (41 citations) TaKaRa Bradford Protein Assay Kit (8 citations) Protein assay kit for the Bradford method (2 citations)

6 protocols using bradford protein assay

Comprehensive Proteomic Sample Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Samples were minced and lysed in lysis buffer (8 M urea, 100 mM Tris hydrochloride, pH 8.0) containing protease and phosphatase inhibitors (Thermo Scientific) and then sonicated for 1 min (3 s on and 3 s off, amplitude 25%, SONICS, VCX130). The lysates were centrifuged at 14,000×g for 10 min and supernatants were collected as whole-tissue extracts. Protein concentrations were determined using the Bradford protein assay (TaKaRa, T9310A). Extracts (50 μg protein) were reduced with 10 mM dithiothreitol at 56 °C for 30 min and alkylated with 10 mM iodoacetamide at room temperature in the dark for 30 min. The samples were digested with trypsin using a filter-aided sample preparation method^{57 (link)}. Tryptic peptides were separated using a home-made reverse-phase C18 column. Peptides were eluted, vacuum-dried (Concentrator Plus, Eppendorf), and analyzed by liquid chromatography-tandem MS (LC-MS/MS).

Zhang H., Bai L., Wu X.Q., Tian X., Feng J., Wu X., Shi G.H., Pei X., Lyu J., Yang G., Liu Y., Xu W., Anwaier A., Zhu Y., Cao D.L., Xu F., Wang Y., Gan H.L., Sun M.H., Zhao J.Y., Qu Y., Ye D, & Ding C. (2023). Proteogenomics of clear cell renal cell carcinoma response to tyrosine kinase inhibitor. Nature Communications, 14, 4274.

+ Open protocol

+ Expand

Western Blot Analysis of Renal Cell Carcinoma

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins were extracted from RCC cells using cell lysis buffer, and protein concentration was determined using a Bradford protein assay (Takara Biotechnology, Dalian, P.R. China). Equal amounts of protein samples were loaded and isolated using 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and blotted onto PVDF membranes (Millipore Corp., Billerica, MA, USA). After being blocked by 5% nonfat milk, the membranes were incubated with primary antibodies at 4°C overnight. The following primary antibodies were used: anti-TPD52, anti-E-cadherin, anti-N-cadherin, anti-vimentin, anti-phospho-PI3K, anti-PI3K, anti-phospho-Akt, anti-Akt, and anti-GAPDH (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Subsequently, the film was incubated with horseradish peroxidase-labeled secondary antibody at room temperature for 1 h. The immune-reactive protein bands were visualized by the ECL kit (Pierce, Rockford, IL, USA). Densitometric analysis was performed using Image-Pro Plus software (Media Cybernetics, Silver Spring, MD, USA).

Zhao Z., Liu H., Hou J., Li T., Du X., Zhao X., Xu W., Xu W, & Chang J. (2017). Tumor Protein D52 (TPD52) Inhibits Growth and Metastasis in Renal Cell Carcinoma Cells Through the PI3K/Akt Signaling Pathway. Oncology Research, 25(5), 773-779.

+ Open protocol

+ Expand

Protein Expression Analysis of EC Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

EC cells were harvested and washed twice with PBS and then lysed in RIPA buffer (Cell Signaling Technology, Danvers, MA, USA) with protease and phosphatase inhibitors on ice for 10 min. The protein concentration was determined using a Bradford protein assay (Takara Biotechnology, Dalian, P.R. China). Equal amounts of protein sample were separated by 10% SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Boston, MA, USA). Proteins on PVDF membranes were blocked with a 5% skim milk solution and then incubated with various primary antibodies [DDX5, E-cadherin, N-cadherin, vimentin, β-catenin, c-Myc, and cyclin D1 or GAPDH from Santa Cruz Biotechnology (Santa Cruz, CA, USA)] at 4°C overnight. Subsequently, membranes were washed extensively with TBST and incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody (Santa Cruz Biotechnology) for 1 h at room temperature. The membranes were visualized using enhanced chemiluminescence (ECL) reagent. Developed films were digitized by scanning, and the optical densities were analyzed using the ImageJ software.

Ma Z., Feng J., Guo Y., Kong R., Ma Y., Sun L., Yang X., Zhou B., Li S., Zhang W., Jiang J., Zhang J., Qiao Z., Cheng Y., Zha D, & Liu S. (2017). Knockdown of DDX5 Inhibits the Proliferation and Tumorigenesis in Esophageal Cancer. Oncology Research, 25(6), 887-895.

+ Open protocol

+ Expand

Protein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total protein extracts were lysed in lysis buffer [20 mM HEPES (pH 7.6), 350 mM NaCl, 20% glycerol, 0.5 mM EDTA, 0.1 mM EGTA, 1% NP-40, 50 mM NaF, 0.1 mM DTT, 0.1 mM PMSF, and a protease inhibitor cocktail]. The protein concentration was determined using a Bradford protein assay (Takara Biotechnology, Dalian, China). Equal amounts of protein were electrophoresed on SDS-PAGE and blotted onto PVDF membranes (Millipore Corp., Billerica, MA, USA). The membranes were blocked in 5% nonfat dry milk for 2 h and incubated with various primary antibodies (anti-SASH1, anti-E-cadherin, anti-N-cadherin, anti-phospho-PI3K, anti-PI3K, anti-phospho-Akt, anti-Akt or anti-β-actin; from Santa Cruz Biotechnology, Santa Cruz, CA, USA) overnight at 4°C, followed by horseradish peroxidase (HRP)-conjugated IgG (Santa Cruz Biotechnology). The immunoreactive protein bands were visualized by ECL kit (Pierce, Rockford, IL, USA).

Zong W., Yu C., Wang P, & Dong L. (2016). Overexpression of SASH1 Inhibits TGF-β1-Induced EMT in Gastric Cancer Cells. Oncology Research, 24(1), 17-23.

+ Open protocol

+ Expand

Western Blot Analysis of MAD2L1 Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in RIPA buffer (Nacalai Tesque, Kyoto, Japan) and centrifuged at 15,000rpm for 10 min at 4 °C. The resulting supernatants were collected and the protein concentration was determined using the Bradford Protein Assay (Takara Bio Inc., Shiga, Japan). The proteins were separated on 12% Mini-PROTEAN TGX Precast Gels (Bio-Rad, CA, USA) and transferred to 0.2-µm polyvinylidene difluoride membranes (Bio-Rad). After blocking for 1 h at room temperature with ECL Prime Blocking Agent (Cytiva, Tokyo, Japan), the membranes were incubated with a primary antibody against MAD2L1 (1:500) (#10337-1-AP; Proteintech Group, Inc, IL, USA) for 16 h at 4 °C. A primary antibody against β-actin (1:2000) (Santa Cruz Biotechnology) was used for normalization. After washing, mouse and rabbit peroxidase-linked secondary antibodies (both from Cytiva) were incubated with the membranes for 1 h at room temperature. The protein signals were detected using an ECL Prime Western blotting detection reagent (Cytiva) and quantified using ImageQuant 500 (Cytiva). The intensity of the protein bands was compared using ImageJ software and the experiments were performed in triplicate.

Makinoya M., Miyatani K., Matsumi Y., Sakano Y., Shimizu S., Shishido Y., Hanaki T., Kihara K., Matsunaga T., Yamamoto M., Tokuyasu N., Takano S., Sakamoto T., Hasegawa T., Saito H., Nakayama Y., Osaki M., Okada F, & Fujiwara Y. (2024). Exosomal miR-493 suppresses MAD2L1 and induces chemoresistance to intraperitoneal paclitaxel therapy in gastric cancer patients with peritoneal metastasis. Scientific Reports, 14, 10075.

+ Open protocol

+ Expand

Quantifying RsmA and RsmE Protein Levels

Check if the same lab product or an alternative is used in the 5 most similar protocols

To measure the protein levels of RsmA-FLAG and RsmE-FLAG, P. fluorescens cells containing the FLAG tag were cultured in LB at 28 °C for 12 h and 1-ml samples were taken. Cells were then suspended in phosphate-buffered saline (PBS) buffer and lysed by sonication. The protein in crude lysates was quantified using the Bradford protein assay (TaKaRa). Total proteins were subjected to SDS-PAGE gel electrophoresis and transferred onto PVDF membrane (Millipore). Blots were washed with PBS containing 0.05% Tween-20 and probed with rabbit-anti-FLAG antibody (Cowin-Biotech, Beijing, China) as primary antibody and mouse-anti-RNAP antibody as the loading control. The resulting bots were incubated for 1 min in chemiluminescence (ECL) reagent using the eECL Western Blot kit (Cowin-Biotech, Beijing, China) and the proteins bands were detected on the X-ray film.

Zhang B., Zhao H., Wu X, & Zhang L.Q. (2020). The Oxidoreductase DsbA1 negatively influences 2,4-diacetylphloroglucinol biosynthesis by interfering the function of Gcd in Pseudomonas fluorescens 2P24. BMC Microbiology, 20, 39.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!