Total protein quantification assay kit

Manufactured by Nanjing Jiancheng

Sourced in China

The Total Protein Quantification Assay Kit is a laboratory tool used for the quantitative determination of total protein concentrations in various samples. The kit provides a standardized method for measuring the total protein content, which is essential for many applications in biochemistry, cell biology, and related fields.

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Total protein (5 citations) Quantification kits (5 citations)

3 protocols using total protein quantification assay kit

Protein Content and Enzyme Bioactivity Analysis

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Treated mites were washed three times with normal saline (0.9% w/v of NaCl solution). Samples were homogenized in 900 μl of normal saline and centrifuged at 2500 r/min (4 °C) for 10 min. The supernatant was used to determine the water-soluble protein concentration, using a total protein quantification assay kit (A045-3, Nanjing Jiancheng Bioengineering Institute, China). Briefly, the proteins reduced cupric ion to cuprous ion, which was further adducted with bicinchoninic acid (BCA). The product has a maximum absorbance at 562 nm. The protein content was calculated based on the absorbance of the solution.
The enzyme bioactivities of GST, SOD and Ca²⁺-Mg²⁺-ATPase were evaluated using a GST assay kit (catalog no. A004, Nanjing Jiancheng Bioengineering Institute, China), an SOD assay kit (catalog no. A001-3, Nanjing Jiancheng Bioengineering Institute, China) and a Ca²⁺-Mg²⁺-ATPase assay kit (catalog no. A070-3, Nanjing Jiancheng Bioengineering Institute, China) according to the manufacturer’s instructions.

Li L., Lin Z.G., Wang S., Su X.L., Gong H.R., Li H.L., Hu F.L, & Zheng H.Q. (2017). The effects of clove oil on the enzyme activity of Varroa destructor Anderson and Trueman (Arachnida: Acari: Varroidae). Saudi Journal of Biological Sciences, 24(5), 996-1000.

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Antioxidant Enzyme Gene Expression in V. alginolyticus

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Genes encoding antioxidant-related enzymes of SOD, GR and CAT in V. alginolyticus were cloned and sequenced in our previous work, and qPCR primers were designed according to the sequenced results (Table 1), referenced by 16S rDNA gene. Primers were synthesized by Sangon Biological Engineering Technology & Services Co., Ltd. (China). The kits of RNA extraction and reverse transcription were from Beijing TransGen Biotech Co., Ltd. (China). Sodium hydrosulfide (NaSH, H₂S donor), hypotaurine (HT, H₂S scavenger), sodium nitroprusside (SNP, NO donor) and 2-phenyl-4, 4, 5, and 5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO, NO scavenger) came from Sigma-Aldrich Co. LLC (USA), while NOR came from Guangzhou Technology Company Limited (China). ELISA kits for microorganism CBS and NOS, H₂S and NO were provided by Shanghai Jianglai Industrial Ltd (China). Hydrogen Peroxide Assay Kit and Total Protein Quantification Assay Kit were from Nanjing Jiancheng Bioengineering Institute (China).

Chen S., Chang Y, & Ding Y. (2021). Roles of H2S and NO in regulating the antioxidant system of Vibrio alginolyticus under norfloxacin stress. PeerJ, 9, e12255.

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Bicinchoninic Acid Protein Quantification

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Total protein was measured according to the bicinchoninic acid method (with standard sample) (Smith et al., 1985 (link)) with Total Protein Quantification Assay Kit (Nanjing Jiancheng Bioengineering Institute, China). In briefly, the principle of assessment total protein concentration is that Cu⁺ reduced from Cu²⁺ by proteins in the alkaline condition can react with the bicinchoninic acid (BCA) reagent to form the purple complex which can be spectrophotometrically read at 562 nm, and to quantify the protein by comparing with the standard curve. The absorbance is proportional to the protein concentration, so concentration can be obtained following the formula: total protein = (OD_sample- OD_blank)/(OD_standard- OD_blank) × standard sample (524 µg/mL) × sample dilution times.

Chen S., Chang Y, & Ding Y. (2021). Roles of H2S and NO in regulating the antioxidant system of Vibrio alginolyticus under norfloxacin stress. PeerJ, 9, e12255.

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