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Bicinchoninic acid

Bicinchoninic acid (BCA) is a chemical compound used in colorimetric assays to quantify the total protein concentration in a sample.
It works by chelating copper ions, which are then reduced by the proteins present, resulting in a purple-colored complex that can be measured spectrophotometrically.
The BCA assay is a sensitive, reproducible, and widely-used method for protein quantification in biochemistry and cell biology research.
Optimizing the BCA assay protocl can boost research productivity and reproducibility, which is where PubCompare.ai's AI-driven platform comes in handy by identifying the most effective and reoducible BCA assay methods from literature, preprints, and patents.

Most cited protocols related to «Bicinchoninic acid»

1
Quantitative Global Lipid Analysis
Cited 24 times
Mice (male, C57 BL/J background, 4 months of age) were purchased from The Jackson Laboratory (Bar Harbor, ME, USA). All animal procedures were performed in accordance with the Guide for the Care and Use of Laboratory Animals and were approved by the Animals Studies Committee at Washington University. Mice were anesthetized by asphyxiation with CO2 and tissue samples were immediately harvested and freeze-clamped at the temperature of liquid nitrogen. The tissue wafers were pulverized into a fine powder with a stainless-steel mortar and pestle. A sample (approximately 10 mg) from each powdered tissue sample was weighed and homogenized in 1 mL of ice-cooled diluted (0.1×) phosphate-buffered saline with a Potter-Elvehjem tissue grinder. Protein assays on each individual homogenate were performed using a bicinchoninic acid protein assay kit (Pierce, Rockford, IL, USA) with bovine serum albumin as a standard. After a certain amount of homogenate (~1 mL) from each sample had been transferred to a disposable culture borosilicate glass tube (16 × 100 mm), 4 mL of CHCl3/MeOH (1:1, v/v) and the proper volume of LiCl solution to make up to 1.8 mL of final LiCl solution (50 mM) were added to each individual test tube. Internal standards in a pre-mixed solution containing one species of a class of interest for quantitative global lipid analysis of individual molecular species of lipid classes were also added to each tissue sample based on protein concentration prior to extraction of lipids. Thus, the lipid content could be normalized to the protein content and quantified directly through ion intensity comparison relative to that of a selected internal standard (i.e., ratiometric comparison). The selection of the proper internal standards for quantitation has been extensively discussed previously.4 (link) The selected internal standards only represent ≪ 1% of endogenous cellular lipid molecular species present as demonstrated by ESI-MS lipid analysis without addition of these internal standards. Each individual lipid extract was reconstituted with a volume of 500 μL/mg of tissue protein in 1:1 CHCl3/MeOH. The lipid extracts were finally flushed with nitrogen, capped, and stored at −20°C for ESI-MS (typically analyzed within 1 week) as previously described.21 (link)
Han X., Yang K, & Gross R.W. (2008). Microfluidics-based electrospray ionization enhances the intrasource separation of lipid classes and extends identification of individual molecular species through multi-dimensional mass spectrometry: development of an automated high-throughput platform for shotgun lipidomics. Rapid communications in mass spectrometry : RCM, 22(13), 2115-2124.
Publication 2008
Animals Animals, Laboratory Asphyxia bicinchoninic acid Biological Assay Cells Chloroform Freezing Lipids Males Mice, House Nitrogen Phosphates Proteins Saline Solution Serum Albumin, Bovine Stainless Steel Staphylococcal Protein A Tissues
2
Robust Quantification of Breast Cancer Proteins
Cited 21 times
RPPA was completed independently and at different time points for training and tests sets using individual arrays. Protein was extracted from human tumors and RPPA was performed as described previously [16 -19 (link)]. Lysis buffer was used to lyse frozen tumors by homogenization (excised tumors) or sonication (FNAs). Tumor lysates were normalized to 1 μg/μL concentration as assessed by bicinchoninic acid assay (BCA) and boiled with 1% SDS. Supernatants were manually diluted in five-fold serial dilutions with lysis buffer. An Aushon Biosystems 2470 arrayer (Burlington, MA) created 1,056 sample arrays on nitrocellulose-coated FAST slides (Schleicher & Schuell BioScience, Inc.). Slides were probed with 146 validated primary antibodies (Additional File 1, Table S1) and signal amplified using a DakoCytomation-catalyzed system. Secondary antibodies were used as a starting point for amplification. Slides were scanned, analyzed, and quantified using Microvigene software (VigeneTech Inc., Carlisle, MA) to generate spot signal intensities, which were processed by the R package SuperCurve (version 1.01) [18 (link)], available at "http://bioinformatics.mdanderson.org/OOMPA". A fitted curve ("supercurve") was plotted with the signal intensities on the Y-axis and the relative log2 concentration of each protein on the X-axis using the non-parametric, monotone increasing B-spline model [18 (link)]. Protein concentrations were derived from the supercurve for each lysate by curve-fitting and normalized by median polish. Protein measurements were corrected for loading as described [15 (link)-17 (link),19 (link)]. For the selection of the 146 antibody set, we focused on markers currently used for breast cancer classification due to their value in treatment decisions (ER, PR, HER2). We then added additional antibodies to targets implicated in breast cancer pathophysiology, followed by antibodies to targets implicated in the pathophysiology of other cancer lineages. Final selection of antibodies was also driven by the availability of their high quality that could pass a strict validation process as previously described [20 (link)].
Gonzalez-Angulo A.M., Hennessy B.T., Meric-Bernstam F., Sahin A., Liu W., Ju Z., Carey M.S., Myhre S., Speers C., Deng L., Broaddus R., Lluch A., Aparicio S., Brown P., Pusztai L., Symmans W.F., Alsner J., Overgaard J., Borresen-Dale A.L., Hortobagyi G.N., Coombes K.R, & Mills G.B. (2011). Functional proteomics can define prognosis and predict pathologic complete response in patients with breast cancer. Clinical proteomics, 8(1), 11.
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Publication 2011
Antibodies Aspiration Biopsy, Fine-Needle bicinchoninic acid Biological Assay Buffers Epistropheus ERBB2 protein, human Freezing Homo sapiens Immunoglobulins Malignant Neoplasm of Breast Malignant Neoplasms Neoplasms Nitrocellulose Proteins Technique, Dilution
3
Tissue Homogenization and Protein Extraction
Cited 20 times

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Publication 2016
bicinchoninic acid Brain Buffers Cold Temperature Dithiothreitol Freezing Iodoacetamide lysyl endopeptidase Nucleic Acids Peptide Hydrolases Peptides Phosphoric Monoester Hydrolases Promega Proteins SDS-PAGE Sep-Pak C18 Stainless Steel Tissues Trypsin Urea Vacuum
4
SILAC-Based HeLa Cell Proteome Analysis
Cited 19 times
Hela cells were grown over 8 passages in High Glucose DMEM [12.43 g/l Dulbecco’s Modified Eagle’s Medium (Caisson Laboratories Inc., North Logan, UT, USA), 4.5 g/l D-(+)-Glucose anhydrous (Fluka, Buchs, Switzerland), 30 mg/l Glycine (Fluka)] supplemented with either light (Sigma-Aldrich) or heavy (13C 15N, Sigma-Aldrich) isotope-labeled lysine and arginine at 37°C and 5% CO2.
Cells were harvested at 80% confluency by trypsinization, washed three times with ice cold PBS (GIBCO (Invitrogen), Paisley, UK) and the cell number was determined using a Neubauer chamber. Hela cells were spun down at 300 ×g and resuspended in one cell pellet volume PBS. Two pellet volumes of 8 M Urea (Sigma-Aldrich, Buchs, Switzerland) containing 50 mM ammonium bicarbonate (Sigma-Aldrich) and 0.1% RapiGest (Waters, Baden, Switzerland) were thoroughly mixed with the resuspended cells. Subsequent to sonication (80% amplitude, 0.6 cycle, 1 min) cell debris was spun down at 16000×g.
The protein concentration of the lysate was measured by BCA assay (bicinchoninic acid, Thermo Scientific, Reinach, Switzerland). Proteins were reduced with 5 mM TCEP (tris(2-carboxyethyl)phosphine, Thermo Scientific) at 37°C for 15 min and alkylated with 10 mM iodoacetamide (Sigma-Aldrich) for 30 min in the dark. Proteins were first digested with lysyl endopeptidase (Wako Chemicals, Neuss, Germany) at an enzyme – substrate ratio of 1 to 50 (w/w) at 35°C for 2 hours. After dilution with 50 mM ammonium bicarbonate to 0.8M urea trypsin (Promega) was added at the same ratio. Tryptic digestion was carried out overnight at 37 °C. Peptides were acidified with 1% trifluoroacetic acid (TFA, Thermo Scientific) and purified by solid-phase extraction using C18 cartridges (Sep-Pak, Waters). The SPE eluate was evaporated to dryness and reconstituted in 3% acetonitrile (Thermo Scientific) and 0.2% formic acid (Sigma-Aldrich).
Escher C., Reiter L., MacLean B., Ossola R., Herzog F., Chilton J., MacCoss M.J, & Rinner O. (2012). Using iRT, a normalized retention time for more targeted measurement of peptides. Proteomics, 12(8), 1111-1121.
Publication 2012
acetonitrile ammonium bicarbonate Arginine bicinchoninic acid Biological Assay Cells Cold Temperature Digestion Eagle Enzymes formic acid Glucose Glycine HeLa Cells Iodoacetamide Isotopes Lysine lysyl endopeptidase Peptides phosphine Promega Proteins Solid Phase Extraction Technique, Dilution TNFSF14 protein, human Trifluoroacetic Acid tris(2-carboxyethyl)phosphine Tromethamine Trypsin Urea
5
Protein Quantification by BCA Assay
Cited 14 times
Concentration of protein extracts can be determined in a variety of ways. The most commonly used may be the Bicinchoninic Acid assays (BCA). BCA assays involve reduction of copper ions in a temperature dependant fashion with the level of reduction correlating with protein concentration. Reduced copper ions bind to BCA forming a purple product which can be detected at 562 nm. Each run includes a dilution series of a known protein standard – bovine serum albumin (BSA) as a reference curve to allow determination of absolute protein concentrations. As each reaction set can be subtly influenced by incubation time and temperature, samples which will be grouped together for analysis should routinely be assayed together against the same standard curve. Here we employed a series of protein dilutions (1, 5, 10, 20, 30, 40 µg) and (0.5, 1, 2, 4, 6, 8, 10, 12, 14 µg) which were produced from mouse whole brain homogenate. Preparation of the dilution series was carried out after the concentration of protein had been determined using a micro BCA assay (Pierce). Briefly, two 4–12% Bis-tris gels were loaded; one stained for total protein (Coomassie or Instant Blue) and the other was transferred for QWB as above. Visualisation and quantification was carried out using the Li-COR Odyssey imager and software. See above.
Eaton S.L., Roche S.L., Llavero Hurtado M., Oldknow K.J., Farquharson C., Gillingwater T.H, & Wishart T.M. (2013). Total Protein Analysis as a Reliable Loading Control for Quantitative Fluorescent Western Blotting. PLoS ONE, 8(8), e72457.
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Publication 2013
bicinchoninic acid Biological Assay Bistris Brain Copper Gels Ions Mice, House Proteins Serum Albumin, Bovine Staphylococcal Protein A Technique, Dilution

Most recents protocols related to «Bicinchoninic acid»

1
Protein Leakage Assessment After Disinfection
The bicinchoninic acid (BCA) assay was conducted to assess the amount of protein leakage after the disinfection treatment [18 (link)]. After undergoing UV irradiation, exposure to an electric field, and combined disinfection methods, the agar blocks were subjected to elution in 10 mL of PBS. Subsequently, 1 mL of the eluent was transferred into a tube and centrifuged at a speed of 8000 rpm for 5 min. The BCA assay was performed according to the protocol provided with the kit.
Zhang Y., Liang Y., Pan D., Bai S., Wen D., Tang M., Song H., Guo X, & Han H. (2024). Enhancing Escherichia coli Inactivation: Synergistic Mechanism of Ultraviolet Light and High-Voltage Electric Field. Foods, 13(9), 1343.
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Publication 2024
2
Protein Quantification with BCA Assay
To determine protein content the Pierce Bicinchoninic Acid (BCA) Protein Assay Kit (Thermo Fisher Scientific) was used following the manufacturer’s instructions.
de Zeeuw P., Treps L., García-Caballero M., Harjes U., Kalucka J., De Legher C., Brepoels K., Peeters K., Vinckier S., Souffreau J., Bouché A., Taverna F., Dehairs J., Talebi A., Ghesquière B., Swinnen J., Schoonjans L., Eelen G., Dewerchin M, & Carmeliet P. (2024). The gluconeogenesis enzyme PCK2 has a non-enzymatic role in proteostasis in endothelial cells. Communications Biology, 7, 618.
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Publication 2024
3
Protein Quantification via BCA Assay
A bicinchoninic acid (BCA) assay was performed for protein quantification according to manufacturer's instructions of the Micro BCA Protein Kit (Life Technologies, 23235). Absorbance was determined using a Multiskan™ FC Microplate Photometer (ThermoFisher Scientific). BCA is used for all quantification of sEVs for functional assays in this study.
van de Wakker S.I., Bauzá‐Martinez J., Ríos Arceo C., Manjikian H., Snijders Blok C.J., Roefs M.T., Willms E., Maas R.G., Pronker M.F., de Jong O.G., Wu W., Görgens A., El Andaloussi S., Sluijter J.P, & Vader P. (2024). Size matters: Functional differences of small extracellular vesicle subpopulations in cardiac repair responses. Journal of Extracellular Vesicles, 13(1), 12396.
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Publication 2024
4
Quantifying Exosome Protein Content
The total exosome content was extracted for quantification with a Bicinchoninic (BCA) Acid Protein Assay Kit (DNA Biotech, Iran), consisting of a standard solution, copper, and BCA reagents. The standard curve was plotted at different levels (50–250 µg/mL) against bovine serum albumin (BSA) as the standard solution. The exosomes and standard solutions were separately mixed with a mixture of copper and BCA reagents at a ratio of 1:50 and incubated at 60 °C for 60 min. Finally, the absorbance of the samples was read with a spectrophotometer at 562 nm (MPR4 + ; Hyperion, Roeder mark, Germany).
Nasiri Z., Soleimanjahi H., Baheiraei N., Hashemi S.M, & Pourkarim M.R. (2024). The impact understanding of exosome therapy in COVID-19 and preparations for the future approaches in dealing with infectious diseases and inflammation. Scientific Reports, 14, 5724.
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Publication 2024
5
Optimizing Soluble Protein Extraction from Snakehead Fish
Not available on PMC !
Total soluble protein was assayed using Pierce Bicinchoninic Acid (BCA) protein assay (ThermoFisher, USA) to establish the optimal approach for recovering the total amount of soluble protein from the flesh of snakehead fish. Its absorbance was measured at 540 nm in an ELISA plate reader. Bovine serum albumin (BSA) was used as the standard protein. The analysis was performed following the protocol provided by ThermoFisher Scientific. The ratio 1:8 of sample to working reagent was prepared, loaded into a 96-well plate, and incubated at 37 °C for 30 min. The total soluble protein concentration was quantified according to the standard curve obtained from the protein standard.
Setiawan H., Intanon M., CHUMMITRI P., Sringarm K., & Sathanawongs A. (2024). Snakehead Fish, Channa striata (Bloch, 1973), Protein Concentrate: Excellent Recovery of Fish-Based Albumin Source and Its Possible Application for Sperm Capacitation. Asian Fisheries Science, 37(2).
Publication 2024

Top products related to «Bicinchoninic acid»

1
Pvdf membrane by Merck Group
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
2
Bca protein assay kit by Thermo Fisher Scientific
Sourced in United States, China, Germany, United Kingdom, Japan, Belgium, France, Switzerland, Italy, Canada, Australia, Sweden, Spain, Israel, Lithuania, Netherlands, Denmark, Finland, India, Singapore
The BCA Protein Assay Kit is a colorimetric detection and quantification method for total protein concentration. It utilizes bicinchoninic acid (BCA) for the colorimetric detection and quantification of total protein. The assay is based on the reduction of Cu2+ to Cu1+ by protein in an alkaline medium, with the chelation of BCA with the Cu1+ ion resulting in a purple-colored reaction product that exhibits a strong absorbance at 562 nm, which is proportional to the amount of protein present in the sample.
3
Bicinchoninic acid protein assay kit by Thermo Fisher Scientific
Sourced in United States, China, Japan, Canada, France, United Kingdom, Macao
The Bicinchoninic acid (BCA) protein assay kit is a colorimetric detection and quantification method used to measure the total protein concentration in a sample. The kit utilizes the reduction of Cu2+ to Cu+ by protein in an alkaline medium, and the subsequent colorimetric detection of the Cu+ by bicinchoninic acid.
4
Bicinchoninic acid assay by Thermo Fisher Scientific
Sourced in United States, Germany, United Kingdom, China, New Zealand, Belgium, Denmark
The Bicinchoninic acid (BCA) assay is a colorimetric detection and quantification method used to measure the total protein concentration in a sample. The assay relies on the reduction of copper ions by proteins in an alkaline medium, which results in the formation of a purple-colored complex that can be detected spectrophotometrically.
5
Ripa lysis buffer by Beyotime
Sourced in China, United States, Switzerland, Germany, Japan, United Kingdom
RIPA lysis buffer is a detergent-based buffer solution designed for the extraction and solubilization of proteins from cells and tissues. It contains a mixture of ionic and non-ionic detergents that disrupt cell membranes and solubilize cellular proteins. The buffer also includes additional components that help to maintain the stability and activity of the extracted proteins.
6
Bca protein assay kit by Beyotime
Sourced in China, United States, Germany, Puerto Rico, United Kingdom, Switzerland, Japan, Sweden
The BCA protein assay kit is a colorimetric-based method for the quantitative determination of total protein concentration in a sample. It uses bicinchoninic acid (BCA) to detect and quantify the presence of protein.
7
Bicinchoninic acid protein assay kit by Beyotime
Sourced in China, United States
The Bicinchoninic acid (BCA) protein assay kit is a colorimetric detection and quantification method for proteins. The kit utilizes the reduction of Cu2+ to Cu1+ by proteins in an alkaline medium, followed by the highly sensitive and selective colorimetric detection of the Cu1+ cation using a reagent containing bicinchoninic acid.
8
Polyvinylidene fluoride membrane by Merck Group
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Polyvinylidene fluoride (PVDF) membranes are a type of lab equipment used for a variety of filtration and separation applications. They are made from a thermoplastic fluoropolymer material and offer high chemical and thermal resistance. PVDF membranes are commonly used in processes such as microfiltration, ultrafiltration, and sample preparation.
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Polyvinylidene difluoride membrane by Merck Group
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Polyvinylidene difluoride (PVDF) membranes are a type of lab equipment used for various applications. PVDF membranes are known for their chemical resistance, thermal stability, and mechanical strength. They are commonly used in filtration, separation, and transfer processes in laboratory settings.
10
Protease inhibitor cocktail by Merck Group
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The Protease Inhibitor Cocktail is a laboratory product designed to inhibit the activity of proteases, which are enzymes that can degrade proteins. It is a combination of various chemical compounds that work to prevent the breakdown of proteins in biological samples, allowing for more accurate analysis and preservation of protein integrity.

More about "Bicinchoninic acid"

Bicinchoninic acid (BCA) is a chemical compound widely used in colorimetric assays to quantify the total protein concentration in a sample.
It works by chelating copper ions, which are then reduced by the proteins present, resulting in a purple-colored complex that can be measured spectrophotometrically.
The BCA assay is a sensitive, reproducible, and widely-used method for protein quantification in biochemistry and cell biology research.
Optimizing the BCA assay protocol can boost research productivity and reproducibility.
PubCompare.ai's AI-driven platform can help researchers identify the most effective and reproducible BCA assay methods from literature, preprints, and patents.
This can be particularly useful when working with PVDF membranes, RIPA lysis buffer, and protease inhibitor cocktails, which are commonly used in protein analysis workflows.
The BCA protein assay kit is a popular commercial option for quantifying protein samples.
It utilizes the same bicinchoninic acid principle as the BCA assay, providing a convenient and reliable way to measure protein concentrations.
By leveraging AI-powered comparisons of available BCA assay protocols, researchers can optimize their protein quantification methods and improve the reproducibility of their experiments, ultimately enhancing the quality and impact of their research.