Analytical Techniques in Biochemistry and Molecular Biology

Katoch, Rajan

doi:10.1007/978-1-4419-9785-2

Cited by 41 publications

(31 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The reagent which is initially yellow reacts with free alpha amino groups present in all amino acids, proteins, or peptides and forms a deep blue or purple colored complex known as Ruhemann's purple. Its absorb an ceis measured at 570 nm using aspectrophotometer [26].…”

Section: Ninhydrin Testmentioning

confidence: 99%

Musculoskeletal ProteinAnalysis Techniques - A Review

Kamineni¹,

Manepally²,

Kamineni³

2016

JRAD

View full text Add to dashboard Cite

There have been a large number of experimental methods for purifying and analyzing proteins from the sample of interest. Determination of protein concentration is often the key step and is common to many applications in protein research and life sciences. The most commonly used quantitative techniques to measure the protein concentration are Amino acid analysis, Biuret assay, Bradford assays, Folin-Lowry assay, Bicinchoninic (BCA) assays, UV absorbance assays and Antibody-based assays such as ELISA and Western Blotting. Some of the qualitative methods that are used to detect different types of proteins and amino acids are Ninhydrin test, Xanthoproteic test, Millon's test, Sulfur test, Hopkins-Cole test, and Nitroprusside test. Chromatographic analysis can also be carried out on an either qualitative or quantitative basis to purify complex protein mixtures based on their properties such as size, solubility, charge, hydrophobicity and bio-specific interaction. Selection of these assays is based upon the ease of performance, range of concentrations, sensitivity, and interfering substances contained in the complex mixture. The purpose of this review is to provide an assessment of commonly used chromatographic and colorimetric methods for the determination of protein concentration.

show abstract

Section: Ninhydrin Testmentioning

confidence: 99%

Musculoskeletal ProteinAnalysis Techniques - A Review

Kamineni¹,

Manepally²,

Kamineni³

2016

JRAD

View full text Add to dashboard Cite

show abstract

“…The extent of proteins hydrolyzed was determined by measuring the free amino acid (FFA) content of hydrolysates. Ninhydrin, a powerful oxidizing agent, decarboxylates alpha-amino acids and yields a bluish purple product measured at 570 nm (Katoch, 2011). FFA content of hydrolysates is a good index of their degree of the hydrolysis of proteins.…”

Section: Results and Discussion 241 Determination Of Protein And Frmentioning

confidence: 99%

Inhibitory Effects of Hydrolyzed Oat Bran Proteins on Human LDL Oxidation and Their Bile Acid Binding Capacity

Espinosa¹,

Yunuen²

View full text Add to dashboard Cite

Free radicals attack to biological molecules can initiate their oxidation and contribute to the development of several chronic diseases. Oxidation of low density lipoprotein, as well as a higher concentration of cholesterol, have been implicated in the development of atherosclerotic lesions and increased risks of cardiovascular diseases. Strategies to decrease the risks then include scavenging of free radicals and the reduction of cholesterol (e.g. binding of bile acids). In this regard, the aim of this project was; 1) to produce and characterize various hydrolyzed proteins from oat brans; 2) determine their antioxidant, metal binding and bile chelating capacities as well as their ability to inhibit low density lipoprotein oxidation (LDL). Medium oat bran samples were treated with two cell wall polysaccharide degrading enzymes (cellulase and viscozyme) to generate two protein isolates named CPI and VPI respectively. Ten hydrolysates were subsequently prepared by treating CPI or VPI with five proteases protamex, alcalase, flavourzyme, pepsin and pepsin+pancreatin. The two highest degrees of hydrolysis (26.4 ± 0.5 and 22.0 ± 0.1%) were obtained under simulated gastrointestinal digestion with pepsin and pancreatin. Intrinsic absorbance showed that CPI-alcalase and CPI-pepsin/pancreatin had a higher content of aromatic amino acids. In the antioxidant oxidant assays, VPI-pepsin better scavenged ROO • radicals (496.77±5.83 μM Trolox Equivalents (TE)/g) while VPI-flavourzyme and VPI-pepsin had better quenching for HO • (27.95 ±1.580 and and O2 •-(45.31±6.6%) radicals, respectively. In the metal

show abstract

“…Often, a series of purification steps are executed to ensure homogeneity of the final product. 53 Ion exchange chromatography is one purification method that can be employed. Ion exchange chromatography separates proteins based on charge type and relative strength; it is dependent on the interaction between charged amino acid residues in the sample and ions in the column stationary phase.…”

Section: Protein Purification Techniquesmentioning

confidence: 99%

“…59 Larger proteins that cannot move through the pores instead travel around the matrix and elute prior to smaller proteins; because small proteins can fit through all of the pores in the matrix, they take a longer path and thus elute last. 53 Because purification is not dependent on binding of the target protein, this method is generally less susceptible to protein loss. 53…”

Section: Protein Purification Techniquesmentioning

confidence: 99%

See 1 more Smart Citation

Recombinant Expression and Potential Autocatalysis of Aedes aegypti Trypsin-like Serine Proteases (AaSPII and AaSPIV)

Eilerts

View full text Add to dashboard Cite

Aedes aegypti mosquitoes can be found globally in tropical and subtropical urban areas and spread Zika, Dengue fever, yellow fever, and Chikungunya viruses. Current vector control methods are limited and nonspecific. The female Ae. aegypti mosquito uses blood meal proteins to obtain nutrients required for oogenesis; inhibition of the midgut trypsin-like serine proteases responsible for blood meal digestion may provide a novel method of vector control. Ae. aegypti blood meal digestion is complex and the role of uncharacterized serine proteases in blood digestion is unclear; specifically, a group of trypsin-like serine proteases (AaSPII-V) is expressed at constant levels before and following Ae. aegypti blood meal acquisition. This research focuses on the in vitro biochemical study of two specific Ae. aegypti trypsin-like serine proteases (AaSPII and AaSPIV) in order to gain further understanding of their role in blood meal digestion. The approach involved the successful cloning and bacterial expression of these soluble, recombinant proteases. Results from attempts to purify these proteases were unsuccessful but indicative of potential autocatalytic and autodigestive behavior. Future studies will focus on obtaining purified recombinant proteases for further study. The study of AaSPII and AaSPIV, as well as other midgut Ae. aegypti proteases, will aid in understanding the overall role proteases play in blood meal digestion and may eventually allow for the development of mosquito-specific enzyme inhibitors. v ACKNOWLEDGMENTS First and foremost, I would like to thank Prof. Alberto A. Rascón Jr., my research advisor, for the opportunity to perform this research in his laboratory and for the expert mentorship and guidance I received throughout my research endeavors. While working with Prof. Rascón, I gained invaluable experience and learned both technical and practical skills to help me succeed in the future as a research scientist. I would also like to extend my gratitude to the members of my graduate committee, Prof. Laura Miller Conrad and Prof. Marc d'Alarcao, for their support and assistance in developing this thesis. I also thank my friends and colleagues at San Jose State University and in the Rascón Lab for the scientific discourse, encouragement, and laughter we've shared. For their constant love and support, I want to thank my family and friends. My mother and father, Valerie and Gregg Eilerts, have provided a lifetime of support and encouragement and prepared me for a successful future. I thank my brother, Tom Eilerts, for always making me laugh and for his endearing attempts to help me overcome research struggles via jelly donut metaphors. I thank my sister, Renee Eilerts, for always welcoming me in her home and making sure I remembered to occasionally have fun outside of the lab (Go Sharks!). I thank my best friend, Chelsea Carman, for listening and supporting me through both my undergraduate and graduate education. Lastly, I want to extend my deepest gratitude to my partner, Morgan Hoffman, for his unwaver...

show abstract

Analytical Techniques in Biochemistry and Molecular Biology

Cited by 41 publications

References 4 publications

Musculoskeletal ProteinAnalysis Techniques - A Review

Musculoskeletal ProteinAnalysis Techniques - A Review

Inhibitory Effects of Hydrolyzed Oat Bran Proteins on Human LDL Oxidation and Their Bile Acid Binding Capacity

Recombinant Expression and Potential Autocatalysis of Aedes aegypti Trypsin-like Serine Proteases (AaSPII and AaSPIV)

Contact Info

Product

Resources

About