Interpretation of protein quantitation using the Bradford assay: Comparison with two calculation models

Ku, Hyung-Keun; Lim, Hyuk-Min; Oh, Kyong-Hwa; Yang, Hyo-Jin; Jeong, Ji‐Seon; Kim, Sook-Kyung

doi:10.1016/j.ab.2012.10.045

Cited by 55 publications

(23 citation statements)

References 8 publications

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“…Lysozyme (10 mg/ mL) was incubated with 0.5 M ribose in 0.1 M sodium phosphate buffer containing 3 mM sodium azide, pH 7.4 at 37 C for 24 h. Unbound ribose was removed by dialysis against 4 L of 0.1 M sodium phosphate buffer, pH 7.4 at 4 C for 48 h with 5-6 changes. Following dialysis, the protein concentration was determined using the Bio-Rad standard protein assay kit (Bio-Rad, Berkeley, CA) based on the Bradford dye-binding procedure (Ku et al, 2013). Dialyzed ribated lysozyme (10 mg/mL) was reincubated with 5-10 mg/ mL of AI and aminoguanidine in 0.1 M sodium phosphate buffer containing 3 mM sodium azide, pH 7.4 at 37 C for 15 d.…”

Section: Amadorin Activitymentioning

confidence: 99%

Beneficial effect ofAzadirachta indicaon advanced glycation end-product in streptozotocin-diabetic rat

Gutierrez

Martı́nez-Ortiz

2014

Pharmaceutical Biology

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Section: Amadorin Activitymentioning

confidence: 99%

Beneficial effect ofAzadirachta indicaon advanced glycation end-product in streptozotocin-diabetic rat

Gutierrez

Martı́nez-Ortiz

2014

Pharmaceutical Biology

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“…This indicated an effective action of the The decrease in protein concentration during in vitro digestion has also been reported in a soy milk bioaccessibility study and was attributed to the enzymatic hydrolysis of the protein-structure-forming peptides [26], which could not be detected by the Bradford protein assay. This is because the blue dye Coomassie Brilliant Blue G-250 (CBBG) has a low affinity for low-molecular-weight peptides (<3 kDa) [27][28][29][30].…”

Section: Protein Concentration and Degree Of Hydrolysis Of Microcapsumentioning

confidence: 99%

In Vitro Digestion of Microcapsule Carriers for Oral Delivery of Bioactive Compounds for Diabetes Treatment and Their Inhibitory Effect on the DPP-4 Enzyme

et al. 2019

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Empty microcapsules, originally designed as carriers of bioactive peptides, were prepared by the combined method of a double-emulsion complex with coacervation spray drying and were subjected to an in-vitro digestion process, producing peptides from the whey protein contained in the microcapsule walls. The inhibitory effect of the enzyme dipeptidyl peptidase-4 (DPP-4) and modulation of the insulin receptor of hydrolyzed microcapsules were evaluated. The hydrolysate of the microcapsules was subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis, showing the presence of low-molecular-weight peptidic compounds, which apparently were responsible for the DPP-4 inhibitory effect. Fluorescence analysis showed that the effect of the hydrolyzed microcapsules on the insulin receptor was 40% that of insulin. The inhibition of DPP-4 was 54.7%. This work demonstrated that empty microcapsules initially designed as carriers of functional peptides also have the capability to inhibit DPP-4 and modulate insulin receptors by themselves.

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“…After treatment, the cells were collected for 2 experiments: 1) Total RNA was extracted from the cells using TRizol reagent (Invitrogen) as described by Hong et al (2012) and (2) the cells were solubilized in Cell Extraction Buffer (Invitrogen) for 30 min and the lysates were cleared by centrifugation at 13,000 × g for 10 min at 4°C. The protein concentration of each sample was quantified by the Bradford method (Ku et al, 2013).…”

Section: Invitrobiologicalfunctionanalysismentioning

confidence: 99%

Characterization and functional analyses of a novel chicken CD8α variant X1 (CD8α1)1,2

Truong

Ban

Park

et al. 2016

Journal of Animal Science

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We provide the first description of cloning and of structural and functional analysis of a novel variant in the chicken cluster of differentiation 8 alpha (CD8a) family, termed the CD8α X1 (CD8α1) gene. Multiple alignments of CD8α1 with known CD8α and CD8β sequences of other species revealed relatively low conservation of AA residues involved in the specific and unique structural domains among CD8α genes. For example, cysteine residues that are involved in disulfide bonding to form the V domain are conserved. In contrast, the O-linked glycosylation sites (XPXX motif) are not found in the chicken CD8α1 sequence, and the A β strand and complementarity-determining region 1 and 2 sequences are poorly conserved between chicken CD8α1 and avian CD8α. Furthermore, the alignment showed that the transmembrane regions show relatively high sequence similarity, whereas the cytoplasmic regions show relatively low similarity, indicating poor conservation. Moreover, the motif (CXCP) that is thought to be responsible for binding the p56 lymphocyte cell kinase subunit (p56) is missing in the CD8α1 sequence. The chicken CD8α1 genomic structure is similar to that of chicken CD8α, but their protein structures differ. Phylogenetic analysis showed that chicken CD8α1 grouped with known avian CD8α sequences but was somewhat distantly related to the CD8α molecules of other species. Moreover, we analyzed the signal transduction and cytokine response to CD8α1 treatment to determine the specific biological functions of chicken CD8α1 in immune cells. The results showed that chicken CD8α1 is a key regulator of the expression of genes that are associated and cooperate with transcription factors in the major histocompatibility complex class I and II promoter regions and activates Janus kinase (JAK) 1/2, signal transducer and activator of transcription (STAT), and suppressor of cytokine signaling (SOCS) 1 signaling-related genes. Immune cells that express functional CD8α1 induce proinflammatory cytokines as well as innate immune responses. Therefore, our data indicate that CD8α1 may have immunoregulatory activity by regulating the expression of proinflammatory or anti-inflammatory cytokines via its effect on immune cells.

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Interpretation of protein quantitation using the Bradford assay: Comparison with two calculation models

Cited by 55 publications

References 8 publications

Beneficial effect ofAzadirachta indicaon advanced glycation end-product in streptozotocin-diabetic rat

Beneficial effect ofAzadirachta indicaon advanced glycation end-product in streptozotocin-diabetic rat

In Vitro Digestion of Microcapsule Carriers for Oral Delivery of Bioactive Compounds for Diabetes Treatment and Their Inhibitory Effect on the DPP-4 Enzyme

Characterization and functional analyses of a novel chicken CD8α variant X1 (CD8α1)1,2

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