Estimating Protein Concentration by Dot Blotting of Multiple Samples

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4253
Keyword(s):  
Protein Concentration ◽  
Multiple Samples ◽  
Dot Blotting

scholarly journals Standard Flow Multiplexed Proteomics (SFloMPro)—An Accessible Alternative to NanoFlow Based Shotgun Proteomics

Proteomes ◽  
2022 ◽  
Vol 10 (1) ◽  
pp. 3
Author(s):  
Benjamin C. Orsburn ◽  
Sierra D. Miller ◽  
Conor J. Jenkins
Keyword(s):  
Protein Concentration ◽  
Shotgun Proteomics ◽  
High Coverage ◽  
Mass Spectrometers ◽  
Chemical Tag ◽  
Peptide Labeling ◽  
Isobaric Tagging ◽  
Pump Pressure ◽  
Multiple Samples ◽  
Chemical Label

Multiplexed proteomics using isobaric tagging allows for simultaneously comparing the proteomes of multiple samples. In this technique, digested peptides from each sample are labeled with a chemical tag prior to pooling sample for LC-MS/MS with nanoflow chromatography (NanoLC). The isobaric nature of the tag prevents deconvolution of samples until fragmentation liberates the isotopically labeled reporter ions. To ensure efficient peptide labeling, large concentrations of labeling reagents are included in the reagent kits to allow scientists to use high ratios of chemical label per peptide. The increasing speed and sensitivity of mass spectrometers has reduced the peptide concentration required for analysis, leading to most of the label or labeled sample to be discarded. In conjunction, improvements in the speed of sample loading, reliable pump pressure, and stable gradient construction of analytical flow HPLCs has continued to improve the sample delivery process to the mass spectrometer. In this study we describe a method for performing multiplexed proteomics without the use of NanoLC by using offline fractionation of labeled peptides followed by rapid “standard flow” HPLC gradient LC-MS/MS. Standard Flow Multiplexed Proteomics (SFloMPro) enables high coverage quantitative proteomics of up to 16 mammalian samples in about 24 h. In this study, we compare NanoLC and SFloMPro analysis of fractionated samples. Our results demonstrate that comparable data is obtained by injecting 20 µg of labeled peptides per fraction with SFloMPro, compared to 1 µg per fraction with NanoLC. We conclude that, for experiments where protein concentration is not strictly limited, SFloMPro is a competitive approach to traditional NanoLC workflows with improved up-time, reliability and at a lower relative cost per sample.

Albumin-Induced Endocytic Vacuoles in Tetrahymena Pyrijormis

1975 ◽  
Vol 33 ◽  
pp. 694-695
Author(s):  
L. J. Brenner ◽  
D. G. Osborne ◽  
B. L. Schumaker
Keyword(s):  
Electron Microscopy ◽  
Bovine Serum Albumin ◽  
Bovine Serum ◽  
Protein Concentration ◽  
Tetrahymena Pyriformis ◽  
Sequence Of Events ◽  
Cytoplasmic Bodies ◽  
Food Vacuoles ◽  
Mouse Ascites ◽  
Normal Human

Exposure of the ciliate, Tetrahymena pyriformis, strain WH6, to normal human or rabbit sera or mouse ascites fluids induces the formation of large cytoplasmic bodies. By electron microscopy these (LB) are observed to be membrane-bounded structures, generally spherical and varying in size (Fig. 1), which do not resemble the food vacuoles of cells grown in proteinaceous broth. The possibility exists that the large bodies represent endocytic vacuoles containing material concentrated from the highly nutritive proteins and lipoproteins of the sera or ascites fluids. Tetrahymena mixed with bovine serum albumin or ovalbumin solutions having about the same protein concentration (7g/100 ml) as serum form endocytic vacuoles which bear little resemblance to the serum-induced LB. The albumin-induced structures (Fig. 2) are irregular in shape, rarely spherical, and have contents which vary in density and consistency. In this paper an attempt is made to formulate the sequence of events which might occur in the formation of the albumin-induced vacuoles.

Short Stress State Questionnaire

2015 ◽  
Vol 31 (1) ◽  
pp. 20-30 ◽  
Author(s):  
William S. Helton ◽  
Katharina Näswall
Keyword(s):  
Stress State ◽  
Factor Structure ◽  
Human Performance ◽  
Self Report ◽  
Confirmatory Factor ◽  
Stress States ◽  
Related Stress ◽  
Using Data ◽  
Task Conditions ◽  
Multiple Samples

Conscious appraisals of stress, or stress states, are an important aspect of human performance. This article presents evidence supporting the validity and measurement characteristics of a short multidimensional self-report measure of stress state, the Short Stress State Questionnaire (SSSQ; Helton, 2004 ). The SSSQ measures task engagement, distress, and worry. A confirmatory factor analysis of the SSSQ using data pooled from multiple samples suggests the SSSQ does have a three factor structure and post-task changes are not due to changes in factor structure, but to mean level changes (state changes). In addition, the SSSQ demonstrates sensitivity to task stressors in line with hypotheses. Different task conditions elicited unique patterns of stress state on the three factors of the SSSQ in line with prior predictions. The 24-item SSSQ is a valid measure of stress state which may be useful to researchers interested in conscious appraisals of task-related stress.

Anxious attachment predicts rejection sensitivity across multiple samples

2011 ◽  
Author(s):  
Kenzie Snyder ◽  
Gertraud Stadler ◽  
M. Joy McClure ◽  
Niall Bolger
Keyword(s):  
Rejection Sensitivity ◽  
Anxious Attachment ◽  
Multiple Samples

Monoamine Oxidase and Other Mitochondrial Enzymes in Density Subpopulations of Human Platelets

1988 ◽  
Vol 59 (01) ◽  
pp. 029-033 ◽  
Author(s):  
K G Chamberlain ◽  
D G Penington
Keyword(s):  
Monoamine Oxidase ◽  
Protein Concentration ◽  
Close Correlation ◽  
Human Platelets ◽  
Mitochondrial Enzymes ◽  
Density Fractions ◽  
Percoll Gradients ◽  
Normal Human ◽  
The Mean ◽  
Very High

SummaryNormal human platelets have been separated according to density on continuous Percoll gradients and the platelet distribution divided into five fractions containing approximately equal numbers of platelets. The mean volumes and protein contents of the platelets in each fraction were found to correlate positively with density while the protein concentration did not differ significantly between the fractions. Four mitochondrial enzymes (monoamine oxidase, glutamate dehydrogenase, cytochrome oxidase and NADP-dependent isocitrate dehydrogenase) were assayed and their activities per unit volume were found to increase in a very similar monotonie fashion with platelet density. When MAO and GDH were assayed on the same set of density fractions the correlation between the two activities was very high (r = 0.94–1.00, p <0.001) and a similar close correlation was found between MAO and ICDH. The results support the hypothesis that high density platelets either have a higher concentration of mitochondria or have larger mitochondria than low density platelets.

Does Low Protein Concentration of Tissue-type Plasminogen Activator Predict a Low Risk of Spontaneous Deep Vein Thrombosis?

1995 ◽  
Vol 74 (02) ◽  
pp. 718-721 ◽  
Author(s):  
Jørgen Gram ◽  
Johannes Sidelmann ◽  
Jørgen Jespersen
Keyword(s):  
Deep Vein Thrombosis ◽  
Confidence Interval ◽  
Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Protein Concentration ◽  
Tissue Type ◽  
Vein Thrombosis ◽  
Low Protein ◽  
Deep Vein ◽  
Pai 1

SummaryMany reports have demonstrated an abnormal fibrinolysis in a subset of patients with deep vein thrombosis. We have studied systemic global fibrinolytic activity and protein concentrations of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor type 1 (PAI-1) in plasma of 25 young patients with a previous instance of spontaneous deep vein thrombosis documented by phlebography and in 50 healthy controls. The two populations were comparable with respect to a number of base-line variables (age, height, weight, etc.), while the patients had significantly lower fibrinolytic activity (p <0.02), and significantly higher protein concentrations of t-PA (p <0.0001) and PAI-1 (p <0.0006).We used probit scale plots to identify the consequence of different cut-off points to separate patients from controls. Reasonable separation could be obtained for t-PA with a cut-off point of 5.2 ng/ml and for PAI-1 18 ng/ml. The sensitivity and specificity for these cut-off points were for t-PA 73% (95% confidence interval 63%-84%) and for PAI-1 67% (confidence interval 55%-77%). The negative predictive value with a cut-off point t-PA concentration of 5.2 ng/ml was 85% (95% confidence interval 70%-94%). We observed a significantly negative association between concentration of t-PA and fibrinolytic activity (rs = -0.47; p <0.005) and also between PAI-1 and fibrinolytic activity (rs = -0.78; p <0.005).We conclude that a young healthy population is characterized by low protein concentration of t-PA (and PAI-1) compared with young patients with a previous instance of spontaneous vein thrombosis, and we tentatively state that a low protein concentration of t-PA predicts a low risk of spontaneous deep vein thrombosis.

Extraction and Partial Purification of Protease from Bilimbi (Averrhoa bilimbi L.)

2013 ◽  
Vol 10 (2) ◽  
pp. 29
Author(s):  
Normah Ismail ◽  
Nur' Ain Mohamad Kharoe
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
Ammonium Sulfate ◽  
Molecular Weight Distribution ◽  
Weight Distribution ◽  
Protease Activity ◽  
Protein Concentration ◽  
Temperature Stability ◽  
Partial Purification ◽  
Ammonium Sulfate Precipitation

Unripe and ripe bilimbi (Averrhoa bilimbi L.) were ground and the extracted juices were partially purified by ammonium sulfate precipitation at the concentrations of 40 and 60% (w/v). The collected proteases were analysed for pH, temperature stability, storage stability, molecular weight distribution, protein concentration and protein content. Protein content of bilimbi fruit was 0.89 g. Protease activity of both the unripe and ripe fruit were optimum at pH 4 and 40°C when the juice were purified at 40 and 60% ammonium sulfate precipitation. A decreased in protease activity was observed during the seven days of storage at 4°C. Molecular weight distribution indicated that the proteases protein bands fall between IO to 220 kDa. Protein bands were observed at 25, 50 and 160 kDa in both the unripe and ripe bilimbi proteases purified with 40% ammonium sulfate, however, the bands were more intense in those from unripe bilimbi. No protein bands were seen in proteases purified with 60% ammonium sulfate. Protein concentration was higher for proteases extracted with 40% ammonium sulfate at both ripening stages. Thus, purification using 40% ammonium sulfate precipitation could be a successful method to partially purify proteases from bilimbi especially from the unripe stage. 

Formulation of Colors Using a Genetic Algorithm

2012 ◽  
Vol 17 (4) ◽  
pp. 241-244
Author(s):  
Cezary Draus ◽  
Grzegorz Nowak ◽  
Maciej Nowak ◽  
Marcin Tokarski
Keyword(s):  
Genetic Algorithm ◽  
Genetic Algorithms ◽  
Production Process ◽  
Computer Technology ◽  
Manual Method ◽  
Apply Genetic ◽  
Computer Aided ◽  
Multiple Samples ◽  
Plastics Industry

Abstract The possibility to obtain a desired color of the product and to ensure its repeatability in the production process is highly desired in many industries such as printing, automobile, dyeing, textile, cosmetics or plastics industry. So far, most companies have traditionally used the "manual" method, relying on intuition and experience of a colorist. However, the manual preparation of multiple samples and their correction can be very time consuming and expensive. The computer technology has allowed the development of software to support the process of matching colors. Nowadays, formulation of colors is done with appropriate equipment (colorimeters, spectrophotometers, computers) and dedicated software. Computer-aided formulation is much faster and cheaper than manual formulation, because fewer corrective iterations have to be carried out, to achieve the desired result. Moreover, the colors are analyzed with regard to the metamerism, and the best recipe can be chosen, according to the specific criteria (price, quantity, availability). Optimaization problem of color formulation can be solved in many diferent ways. Authors decided to apply genetic algorithms in this domain.

Sign in / Sign up

Export Citation Format

Share Document