Name: utilizing the ethylene-releasing compound, 2-chloroethylphosphonic acid, as a tool to study ethylene response in bacteria
Uploaded: 2016-11-10T14:00:00
Description: Watch this Scientific Journal Video about Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria at JoVE.com

The authors thank Dr. Dario Bonetta for providing Arabidopsis thaliana seeds and for technical assistance in regards to the triple response assay, as well as Simone Quaranta for help with FT-IR. This work was supported by a Natural Sciences and Engineering Research Council of Canada Discovery Grant (NSERC-DG) to JLS, an Ontario Graduate Scholarship (OGS) to RVA, and a Queen Elizabeth II Graduate Scholarship in Science and Technology (QEII-GSST) to AJV.

1. Chemicals
<ol>
	<li>Prepare a solution of 500 mM CEPA (144.49 g/mol), and a solution consisting of both 500 mM NaCl (58.44 g/mol) and 500 mM NaH2PO4&#183;H2O (137.99 g/mol) in acidified (pH 2.5) ultra-pure water or 0.1 N HCl. Mix using a vortex until the solutions are clear.</li>
	<li>Serially dilute (10x) the 500 mM solutions in the same solvent to obtain 5 mM and 50 mM stocks.</li>
	<li>Prepare a 10 mM solution of 1-aminocyclopropane carboxylic acid (ACC; 101.1 g/mol) in ultra-pure...

<ol>
	<li>Guzm&#225;n, P., Ecker, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploiting+the+triple+response+of+Arabidopsis+to+identify+ethylene-related+mutants.">Exploiting the triple response of Arabidopsis to identify ethylene-related mutants.</a> Plant Cell. 2 (6), 513-523 (1990).</li><li>Bakshi, A., Shemansky, J. M., Chang, C., Binder, B. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=History+of+research+on+the+plant+hormone+ethylene.">History of research on the plant hormone ethylene.</a> J. Plant Growth Regul. 34 (4), 809-827 (2015).</li><li>Schaller, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethylene+and+the+regulation+of+plant+development.">Ethylene and the regulation of plant development.</a> BMC Biol. 10 (1), (2012).</li><li>Hua, J., Sakai, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=EIN4+and+ERS2+are+members+of+the+putative+ethylene+receptor+gene+family+in+Arabidopsis.">EIN4 and ERS2 are members of the putative ethylene receptor gene family in Arabidopsis.</a> Plant Cell. 10 (8), 1321-1332 (1998).</li><li>Bleecker, A. B., Estelle, M. A., Somerville, C., Kende, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insensitivity+to+ethylene+conferred+by+a+dominant+Mutation+in+Arabidopsis+thaliana.">Insensitivity to ethylene conferred by a dominant Mutation in Arabidopsis thaliana.</a> Science. 241 (4869), 1086-1089 (1988).</li><li>Hamilton, A. J., Bouzayen, M., Grierson, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+a+tomato+gene+for+the+ethylene-forming+enzyme+by+expression+in+yeast.">Identification of a tomato gene for the ethylene-forming enzyme by expression in yeast.</a> Proc. Natl. Acad. Sci. 88 (16), 7434-7437 (1991).</li><li>Kim, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+ethylene+removal+with+Pseudomonas+strains.">Assessment of ethylene removal with Pseudomonas strains.</a> J. Hazard. Mater. 131 (3), 131-136 (2006).</li><li>Kim, H. E., Shitashiro, M., Kuroda, A., Takiguchi, N., Kato, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethylene+chemotaxis+in+Pseudomonas+aeruginosa+and+other+Pseudomonas+species.">Ethylene chemotaxis in Pseudomonas aeruginosa and other Pseudomonas species.</a> Microbes Environ. 22 (2), 186-189 (2007).</li><li>Augimeri, R. V., Strap, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+phytohormone+ethylene+enhances+bacterial+cellulose+production,+regulates+CRP/FNRKx+transcription+and+causes+differential+gene+expression+within+the+cellulose+synthesis+operon+of+Komagataeibacter+(Gluconacetobacter)+xylinus+ATCC+53582.">The phytohormone ethylene enhances bacterial cellulose production, regulates CRP/FNRKx transcription and causes differential gene expression within the cellulose synthesis operon of Komagataeibacter (Gluconacetobacter) xylinus ATCC 53582.</a> Front. Microbiol. 6, 1459 (2015).</li><li>Zhang, W., Wen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+ethylene+gas+and+comparison+of+ethylene+responses+induced+by+ethylene,+ACC,+and+ethephon.">Preparation of ethylene gas and comparison of ethylene responses induced by ethylene, ACC, and ethephon.</a> Plant Physiol. Biochem. 48 (1), 45-53 (2010).</li><li>Zhang, W., Hu, W., Wen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethylene+preparation+and+its+application+to+physiological+experiments.">Ethylene preparation and its application to physiological experiments.</a> Plant Signal. Behav. 5 (4), 453-457 (2010).</li><li>Warner, H. L., Leopold, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ethylene+evolution+from+2-chloroethylphosphonic+acid.">Ethylene evolution from 2-chloroethylphosphonic acid.</a> Plant Physiol. 44 (1), 156-158 (1969).</li><li>Biddle, E., Kerfoot, D. G. S., Kho, Y. H., Russell, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetic+studies+of+the+thermal+decomposition+of+2-chloroethylphosphonic+acid+in+aqueous+solution.">Kinetic studies of the thermal decomposition of 2-chloroethylphosphonic acid in aqueous solution.</a> Plant Physiol. 58 (5), 700-702 (1976).</li><li>Klein, I., Lavee, S., Ben-Tal, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+water+vapor+pressure+on+the+thermal+decomposition+of+2-chloroethylphosphonic+acid.">Effect of water vapor pressure on the thermal decomposition of 2-chloroethylphosphonic acid.</a> Plant Physiol. 63 (3), 474-477 (1979).</li><li>Murashige, T., Skoog, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+revised+medium+for+rapid+growth+and+bio+assays+with+tobacco+tissue+cultures.">A revised medium for rapid growth and bio assays with tobacco tissue cultures.</a> Physiol. Plant. 15 (3), 473-497 (1962).</li><li>Schramm, M., Hestrin, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+production+of+cellulose+at+the+air/liquid+interface+of+a+culture+of+Acetobacter+xylinum.">Factors affecting production of cellulose at the air/liquid interface of a culture of Acetobacter xylinum.</a> J. Gen. Microbiol. 11 (1), 123-129 (1954).</li><li>Schneider, C. A., Rasband, W. S., Eliceiri, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NIH+Image+to+ImageJ:+25+years+of+image+analysis.">NIH Image to ImageJ: 25 years of image analysis.</a> Nat. Methods. 9 (7), 671-675 (2012).</li><li>Ciolacu, D., Ciolacu, F., Popa, V. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amorphous+cellulose-structure+and+characterization.">Amorphous cellulose-structure and characterization.</a> Cellul. Chem. Technol. 45 (1), 13-21 (2011).</li></ol>

The methods described here outline the in situ production of ethylene from CEPA for the study of bacterial ethylene response using the model organism, K. xylinus. This method is very useful as ethylene can be produced by supplementing any aqueous medium that has a pH greater than 3.510,11 with CEPA negating the need for pure ethylene gas or specialized laboratory equipment. This method is not limited to studying the effects of CEPA-derived ethylene on bacteria but can be also be adapted to st...

The olefin ethylene (C2H4) was first discovered as a plant hormone in 1901 when it was observed that pea seedlings, grown in a laboratory that used coal gas lamps, exhibited an abnormal morphology in which stems (hypocotyls) were shorter, thicker and bent sideways compared to normal pea seedlings; a phenotype later termed the triple response1,2. Subsequent studies demonstrated that ethylene is a vital phytohormone that regulates numerous developmental processes such as growth, stress response, fruit ripening and senescence3. Arabidopsis thaliana, a model organism for plant biology research, has been well studied in regards to its response to ethylene. Several ethylene response mutants have been isolated by exploiting the triple response phenotype observed in dark-grown A. thaliana seedlings in the presence of ethylene1,4,5. The biosynthetic precursor for ethylene production in plants is 1-aminocyclopropane carboxylic acid (ACC)6 and is commonly used during the triple response assay to increase endogenous ethylene production that leads to the triple response phenotype1,4,5.
Although the ethylene response is widely studied in plants, the effect of exogenous ethylene on bacteria is vastly understudied despite the close association of bacteria with plants. One study reported that certain Pseudomonas strains can survive using ethylene as a sole source of carbon and energy7. However, only two studies have demonstrated that bacteria respond to ethylene. The first study showed that strains of Pseudomonas aeruginosa, P. fluorescens, P. putida, and P. syringae were chemotactic toward ethylene using an agarose plug assay in which molten agarose was mixed with a chemotaxis buffer equilibrated with pure ethylene gas8. However, to our knowledge, there have been no further reports using pure ethylene gas to characterize bacterial ethylene response, likely due to the difficultly of handling gases in the laboratory without specialized equipment. The second report of bacterial ethylene response demonstrated that ethylene increased bacterial cellulose production and influenced gene expression in the fruit-associated bacterium, Komagataeibacter (formerly Gluconacetobacter) xylinus9. In this case, the ethylene-releasing compound, 2-chloroethylphosphonic acid (CEPA) was used to produce ethylene in situ within the bacterial growth medium, bypassing the need for pure ethylene gas or specialized equipment.
CEPA produces ethylene at a 1:1 molar ratio above pH 3.510,11 through a base-catalyzed, first-order reaction12-14. The degradation of CEPA is positively correlated with pH and temperature13,14 and results in the production of ethylene, chloride and phosphate. CEPA provides researchers interested in studying bacterial responses to ethylene with a convenient alternative to gaseous ethylene.
The overall goal of the following protocols is to provide a simple and efficient method to study bacterial ethylene response and includes validation of physiologically relevant levels of ethylene production from CEPA decomposition in bacterial growth medium, analysis of culture pH to ensure CEPA decomposition is not impaired during bacterial growth, and assessment of the effect of ethylene on bacterial morphology and phenotype. We demonstrate these protocols using K. xylinus, however, these protocols can be adapted to study ethylene response in other bacteria by using the appropriate growth medium and phenotype analyses.

<table border="0" cellpadding="0" cellspacing="0" width="1018">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">1-aminocyclopropane carboxylic acid (ACC)</td>
			<td style="width:136px;">Sigma</td>
			<td style="width:119px;">A3903</td>
			<td style="width:432px;">Biosynthetic precursor of ethylene in plants</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">4-sector Petri dish</td>
			<td style="width:136px;">Phoenix Biomedical</td>
			<td style="width:119px;">CA73370-022</td>
			<td>For testing triple response</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Agar</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">AGR001.1</td>
			<td>To solidify medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Canon Rebel T1i DLSR camera</td>
			<td style="width:136px;">Canon</td>
			<td style="width:119px;">3818B004</td>
			<td>For pictures of pellicles</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Cellulase from Trichoderma reesei ATCC 26921&#160;</td>
			<td style="width:136px;">Sigma</td>
			<td style="width:119px;">C2730</td>
			<td>Aqueous solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Citric acid</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">CIT002.500</td>
			<td>For SH medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Commercial bleach</td>
			<td style="width:136px;">Life Brand</td>
			<td style="width:119px;">57800861874</td>
			<td>Bleach for seed sterilization</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Concentrated HCl</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">HCL666.500</td>
			<td>Hydrochloric acid for pH adjustment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Digital USB microscope</td>
			<td style="width:136px;">Plugable</td>
			<td style="width:119px;">N/A</td>
			<td>For pictures of colonies</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Ethephon (&#8805; 96%; 2-chloroethylphosphonic acid)</td>
			<td style="width:136px;">Sigma</td>
			<td style="width:119px;">C0143</td>
			<td>Ethylene-releasing compound</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Glucose</td>
			<td style="width:136px;">BioBasic</td>
			<td style="width:119px;">GB0219</td>
			<td>For SH medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Komagataeibacter xylinus ATCC 53582</td>
			<td style="width:136px;">ATCC</td>
			<td style="width:119px;">53582</td>
			<td>Bacterial cellulose-producing alphaproteobacterium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Microcentrifuge tube</td>
			<td style="width:136px;">LifeGene</td>
			<td style="width:119px;">LMCT1.7B</td>
			<td>1.7 mL microcentrifuge tube</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Murashige and Skoog (MS) basal medium&#160;</td>
			<td style="width:136px;">Sigma</td>
			<td style="width:119px;">M5519</td>
			<td>Arabidopsis thaliana growth medium</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:331px;">Na2HPO4&#183;7H2O&#160;</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">SPD579.500</td>
			<td>Sodium phosphate, dibasic heptahydrate for SH medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">NaCl</td>
			<td style="width:136px;">BioBasic</td>
			<td style="width:119px;">SOD001.1</td>
			<td>Sodium chloride for saline and control solution</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:331px;">NaH2PO4&#183;H2O&#160;</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">SPM306.500</td>
			<td>Sodium phosphate, monobasic monohydrate for control solution</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">NaOH</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">SHY700.500</td>
			<td>Sodium hydroxide for pH adjustment</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Paraffin film</td>
			<td style="width:136px;">Parafilm</td>
			<td style="width:119px;">PM996</td>
			<td>For sealing plates and flasks</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Peptone (bacteriological)</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">PEP403.1</td>
			<td>For SH medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Petroff-Hausser counting chamber</td>
			<td style="width:136px;">Hausser scientific</td>
			<td style="width:119px;">3900</td>
			<td>Bacterial cell counting chamber</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Polyethersulfone sterilization filter 0.2 &#181;m</td>
			<td style="width:136px;">VWR</td>
			<td style="width:119px;">28145-501</td>
			<td>For sterilizing cellulase</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Sucrose</td>
			<td style="width:136px;">BioShop</td>
			<td style="width:119px;">SUC600.1</td>
			<td>Sucrose for MS medium</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:331px;">Yeast extract</td>
			<td style="width:136px;">BioBasic</td>
			<td style="width:119px;">G0961</td>
			<td>For SH medium</td>
		</tr>
	</tbody>
</table>

utilizing the ethylene-releasing compound, 2-chloroethylphosphonic acid, as a tool to study ethylene response in bacteria

Ethylene (C2H4) is a gaseous phytohormone that is involved in numerous aspects of plant development, playing a dominant role in senescence and fruit ripening. Exogenous ethylene applied during early plant development triggers the triple response phenotype; a shorter and thicker hypocotyl with an exaggerated apical hook. Despite the intimate relationship between plants and bacteria, the effect of exogenous ethylene on bacteria has been greatly overlooked. This is partly due to the difficulty of controlling gaseous ethylene within the laboratory without specialized equipment. 2-Chloroethylphosphonic acid (CEPA) is a compound that decomposes into ethylene, chlorine, and phosphate in a 1:1:1:1 molar ratio when dissolved in an aqueous medium of pH 3.5 or greater. Here we describe the use of CEPA to produce in situ ethylene for the investigation of ethylene response in bacteria using the fruit-associated, cellulose-producing bacterium Komagataeibacter xylinus as a model organism. The protocols described herein include both the verification of ethylene production from CEPA via the Arabidopsis thaliana triple response assay and the effects of exogenous ethylene on K. xylinus cellulose production, pellicle properties and colonial morphology. These protocols can be adapted to examine the effect of ethylene on other microbes using appropriate growth media and phenotype analyses. The use of CEPA provides researchers with a simple and efficient alternative to pure ethylene gas for the routine determination of bacterial ethylene response.

A schematic plate setup for verification of ethylene liberation from CEPA in SH medium (pH 7) by the triple response assay is shown in Figure 1A-C. A flow-chart illustrating the pellicle protocol is shown in Figure 2. Dark-grown A. thaliana seedlings exhibit the triple response phenotype (shorter and thicker hypocotyl with an exaggerated apical hook) in the presence of ACC and in the presence of ethylene produced through the dec...

Watch this Scientific Journal Video about Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria at JoVE.com

Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria

The protocols outlined herein facilitate the convenient investigation of bacterial ethylene responses by utilizing 2-chloroethylphosphonic acid (CEPA). Ethylene is produced in situ through the decomposition of CEPA in an aqueous bacterial growth medium, circumventing the requirement for pure ethylene gas.

Ethylene (C2H4) is a gaseous phytohormone that is involved in numerous aspects of plant development, playing a dominant role in senescence and fruit ...

Research

JoVE Journal

Biochemistry

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions

Protein-protein interactions play an essential role in the function of a living organism. Once an interaction has been identified and validated it is ...

Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram

Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram

The serotonin transporter is a sodium and chloride-coupled transporter that &#34;pumps&#34; extracellular serotonin into cells. S-citalopram is a drug ...

Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction

The dissociation constant describes the interaction between two partners in the binding equilibrium and is a measure of their affinity. It is a crucial ...

A Colorimetric Method for Measuring Iron Content in Plants

Iron, one of the most important micronutrients in living organisms, is involved in basic processes, such as respiration and photosynthesis. Iron content ...

Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

Small-Scale Colorimetric Assays of Intracellular Lactate and Pyruvate in the Nematode Caenorhabditis elegans

Lactate and pyruvate are key intermediates of intracellular energy metabolic pathways. Monitoring the lactate/pyruvate ratio in cells helps to determine ...

Dissipative Microgravimetry to Study the Binding Dynamics of the Phospholipid Binding Protein Annexin A2 to Solid-supported Lipid Bilayers Using a Quartz Resonator

The dissipative quartz crystal microbalance technique is a simple and label-free approach to measure simultaneously the mass uptake and viscoelastic ...

Isolation of Lipoprotein Particles from Chicken Egg Yolk for the Study of Bacterial Pathogen Fatty Acid Incorporation into Membrane Phospholipids

Staphylococcus aureus and other Gram-positive pathogens incorporate fatty acids from the environment into membrane phospholipids. During infection, the ...

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

The purification of monoclonal antibodies (mAbs) is commonly achieved by Protein A affinity chromatography, which can account for up to 25% of the overall ...

In Vivo Hydroxyl Radical Protein Footprinting for the Study of Protein Interactions in Caenorhabditis elegans

In Vivo Hydroxyl Radical Protein Footprinting for the Study of Protein Interactions in Caenorhabditis elegans

Fast oxidation of proteins (FPOP) is a hydroxyl radical protein footprinting (HRPF) method used to study protein structure, protein-ligand interactions, ...

Utilizing Time-Resolved Protein-Induced Fluorescence Enhancement to Identify Stable Local Conformations One &#945;-Synuclein Monomer at a Time

Utilizing Time-Resolved Protein-Induced Fluorescence Enhancement to Identify Stable Local Conformations One ÃƒÅ½Ã‚Â±-Synuclein Monomer at a Time

Using spectroscopic rulers to track multiple conformations of single biomolecules and their dynamics have revolutionized the understanding of structural ...