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Iodoacetamide

Iodoacetamide is a chemical compound commonly used in biochemical research.
It is a potent alkylating agent that reacts with the sulfhydryl groups of proteins, leading to their irreversible modification.
Iodoacetamide is frequently employed in the study of protein structure and function, as well as in the identification and analysis of cysteine residues within proteins.
Researchers utilize Iodoacetamide to investigate a variety of biological processes, including signal transduction, enzyme activity, and protein-protein interactions.
This versatile reagent is also used in the development of proteomic techniques, such as mass spectrometry-based analyses, to enhance the detection and characterization of proteins.
Iodoacetamide's unique properties make it an invaluable tool for advancing our understanding of the complex mechanisms underlying cellular and molecular biology.

Most cited protocols related to «Iodoacetamide»

Quantitative Proteomics of HeLa-E. coli Mixture

Cited 284 times

An Escherichia coli K12 strain was grown in standard LB medium, harvested, washed in PBS, and lysed in BugBuster (Novagen Merck Chemicals, Schwalbach, Germany) according to the manufacturer's protocol. HeLa S3 cells were grown in standard RPMI 1640 medium supplemented with glutamine, antibiotics, and 10% FBS. After being washed with PBS, cells were lysed in cold modified RIPA buffer (50 mm Tris-HCl, pH 7.5, 1 mm EDTA, 150 mm NaCl, 1% N-octylglycoside, 0.1% sodium deoxycholate, complete protease inhibitor mixture (Roche)) and incubated for 15 min on ice. Lysates were cleared by centrifugation, and after precipitation with chloroform/methanol, proteins were resuspended in 6 m urea, 2 m thiourea, 10 mm HEPES, pH 8.0. Prior to in-solution digestion, 60-μg protein samples from HeLa S3 lysates were spiked with either 10 μg or 30 μg of E. coli K12 lysates based on protein amount (Bradford assay). Both batches were reduced with dithiothreitol and alkylated with iodoacetamide. Proteins were digested with LysC (Wako Chemicals, GmbH, Neuss, Germany) for 4 h and then trypsin digested overnight (Promega, GmbH, Mannheim, Germany). Digestion was stopped by the addition of 2% trifluroacetic acid. Peptides were separated by isoelectric focusing into 24 fractions on a 3100 OFFGEL Fractionator (Agilent, GmbH, Böblingen, Germany) as described in Ref. 45 (link). Each fraction was purified with C₁₈ StageTips (46 (link)) and analyzed via liquid chromatography combined with electrospray tandem mass spectrometry on an LTQ Orbitrap (Thermo Fisher) with lock mass calibration (47 (link)). All raw files were searched against the human and E. coli complete proteome sequences obtained from UniProt (version from January 2013) and a set of commonly observed contaminants. MS/MS spectra were filtered to contain at most eight peaks per 100 mass unit intervals. The initial MS mass tolerance was 20 ppm, and MS/MS fragment ions could deviate by up to 0.5 Da (48 (link)). For quantification, intensities can be determined alternatively as the full peak volume or as the intensity maximum over the retention time profile, and the latter method was used here as the default. Intensities of different isotopic peaks in an isotope pattern are always summed up for further analysis. MaxQuant offers a choice of the degree of uniqueness required in order for peptides to be included for quantification: “all peptides,” “only unique peptides,” and “unique plus razor peptides” (42 (link)). Here we chose the latter, because it is a good compromise between the two competing interests of using only peptides that undoubtedly belong to a protein and using as many peptide signals as possible. The distribution of peptide ions over fractions and samples is shown in supplemental Fig. S1.

Cox J., Hein M.Y., Luber C.A., Paron I., Nagaraj N, & Mann M. (2014). Accurate Proteome-wide Label-free Quantification by Delayed Normalization and Maximal Peptide Ratio Extraction, Termed MaxLFQ. Molecular & Cellular Proteomics : MCP, 13(9), 2513-2526.

Publication 2014

Acids Antibiotics, Antitubercular Biological Assay Buffers Cells Centrifugation Chloroform Cold Temperature Deoxycholic Acid, Monosodium Salt Digestion Dithiothreitol Edetic Acid Escherichia coli Escherichia coli K12 Glutamine HeLa Cells HEPES Homo sapiens Immune Tolerance Iodoacetamide Ions Isotopes Liquid Chromatography Methanol Peptides Promega Protease Inhibitors Proteins Proteome Radioimmunoprecipitation Assay Retention (Psychology) Sodium Chloride Staphylococcal Protein A Tandem Mass Spectrometry Thiourea Tromethamine Trypsin Urea

Quantitative Proteomics of Yeast Metabolism

Cited 76 times

We showcased our method using a TMT10-plex of yeast (S. cerevisiae wild-type strain BY4716) grown in synthetic complete media supplemented with 2% glucose (n=5) or 2% pyruvate (n=5) as the carbon source. We harvested the cells at OD_600nm=0.8. Cells were lysed by bead-beating in 8 M urea 200mM EPPS (4-(2-Hydroxyethyl)-1-piperazinepropanesulfonic acid), pH 8.5 and with protease and phosphatase inhibitors. Protein concentration was determined with the BCA assay. The BCA assay was performed according to manufacturer’s instructions with samples that were diluted at least 1:20, to ensure that the 8M urea has been diluted far below its compatibility limit. Samples were reduced with 5mM TCEP, alkylated with 10 mM iodoacetamide that was quenched with 10 mM DTT. A total of 100 μg of protein were chloroform-methanol precipitated. Protein was reconstituted in 200 mM EPPS pH 8.5 and digested by Lys-C overnight and trypsin for 6 h, both at a 1:100 protease-to-peptide ratio. Directly to the digest, we added a final volume of 30% acetonitrile and labelled 100 μg of peptide with 200 μg of TMT. To check mixing ratios, 2 μg of each sample were pooled, desalted, and analyzed by mass spectrometry. Using normalization factors calculated from this “label check,” samples were mixed 1:1 across all channels and desalted using a 100 mg Sep-Pak solid phase extraction column. The Pierce High-Select Fe-NTA Phosphopeptide Enrichment Kit was used to enrich phosphopeptides from the pooled TMT-labeled mixture. The unbound fraction and washes from this enrichment were combined and fractionated with basic pH reversed-phase (BPRP) HPLC, collected in a 96-well plate and combined down to 12 fractions prior to desalting and subsequent LC-MS/MS processing (14 (link), 15 ).

Navarrete-Perea J., Yu Q., Gygi S.P, & Paulo J.A. (2018). SL-TMT: A Streamlined Protocol for Quantitative (Phospho)proteome Profiling using TMT-SPS-MS3. Journal of proteome research, 17(6), 2226-2236.

Publication 2018

acetonitrile Acids Biological Assay Carbon Cells Chloroform ferric nitrilotriacetate Glucose High-Performance Liquid Chromatographies inhibitors Iodoacetamide Mass Spectrometry Methanol Peptide Hydrolases Peptides Phosphopeptides Phosphoric Monoester Hydrolases Proteins Pyruvate Saccharomyces cerevisiae Solid Phase Extraction Staphylococcal Protein A Strains Tandem Mass Spectrometry tris(2-carboxyethyl)phosphine Trypsin Urea Yeast, Dried

Histone H4 cysteine mapping

Cited 65 times

We genetically engineered S. cerevisiae to contain a cysteine at position 47 in histone H4. Cells grown to mid-log phase were harvested, permeabilized and labeled with N(1,10 phenanthroline- 5-yl) iodoacetamide. The label covalently bound to the cysteine and allowed for copper chelation. Copper chloride, mercaptoproprionic acid and hydrogen peroxide were added sequentially creating hydroxyl radicals that cleaved the nucleosomal DNA at sites flanking the center. After the mapping reaction, the genomic DNA was purified from the cells and ran on an agarose gel. The shortest molecular weight DNA fragment (~150-200bp) was purified and prepared for highthroughput parallel sequencing.

Brogaard K., Xi L., Wang J.P, & Widom J. (2012). A base pair resolution map of nucleosome positions in yeast. Nature, 486(7404), 496-501.

Publication 2012

Acids Cells Chlorides Copper Cysteine Genome Histone H4 Hydroxyl Radical Iodoacetamide Nucleosomes Peroxide, Hydrogen Phenanthrolines Sepharose

Histone H4 cysteine mapping

Cited 61 times

Brogaard K., Xi L., Wang J.P, & Widom J. (2012). A base pair resolution map of nucleosome positions in yeast. Nature, 486(7404), 496-501.

Publication 2012

Acids Cells Chlorides Copper Cysteine Genome Histone H4 Hydroxyl Radical Iodoacetamide Nucleosomes Peroxide, Hydrogen Phenanthrolines Sepharose

Protein Mixture Alkylation and Tryptic Digestion

Cited 58 times

A standard mixture was prepared by suspending in
200 mM ABC the proteins α-casein (bovine), β-casein (bovine),
enolase (yeast), apo-transferrin (human), carbonic
anhydrase (bovine), and ribonuclease B (bovine) to concentrations
of 6, 2, 4, 2.3, 2.5, and 2 μg/μL. Eight 400 μg
aliquots of this mixture were alkylated and digested as described
later (Table 1B).
Aliquots suspended
in the pH 8 reducing buffers specified in Table 1B were incubated at 50 °C with shaking for 60 min, after which
they were alkylated with 30 mM iodoacetamide (IAN) or 25 mM 4-VP,
shaking for 30 min at 37 °C.
All alkylated samples were
quenched by the addition of 200 mM DTT
to a final concentration of 22 mM and then diluted 1:1 with either
25 mM ABC or 0.1% DCA in 25 mM ABC (Table 1B). Modified, sequencing-grade trypsin (Promega) was added to each
sample (1:30 w/w). Digestion proceeded for 12 h on a 37 °C shaker.
Aliquots (10 μg) were removed for SDS-PAGE analysis.

Erde J., Loo R.R, & Loo J.A. (2014). Enhanced FASP (eFASP) to Increase Proteome Coverage and Sample Recovery for Quantitative Proteomic Experiments. Journal of Proteome Research, 13(4), 1885-1895.

Publication 2014

ABCA1 protein, human Bos taurus Buffers Caseins Digestion Enolase Homo sapiens Iodoacetamide Promega Proteins ribonuclease B SDS-PAGE Transferrin Trypsin Yeast, Dried

Most recents protocols related to «Iodoacetamide»

Proteomic Analysis of Lysosomal Enzymes

25 µg of purified AGA, GUSB CTSD, and GAA were dissolved in 50 mM ammonium bicarbonate (AmBic) buffer (pH 7.4) and further reduced with 10 mM dithiothreitol (DTT) at 60°C for 45 min on shaker, followed by alkylation with 20 mM iodoacetamide (IAA) at 25°C for 30 min in darkness. AGA, GUSB, CTSD were subjected to proteolytic digestion with chymotrypsin (1:40 enzyme-substrate ratio), while GAA was digested in gel with trypsin (1:25 enzyme-substrate ratio) after SDS-PAGE separation. The reaction was quenched with 1 µL trifluoroacetic acid (TFA) and the digested sample was desalted by custom-made modified StageTip colums with three layers of C18 and two layers of C8 membrane (3 M Empore disks, Sigma-Aldrich). Samples were eluted with two steps of 50 µL 50% methanol in 0.1% formic acid. Final sample was aliqoted in two equal parts. The first aliquot was placed into a glass insert (Agilent), dried completely in SpeedVac (Eppendorf) and further re-dissolved in 50 µL 0.1% formic acid (FA) and submitted for nLC-MS analysis. The second aliqout was placed inside an Eppendorf tube, dried completely using SpeedVac, and then re-dissolved in 50 µL of 50 mM AmBic buffer (pH 7.4) and incubated with PNGase F (1U per sample) for 12 h with shaking at 37°C. Samples treated with PNGase F were desalted and dried using the same methods mentioned above for the first aliqout and submitted for nLC-MS/MS analysis.

Chen Y.H., Tian W., Yasuda M., Ye Z., Song M., Mandel U., Kristensen C., Povolo L., Marques A.R., Čaval T., Heck A.J., Sampaio J.L., Johannes L., Tsukimura T., Desnick R., Vakhrushev S.Y., Yang Z, & Clausen H. (2023). A universal GlycoDesign for lysosomal replacement enzymes to improve circulation time and biodistribution. Frontiers in Bioengineering and Biotechnology, 11, 1128371.

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Publication 2023

Alkylation ammonium bicarbonate Buffers Chymotrypsin CTSD protein, human Darkness Digestion Dithiothreitol Empore Enzymes formic acid Glycopeptidase F Iodoacetamide Methanol Peptide Hydrolases SDS-PAGE Tandem Mass Spectrometry Tissue, Membrane Trifluoroacetic Acid Trypsin

Protein Extraction and Trypsin Digestion of L. paraplantarum RX-8

Cell samples of L. paraplantarum RX-8 in co-culture and mono-culture were collected at 24 h according to Section 2.5.1. The protein was extracted by using a lysis buffer (8 M urea, 50 mM Tris8.0, 1% NP40, 1% sodium deoxycholate, 5 mM dithiothreitol (DTT), 2 mM EDTA, 30 mM nicotinamide, and 3 μm trichostatin A), and, after sonication on ice, the total protein concentration of the supernatant, which was obtained by centrifugation (20,000 rpm, 10 min, 4°C), was determined by using a BCA Protein Assay kit. The protein sample was reduced by DTT (5 mM, 45 min, 30°C), later alkylated with 30 mM iodoacetamide (30 mM, 1 h, RT) in darkness, and then precipitated with ice-cold acetone. After being washed thrice with acetone, the precipitate was suspended in 0.1 M triethylammonium bicarbonate (TEAB) and digested with trypsin (1/25 protein mass, Promega) for 12 h at 37°C. Finally, the reaction was ended with 1% trifluoroacetic acid (TFA), and the resulting peptide was desalted with Strata X C18 SPE column (Phenomenex, Torrance, CA, USA) and vacuum-dried in Scanvac maxi-beta (Labogene, Alleroed, Denmark).

Nie R., Zhu Z., Qi Y., Wang Z., Sun H, & Liu G. (2023). Bacteriocin production enhancing mechanism of Lactiplantibacillus paraplantarum RX-8 response to Wickerhamomyces anomalus Y-5 by transcriptomic and proteomic analyses. Frontiers in Microbiology, 14, 1111516.

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Publication 2023

Acetone Biological Assay Buffers Centrifugation Coculture Techniques Cold Temperature Darkness Deoxycholic Acid, Monosodium Salt Dithiothreitol Edetic Acid Iodoacetamide L Cells Niacinamide Peptides Promega Proteins trichostatin A triethylammonium bicarbonate Trifluoroacetic Acid Trypsin Urea Vacuum

Paramagnetic Bead-based Protein Cleanup

For protein cleanup, the paramagnetic bead–based SP3 (solid-phase–enhanced sample preparation) workflow was used (Hughes et al, 2019 (link)). For each AP experiment, sample protein concentrations were determined using the Pierce BCA protein assay (Thermo Fisher Scientific) following the manufacturer’s instructions, and 50 μg of proteins was adjusted in 20 μl of buffer/MS-grade water. Samples were homogenized and denatured in urea (final concentration, 4 M), ammonium bicarbonate (100 mM), and calcium chloride (100 mM), then reduced in DTT (final concentration, 1 mM) for 15 min at room temperature, and alkalinized in iodoacetamide (3 mM) in the dark at room temperature for 15 min. The tryptic digestion protocol was performed using the KingFisher DuoPrime purification system (Thermo Fisher Scientific) in a series of steps. First, magnetic hydrophobic and hydrophilic beads were washed several times in MS-grade water and added to the deepwell plate in the KingFisher along with the samples and the same volume as the sample of 100% ethanol. Next, the solutions were mixed at low speed for 10 min, after which the beads coupled to the proteins were collected with the magnetic arm of the KingFisher and transferred to be washed in three different deepwells each containing 80% of ethanol. The washed beads–proteins were then released into the trypsin (V5111; Promega)-containing deepwells at a 50:1 (w/w) protein-to-protease ratio and mixed at low speed for 8 h of digestions into peptide fragments at 37°C in the KingFisher. Peptide samples were transferred into low protein binding tubes; 1% of TFA was added to acidify the samples ready to be desalted, cleaned, and concentrated on C18 tips (87784; Thermo Fisher Scientific) (Rappsilber et al, 2007 (link)) according to the manufacturer’s instructions. Purified peptides were dried and resuspended in low protein binding tubes before MS analysis in 30 μl of 0.15% TFA and 1% acetic acid in MS-grade water.

Ternet C., Junk P., Sevrin T., Catozzi S., Wåhlén E., Heldin J., Oliviero G., Wynne K, & Kiel C. (2023). Analysis of context-specific KRAS–effector (sub)complexes in Caco-2 cells. Life Science Alliance, 6(5), e202201670.

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Publication 2023

Acetic Acid ammonium bicarbonate Biological Assay Buffers Calcium chloride Diet, Protein-Restricted Digestion Ethanol Iodoacetamide Peptide Fragments Peptide Hydrolases Peptides Promega Proteins Trypsin Urea

Synchronizing C. elegans for Phosphoproteomics

To obtain synchronized young adults, five to six L4 larvae animals were picked onto standard 60-mm culture plate spread with OP50 bacteria. Animals were grown at 20°C for 5 days until starved. 90 plates of L1 larvae for each genotype or condition were washed into 1.5-l liquid culture supplemented with HB101 bacteria. Worms were then grown with aeration at 200 rpm at 20°C for 3 days to reach adulthood. Six hours prior to harvest, 1 mM auxin or 0.25% ethanol (solvent control) was added to the liquid culture. Harvested worms were frozen in liquid nitrogen and stored at −80°C. The frozen worms were then processed using a prechilled Retsch mixer mill to break the cuticle, thawed on ice in cold lysis buffer (25 mM HEPES pH 7.4, 100 mM NaCl, 1 mM MgCl₂, 1 mM EGTA, 0.1% Triton X-100, 1 mM DTT, cOmplete protease inhibitors [Sigma #4693159001] and phosSTOP [Sigma #4906837001]). Lysates were processed using a Dounce homogenizer and sonicated using a Branson Digital Sonifier on ice and then centrifuged at 20,000 RCF for 25 min at 4°C. The supernatant was incubated with anti-ALFA selector (Nanotag Biotechnologies, #N1511) for 3 hr at 4°C. Beads were then washed six times with lysis buffer and three times with milli-Q water. Proteins on beads were then processed for phosphorylation site identification using mass spectrometry (UC Davis).
Briefly, beads were spun in a 10K MWCO filter (VWR, Radnor, PA) at room temperature for 10 min at 10,000 × g and then washed with 50 mM ammonium bicarbonate. Beads were then subjected to reduction at 56°C for 45 min in 5.5 mM DTT followed by alkylation for 1 hr in the dark with iodoacetamide added to a final concentration of 10 mM. The beads were again washed with 50 mM ammonium bicarbonate followed by addition of sequencing grade trypsin to a final enzyme:substrate mass ratio of 1:50 and digested overnight at 37°C. Resultant peptides were then collected in a fresh clean centrifuge tube during a final spin of 16,000 × g for 20 min. Peptides were dried down in a speed-vac and stored at −80°C. Prior to analysis, samples were reconstituted in 2% acetonitrile with 0.1% TFA. Samples were then loaded and analyzed as described above.

Zhang L., Stauffer W.T., Wang J.S., Wu F., Yu Z., Liu C., Kim H.J, & Dernburg A.F. (2023). Recruitment of Polo-like kinase couples synapsis to meiotic progression via inactivation of CHK-2. eLife, 12, e84492.

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Publication 2023

acetonitrile Alkylation ammonium bicarbonate Animals Auxins Bacteria Buffers Cold Temperature Egtazic Acid Enzymes Ethanol Freezing Genotype Helminths HEPES Iodoacetamide Larva Magnesium Chloride Mass Spectrometry Nitrogen Peptides Protease Inhibitors Proteins Sodium Chloride Solvents Triton X-100 Trypsin Young Adult

SILAC-Based Protein Quantification and Digestion

The protein concentration of the SILAC cell lysates was determined using the bicin chonicic acid assay (Pierce). Digestion of the proteins was performed using the Filter-Aided Sample Preparation (FASP) method [108 (link)], for which equal amounts (900 μg) of mock- and virus-infected cell lysates were mixed and DTT was added to a final concentration of 50 mM, followed by a 5-min incubation at 70°C. Samples were loaded on two 15-ml 30 kDa Microcon filter devices (Millipore), which were washed twice with 8 M urea 0.1 M Tris pH 8.5, while cysteines were alkylated with 50 mM iodoacetamide in the same buffer. Samples were washed 3 times with 8 M urea, 0.1 M Tris pH 8. Proteins were digested overnight at room temperature using 20 ug endoLysC (Wako Pure Chemical Industries) in the same buffer per filter device. The sample was diluted fourfold with 50 mM ammonium bicarbonate pH 8.4 containing 20 ug trypsin (Worthington Chemical Corporation), and digested for 4 h at room temperature. Peptides were collected by centrifugation, acidified to a final percentage of 1% TFA, and desalted using solid phase extraction. Peptides were eluted in 20/80/0.1 (v/v/v) of milliQ/acetonitrile (ACN) (Actu-All Chemicals)/trifluoric acid (TFA) (Sigma-Aldrich).

Treffers E.E., Tas A., Scholte F.E., de Ru A.H., Snijder E.J., van Veelen P.A, & van Hemert M.J. (2023). The alphavirus nonstructural protein 2 NTPase induces a host translational shut-off through phosphorylation of eEF2 via cAMP-PKA-eEF2K signaling. PLOS Pathogens, 19(2), e1011179.

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Publication 2023

acetonitrile Acids ammonium bicarbonate Biological Assay Buffers Cells Centrifugation Cysteine Iodoacetamide Medical Devices Peptides Protein Digestion Proteins Solid Phase Extraction Tromethamine Trypsin Urea Virus

Top products related to «Iodoacetamide»

Trypsin by Promega

Sourced in United States, Germany, United Kingdom, China, Japan, France, Switzerland, Sweden, Italy, Netherlands, Spain, Canada, Brazil, Australia, Macao

Trypsin is a serine protease enzyme that is commonly used in cell culture and molecular biology applications. It functions by cleaving peptide bonds at the carboxyl side of arginine and lysine residues, which facilitates the dissociation of adherent cells from cell culture surfaces and the digestion of proteins.

Iodoacetamide by Merck Group

Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, Italy, France, Macao, China, Canada, Australia, Switzerland, Spain

Iodoacetamide is a chemical compound commonly used in biochemistry and molecular biology laboratories. It is a reactive compound that selectively modifies cysteine residues in proteins, thereby allowing for the study of protein structure and function. Iodoacetamide is often used in sample preparation procedures for mass spectrometry and other analytical techniques.

Dithiothreitol (dtt) by Merck Group

Sourced in United States, Germany, United Kingdom, Italy, Sao Tome and Principe, France, China, Switzerland, Macao, Spain, Australia, Canada, Belgium, Sweden, Brazil, Austria, Israel, Japan, New Zealand, Poland, Bulgaria

Dithiothreitol (DTT) is a reducing agent commonly used in biochemical and molecular biology applications. It is a small, water-soluble compound that helps maintain reducing conditions and prevent oxidation of sulfhydryl groups in proteins and other biomolecules.

Sequencing grade trypsin by Promega

Sourced in United States, Germany, United Kingdom, Italy, France, Spain, Switzerland

Sequencing grade trypsin is a proteolytic enzyme used to cleave peptide bonds in protein samples, primarily for use in protein sequencing applications. It is purified to ensure high-quality, consistent performance for analytical processes.

Sequencing grade modified trypsin by Promega

Sourced in United States, United Kingdom, Germany, France, Switzerland, Japan, China, Sweden

Sequencing grade modified trypsin is a protease enzyme used for the digestion of proteins prior to mass spectrometry analysis. It is designed to provide consistent, high-quality peptide digestion for protein identification and characterization.

Ammonium bicarbonate by Merck Group

Sourced in United States, Germany, Italy, United Kingdom, France, Sao Tome and Principe, China, Spain, Macao, Australia, Switzerland, Hungary, India, Sweden

Ammonium bicarbonate is a chemical compound with the formula (NH4)HCO3. It is a white crystalline solid that is commonly used as a leavening agent in baking and as a source of carbon dioxide in certain industrial processes.

Formic acid by Merck Group

Sourced in Germany, United States, Italy, United Kingdom, France, Spain, China, Poland, India, Switzerland, Sao Tome and Principe, Belgium, Australia, Canada, Ireland, Macao, Hungary, Czechia, Netherlands, Portugal, Brazil, Singapore, Austria, Mexico, Chile, Sweden, Bulgaria, Denmark, Malaysia, Norway, New Zealand, Japan, Romania, Finland, Indonesia

Formic acid is a colorless, pungent-smelling liquid chemical compound. It is the simplest carboxylic acid, with the chemical formula HCOOH. Formic acid is widely used in various industrial and laboratory applications.

Urea by Merck Group

Sourced in United States, Germany, United Kingdom, Italy, India, France, Sao Tome and Principe, China, Australia, Spain, Poland, Belgium, Switzerland, Canada, Mexico, Macao, Pakistan, Croatia, Finland, Czechia, Chile, Sweden

Urea is a chemical compound with the formula CO(NH2)2. It is a colorless, odorless, and crystalline solid that is highly soluble in water. Urea's core function is to serve as a source of nitrogen and a key component in many biochemical processes.

Trypsin by Merck Group

Sourced in United States, Germany, United Kingdom, China, Italy, Japan, Sao Tome and Principe, Canada, Macao, Poland, India, France, Spain, Portugal, Australia, Switzerland, Ireland, Belgium, Sweden, Israel, Brazil, Czechia, Denmark, Austria

Trypsin is a serine protease enzyme that is commonly used in cell biology and biochemistry laboratories. Its primary function is to facilitate the dissociation and disaggregation of adherent cells, allowing for the passive release of cells from a surface or substrate. Trypsin is widely utilized in various cell culture applications, such as subculturing and passaging of adherent cell lines.

C18 ziptip by Merck Group

Sourced in United States, Germany, United Kingdom, China, Morocco, Canada, France, Japan, Italy

C18 ZipTips are a type of pipette tip used in sample preparation for mass spectrometry and other analytical techniques. They are coated with a C18 reversed-phase material, which can selectively retain and concentrate analytes of interest from complex biological samples.

What are the unique properties of Iodoacetamide that make it a valuable research tool?

Iodoacetamide is a versatile and powerful reagent used extensively in biochemical research. Its key properties include: 1. Potent alkylating agent: Iodoacetamide reacts irreversibly with the sulfhydryl groups of proteins, leading to the modification of cysteine residues. This makes it a valuable tool for studying protein structure and function. 2. Enabling proteomic analyses: Iodoacetamide is widely used in mass spectrometry-based proteomic techniques to enhance the detection and characterization of proteins by preventing cysteine oxidation and improving sample preparation. 3. Investigating biological processes: Researchers leverage Iodoacetamide to investigate a variety of cellular and molecular mechanisms, such as signal transduction, enzyme activity, and protein-protein interactions. 4. Versatility in applications: Due to its unique reactivity, Iodoacetamide finds applications in diverse areas of biochemistry, including the study of protein structure, post-translational modifications, and the development of new analytical methods.

What are some common challenges or considerations when using Iodoacetamide in research?

When working with Iodoacetamide, researchers should be aware of the following considerations: 1. Reaction specificity: While Iodoacetamide primarily targets cysteine residues, it can also react with other nucleophilic groups, such as histidine, lysine, and tyrosine, leading to potential off-target effects. Careful optimization of reaction conditions is necessary. 2. Irreversible modification: The alkylation of cysteine residues by Iodoacetamide is an irreversible process. This can impact downstream analyses, particularly when studying protein dynamics or reversible modifications. 3. Toxicity and safety: Iodoacetamide is a potent alkylating agent and is considered a hazardous chemical. Proper safety precautions, such as the use of personal protective equipment and proper disposal, must be observed when handling Iodoacetamide. 4. Interference with certain analyses: Iodoacetamide-modified proteins may exhibit altered physicochemical properties, which can interfere with some analytical techniques, such as electrophoresis or mass spectrometry. Researchers should be mindful of these potential impacts.

How can PubCompare.ai help researchers optimize the use of Iodoacetamide in their studies?

PubCompare.ai is an invaluable tool for researchers working with Iodoacetamide. The platform's AI-powered capabilities can assist in several ways: 1. Efficient protocol screening: PubCompare.ai allows you to quickly and efficiently screen through the vast literature on Iodoacetamide protocols, saving time and effort. 2. Identification of critical insights: The platform's AI analysis can pinpoint key differences in protocol effectiveness, enabling you to identify the most optimal approach for your specific research goals and ensure reproducibility. 3. Comparative analysis: PubCompare.ai provides insightful comparisons between different Iodoacetamide protocols, helping you make informed decisions on the best method to use based on your experimental needs and the desired outcomes. 4. Streamlined research optimization: By leveraging PubCompare.ai, you can elevate the accuracy and reproducibility of your Iodoacetamide research, taking your studies to new heights and advancing your understanding of the complex mechanisms involved.

Are there any different variations or types of Iodoacetamide available for research purposes?

Yes, there are several variations and types of Iodoacetamide that researchers can utilize, depending on their specific needs: 1. Standard Iodoacetamide: This is the most commonly used form of Iodoacetamide, which is a potent alkylating agent that reacts with sulfhydryl groups. 2. Deuterated Iodoacetamide: This variant incorporates deuterium atoms, which can be useful for mass spectrometry-based studies, providing improved resolution and sensitivity. 3. Fluorescent-labeled Iodoacetamide: Iodoacetamide can be coupled with fluorescent dyes, enabling the visualization and tracking of labeled proteins in cellular and biochemical assays. 4. Affinity-tagged Iodoacetamide: Iodoacetamide can be modified with affinity tags, such as biotin, to facilitate the capture and purification of labeled proteins for further analysis.

More about "Iodoacetamide"

Iodoacetamide (IAA) is a versatile chemical compound widely used in biochemical research.
It is a potent alkylating agent that reacts with the sulfhydryl (thiol) groups of proteins, leading to their irreversible modification.
This property makes Iodoacetamide an invaluable tool for studying protein structure, function, and interactions.
Researchers employ Iodoacetamide in a variety of applications, including the investigation of signal transduction pathways, enzyme activity, and protein-protein interactions.
It is frequently used in conjunction with other reagents, such as Trypsin, Dithiothreitol (DTT), and Ammonium bicarbonate, to enhance protein analysis and characterization techniques.
Iodoacetamide is particularly useful in the field of proteomics, where it is commonly used in mass spectrometry-based analyses.
By alkylating cysteine residues, Iodoacetamide helps improve the detection and identification of proteins, as well as the characterization of post-translational modifications (PTMs).
In addition to its applications in protein research, Iodoacetamide is also used in the development of various biochemical methods and techniques.
For example, it is employed in the preparation of Sequencing grade trypsin and Sequencing grade modified trypsin, which are essential tools for protein digestion and peptide analysis.
Furthermore, Iodoacetamide is often used in conjunction with other reagents, such as Formic acid, Urea, and C18 ZipTips, to enhance sample preparation and chromatographic separation in proteomic workflows.
The versatility and unique properties of Iodoacetamide make it an indispensable reagent for researchers seeking to advance their understanding of cellular and molecular biology.
With its ability to modify proteins and enable the investigation of various biological processes, Iodoacetamide continues to be a valuable tool in the pursuit of scientific discoveries.