Quantification of reactive oxygen species production by the red fluorescent proteins KillerRed, SuperNova and mCherry.
by Angelica
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fluorescent proteins can produce reactive oxygen species (ROS) on the absorption of photons through type I and II photosensitization mechanisms. KillerRed red fluorescent protein and Supernova are phototoxic protein engineered to produce ROS and used in a variety of biological applications. However, their relative quantum results and the level of ROS production is unclear, which has limited the interpretation of their effects when used in a biological system.
We cloned and purified KillerRed, Supernova, and mCherry – red fluorescent protein linked not usually considered photosensitizer – and measuring superoxide (O2 • -) and singlet oxygen (1O2) quantum results with irradiation at 561 nm. The formation of O2 • 2-hydroxyethidium –specific product (2-OHE +) was quantified by HPLC with fluorescence detection separation. In connection with the reference photosensitizer, Rose Bengal, O2 • – quantum results (ΦO2 • -) of Supernova was determined to be 1.5 × 10-3, KillerRed is 0.97 × 10-3 and 1.2 × 10-3 mCherry , At 916.5 excitation fluence J / cm2 and suitable absorption at 561 nm, Supernova, KillerRed and mCherry made 3.81, 2.38 and 1.65 pM O2 • – / min, respectively.
Using Singlet Oxygen Sensor probe Green (SOSG), we confirmed the results of quantum 1O2 (Φ1O2) for Supernova be 22.0 × 10-3, KillerRed 7.6 × 10-3 and 5.7 × 10-3 mCherry. This photosensitization characteristics of Supernova, KillerRed and mCherry improve our understanding of the relevant fluorescent protein and to improve their use as a tool for advancing our knowledge of redox biology.
Quantification of reactive oxygen species production by the red fluorescent proteins KillerRed, SuperNova and mCherry.
Green Fluorescent protein-and Discosoma sp. Red Fluorescent Protein-Tagged Lines for Protein Marker organelle subcellular localization in Rice.
Subcellular localization of proteins is a fundamental aspect of protein function. Determine the subcellular localization is important to understand the biological function of the protein. Here, we developed a set of rice organelle marker lines, which express the organelle fluorescent marker can be used as a standard of comparison in determining the subcellular localization of the protein of interest. We build green fluorescent protein (GFP) – and / or Discosoma sp. red fluorescent protein (DsRed) -tagged organelle markers targeted to the endoplasmic reticulum (ER), mitochondria, Golgi apparatus, peroxisome, actin cytoskeleton, plastids, tonoplast, plasma membrane and nucleus, respectively.
The utility of the marker lines of rice for subcellular protein localization studies indicated by detecting OsWRKY45 local nucleus and mitochondria-related NbHxk1 in protoplasts of GFP-OsH2B and ScCOX4 line-DsRed, respectively. Using the sheath-inoculation method, followed by the live-cell imaging, we detect the co-localization of Magnaporthe oryzae PWL2: mCherry: NLS fusion with GFP marker-OsH2B core in rice epidermal cells, confirming the translocation of M. oryzae effectors into host cells PWL2 , and further demonstrates the feasibility of using line organelle markers for studying the dynamics of proteins in the cell of rice in the rice and the interactions between pathogens.
Set line organelle markers developed in this study provides a valuable resource for the study of subcellular localization of proteins in the protein fluorescent bioimaging rice.Photoconvertible have significant power and biomedical advances. However, the molecular mechanisms for light-induced processes have remained elusive because of the challenge of tracking chromophore structural dynamics during photoconversion.
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: Rainbow Calibration Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: Rainbow Fluorescent Particlesare calibration particles that help you to determine if a flow cytometer is properly aligned and has a clean flow cell with no fluidic blockage. Using SPHERO? Alignment Particles, the coefficients of variation (CVs), peak channels, and histogram distributions can be measured to determine the alignment and functionality of the flow cytometer.
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogen and conjugated to HRP. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogen and conjugated to HRP. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP Tag from Null. This RFP Tag antibody is for WB. It is affinity-purified from mouse ascites by affinity-chromatography using the specific immunogen and conjugated to HRP. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A polyclonal antibody against RFP-Tag. Recognizes RFP-Tag from . This antibody is Unconjugated. Tested in the following application: WB;WB:1:5000
Description: Affinity purified Anti-GFP (Green Fluorescent Protein) Tag clone GF28R, mouse IgG1. Recognizes native and denatured forms of GFP and its variants such as: EGFP, YFP, EYFP, and CFP. Applications: Western Blot(1:1000-3000), Dot Blot, ELISA, Immunocytochemistry(1:500-2000), Immunoprecipitation. Optimal dilutions/concentrations should be determined by the end user.
Description: Affinity purified monoclonal Anti-GST-Tag, clone GST.B6, mouse IgG2a. Recognizes native and denatured forms of purified GST or GST fusion proteins. Applications: Western Blot (1:1000-3000), Dot Blot, ELISA, Immunocytochemistry (1:500-2000), Immunoprecipitation. Optimal dilutions/concentrations should be determined by the end user.
Description: Affinity purified anti-HA-tag clone HA.C5/C10, mouse IgG3. Recognizes the N-terminal, C-terminal or internal HA- tagged fusion proteins. Applications: Western Blot (1:1000), Dot Blot, ELISA, Immunocytochemistry (1:200), Immunoprecipitation (1:200), In Situ Hybridisation (1:500-2000). Optimal dilutions/concentrations should be determined by the end user.
Description: Affinity purified monoclonal Anti-His-Tag, clone HIS.H8/ EH158, mouse IgG2b. Recognizes His-tagged recombinant proteins or His-tagged proteins overexpressed in cells. Applications: Western blotting (1:1000), ICC (1:200), IP (1:200), ELISA, Electrophoretic mobility shift assay. Optimal dilutions/concentrations should be determined by the end user.
Description: Affinity purified Anti-Myc, clone Myc.A7/M19, Mouse IgG1. Recognizes both N-terminal or C-terminal Myc-tagged fusion proteins. Applications: Western Blot (1:1000 - 3000), Dot Blot, ELISA, Immunocytochemistry (1:500-2000), Immunoprecipitation (1:200), In Situ Hybridization. Optimal dilutions/concentrations should be determined by the end user.
Description: Mouse Anti-His Tag Monoclonal Antibody is Protein A purified and provided at 1 mg/mL. Suitable for Western blot or Immunostaining. 100 µg.
Description: Mouse Anti-FLAG Tag Monoclonal Antibody is Protein A purified and provided at 1 mg/mL. Suitable for Western blot or Immunostaining. 100 µg.
Description: Mouse Anti-GST Tag Monoclonal Antibody is Protein A purified and provided at 1 mg/mL. Suitable for Western blot or Immunostaining. 100 µg.
Description: Mouse Anti-Myc Tag Monoclonal Antibody is Protein A purified and provided at 1 mg/mL. Suitable for Western blot or Immunostaining. 100 µg.
Description: A polyclonal antibody against RFP-Tag Polyclonal. Recognizes RFP-Tag Polyclonal from N/A. This antibody is Unconjugated. Tested in the following application: WB;WB:1:2000-5000
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A monoclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from mouse antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in mouse by using as an immunogen recombinant protein
Description: A polyclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from rabbit antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen recombinant protein
Description: A polyclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from rabbit antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen recombinant protein
Description: A polyclonal antibody for detection of RFP-Tag from Null. This RFP-Tag antibody is for WB. It is affinity-purified from rabbit antiserum by affinity-chromatography using the specific immunogenand is unconjugated. The antibody is produced in rabbit by using as an immunogen recombinant protein
Description: Affinity purified Monoclonal Anti-β-Tag, clone BA3R, mouse IgG2b. Recognizes native and denatured forms of β-Actin (~42kDa). Applications: Western Blot(1:1000-10000) ,Dot Blot, ELISA, Immunocytochemistry(1:500-2000), Flow cytometry (0.1-1 µg for 10(6) cells). Optimal dilutions/concentrations should be determined by the end user.
Description: A Mouse Monoclonal antibody against T7-Tag. This antibody is tested and validated for WB, ELISA
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We apply time-resolved electronic and stimulated Raman spectroscopy to reveal the two species are hidden from an engineering ancestral GFP-like LEA proteins, involving semi-stuck protonated (GA ‘) and stuck deprotonated (GB’) chromophore trip to photoconversion pH = 7, 9 buffer.
