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Tamra c5 maleimide

Manufactured by Biotium
Sourced in Germany, United States

TAMRA-C5-maleimide is a fluorescent labeling reagent that can be used to covalently attach a TAMRA (Tetramethylrhodamine) fluorescent dye to proteins or other biomolecules containing a free sulfhydryl group. The maleimide functional group reacts selectively with cysteine residues, allowing for site-specific labeling of proteins.

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2 protocols using tamra c5 maleimide

1

Fab Expression and Purification

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E. coli competent DH5α cells for gene cloning and plasmid amplification
were purchased from Shenzhen Kangti Life Science (Shenzhen, China),
and E. coli SHuffle T7 express lysY from NEB31 (link),32 (link) was used for Fab expression of antidigoxin antibody. Plasmid extraction
and gel recovery kits were purchased from Vazyme Biotechnology Co.,
Ltd. (Nanjing, China). The restriction enzyme used for constructing
the recombinant plasmid was purchased from New England Biolabs (NEB;
Ipswich, MA, USA). The Ni-NTA Sefinose Resin for protein purification
was purchased from Changzhou Smart-Lifesciences Biotechnology Co.,
Ltd. (Changzhou, China), and Dig-BSA was purchased from Wuhan Yunclon
Technology Co., Ltd. (Wuhan, China). ATTO520-C2-maleimide was obtained
from ATTO-TEC (Siegen, Germany) and TAMRA-C5-maleimide was obtained
from Biotium (Hayward, CA, USA). Other reagents, if not specified,
were purchased from Sigma-Aldrich, Shanghai Sango Biotechnology, and
Beijing Solarbio Biotechnology Co. Ltd. (Beijing, China). Primers
used in this study were synthesized by Shanghai Sangong Biotechnology
Co., Ltd. (Shanghai China).
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2

Methodological Workflow for Protein Modification and Purification

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The KOD-Plus-Neo DNA polymerase and T4 ligase were obtained from Toyobo (Osaka, Japan). The restriction enzymes and the E. coli SHuffle T7 Express lysY cells were obtained from New England Biolabs Japan (Tokyo, Japan). The oligonucleotides were obtained from Operon-Eurofins (Tokyo, Japan). The PureYield plasmid miniprep kit was obtained from Promega (Tokyo, Japan). The ultrafiltration devices were obtained from Millipore (centrifugal filter tube Ultra-4, MWCO 3 k; Tokyo, Japan). The immobilized Tris(2-carboxyethyl)-phosphine (TCEP) disulfide-reducing gel was obtained from Pierce Biotechnology (Thermo Fisher Scientific, Rockford, IL, USA). ATTO520-C2-maleimide was obtained from the ATTO-TEC (Siegen, Germany). TAMRA-C5-maleimide was obtained from Biotium (Hayward, CA, USA). The Talon resin was obtained from Clontech (Takara-Bio, Shiga, Japan). The His SpinTrap column was obtained from GE Healthcare (Piscataway, NJ, USA). Anti DYKDDDDK-tag antibody beads and the DYKDDDDK peptide were obtained from Wako Pure Chemicals (Osaka, Japan). The recombinant HA protein from A/California/04/2009 H1N1 was obtained from Sino Biological (Beijing, China). Unless otherwise indicated, all other chemicals and reagents used were from Wako Pure Chemicals or Sigma (Tokyo, Japan).
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