Peptide Synthesis and Fluorescent Labeling

Peptide 1D4 (TETSQVAPA) was synthesized at the Keck Biotechnology Resource Laboratory at Yale University. Two fluorescently labeled peptides, GLP-1-(7–37) and Ex-4, were also obtained from there with the E21K and L21K mutations, respectively for lysine conjugation with 5(6)-carboxyfluorescein (FAM). The following materials were purchased from indicated sources: peptide GLP-1-(7–37) from GL Biochem (Shanghai) Ltd; Ex-4 from Abcam; BODIPY-FL-GTPγS from Invitrogen; 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) from Avanti Polar Lipids; n-dodecyl-β-D-maltopyranoside (DDM) from Anatrace; Bio-Beads SM-2 from Bio-Rad; and Rho 1D4 purified monoclonal antibody from University of British Columbia. The antibody was coupled to UltraLink Hydrazide resin purchased from Thermo Scientific [36 (link), 37 (link)]. All other chemicals were analytical grade obtained from Sigma or American Biochemicals. Membrane scaffold protein (MSP1E3D1) was expressed and purified as described previously [33 , 35 (link)].

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Cai Y., Liu Y., Culhane K.J., DeVree B.T., Yang Y., Sunahara R.K, & Yan E.C. (2017). Purification of family B G protein-coupled receptors using nanodiscs: Application to human glucagon-like peptide-1 receptor. PLoS ONE, 12(6), e0179568.

Publication 2017

BODIPY-FL-GTPγS 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) n-dodecyl-β-D-maltopyranoside (DDM) Bio-Beads SM-2 UltraLink Hydrazide resin

1 palmitoyl 2 oleoyl sn glycero 3 phosphocholine Antibody Bodipy Carboxyfluorescein Glp 1 Hydrazide Lipids Lysine Membrane protein Monoclonal antibody Mutations Peptide Resin

Corresponding Organization : University of California, San Diego

Top 5 similar protocols

Variable analysis

independent variables

Peptide 1D4 (TETSQVAPA) synthesized at the Keck Biotechnology Resource Laboratory at Yale University
GLP-1-(7–37) peptide with E21K mutation
Ex-4 peptide with L21K mutation

dependent variables

Lysine conjugation with 5(6)-carboxyfluorescein (FAM)

control variables

BODIPY-FL-GTPγS from Invitrogen
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) from Avanti Polar Lipids
N-dodecyl-β-D-maltopyranoside (DDM) from Anatrace
Bio-Beads SM-2 from Bio-Rad
Rho 1D4 purified monoclonal antibody from University of British Columbia
Membrane scaffold protein (MSP1E3D1) expressed and purified as described previously

controls

Positive control: Not specified
Negative control: Not specified

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