Purification and Characterization of Rho 1D4 Antibody

Anti-β-actin (catalog no. 4970) was purchased from Cell Signaling Technology (MA, USA). Mouse monoclonal anti-Rho 1D4 antibody was purified from hybridoma cells^{74 (link)–76 (link)}. Alexa Fluor 647-conjugated goat anti-mouse IgG antibody (catalog no. A21236) was from Invitrogen (CA, USA). DAPI fluorescent dye (catalog no. 62248) was purchased from Thermo Fisher (MA, USA). All detergents were purchased from Anatrace (OH, USA). Hydroxylamine hydrochloride (catalog no. 159417) was purchased from Sigma-Aldrich (MI, USA). 9-cis-retinal and 11-cis-retinal were produced by photoisomerization of all-trans-retinal using 420 nm light and purified by normal-phase HPLC using a Phenomenex Luna silica column (catalog no. 00G-4091-P0-AX) and an isocratic gradient of 10% ethyl acetate in hexanes^{77 (link),78 (link)}.

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Wu A., Salom D., Hong J.D., Tworak A., Watanabe K., Pardon E., Steyaert J., Kandori H., Katayama K., Kiser P.D, & Palczewski K. (2023). Structural basis for the allosteric modulation of rhodopsin by nanobody binding to its extracellular domain. Nature Communications, 14, 5209.

Publication 2023

Anti-β-actin Alexa Fluor 647-conjugated goat anti-mouse IgG antibody DAPI fluorescent dye Hydroxylamine hydrochloride Luna silica column

11 cis retinal 9 cis retinal Actin Alexa fluor 647 Anti igg Antibody Dapi Detergents Ethyl acetate Fluorescent dye Goat Hplc Hybridoma Hydroxylamine hydrochloride Light Monoclonal antibody Mouse Retinal Silica

Corresponding Organization :

Other organizations : University of California, Irvine, Nagoya Institute of Technology, Vrije Universiteit Brussel

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Variable analysis

independent variables

9-cis-retinal and 11-cis-retinal produced by photoisomerization of all-trans-retinal using 420 nm light and purified by normal-phase HPLC

dependent variables

Not explicitly mentioned

control variables

Anti-β-actin (catalog no. 4970) from Cell Signaling Technology
Mouse monoclonal anti-Rho 1D4 antibody purified from hybridoma cells
Alexa Fluor 647-conjugated goat anti-mouse IgG antibody (catalog no. A21236) from Invitrogen
DAPI fluorescent dye (catalog no. 62248) from Thermo Fisher
All detergents purchased from Anatrace
Hydroxylamine hydrochloride (catalog no. 159417) from Sigma-Aldrich

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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