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Wizard precipitant synergy block 2

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The Wizard Precipitant Synergy block #2 is a laboratory equipment designed to facilitate the process of protein crystallization. It is a specialized block that allows for the simultaneous screening of various precipitant solutions, which are crucial in the crystallization of proteins. The Wizard Precipitant Synergy block #2 provides a convenient and efficient way to explore different combinations of precipitants, helping researchers to identify the optimal conditions for protein crystal growth.

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Lab products found in correlation

Crystal screen ht, by Hampton Research (3 mentions) Clear strategy screens 1, by Hampton Research (2 mentions) Proplex screen ht 96, by Hampton Research (1 mentions)

3 protocols using wizard precipitant synergy block 2

1

Protein Complex Crystallization Conditions

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Complexes of iv1 scFv/iGL-VRC01 Fab, iv4-HL Fab/iGL-VRC01 Fab, iv9 Fab/iGL-VRC01 Fab and iv12 Fab/iGL-12A21Fab were concentrated to ~4.5, ~9, ~10, and ~10 mg/mL, respectively (Table S3). Complexes were screened using commercially available screens: Rigaku Wizard Precipitant Synergy block #2, Molecular Dimensions Proplex HT-96, Clear Strategy screens #1, and Hampton Research Crystal Screen HT; using vapor diffusion method. An NT8 dispensing robot, and Rock Imager was used to set up and screen crystallization conditions. Crystallization conditions were as follows: 0.1 M HEPES pH 7.0, PEG 8000 for iv1 scFv/iGL-VRC01 Fab; 0.1 M Tris pH 7.5, 8, %w/v PEG 20K, 8%w/v PEG MME 550, 0.2 M KSCN for iv4-HL Fab/iGL-VRC01 Fab; 0.08% Sodium Acetate pH 3.5, 24% PEG 4000, 0.16M Ammonium Acetate for iv9 Fab/iGL-VRC01 Fab and 0.1M KCL, 0.1 M Tris pH8, and 15%w/v PEG MME 2 K for iv12 Fab/iGL-12A21 Fab.
Seydoux E., Wan Y.H., Feng J., Wall A., Aljedani S., Homad L.J., MacCamy A.J., Weidle C., Gray M.D., Brumage L., Taylor J.J., Pancera M., Stamatatos L, & McGuire A.T. (2021). Development of a VRC01-class germline targeting immunogen derived from anti-idiotypic antibodies. Cell reports, 35(5), 109084.
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2

Obtaining High-Resolution Crystals of AMMO1 Fab

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Crystals of AMMO1 Fab were obtained using a NT8 dispensing robot and screening was done with Rigaku Wizard Precipitant Synergy block#2, Molecular Dimensions Proplex screen HT-96, Hampton Research Crystal Screen HT by the vapor diffusion method. Crystals used for diffraction data were grown in 16.75% PEG 400, 13.4% PEG 3350, 0.1M MgCl2, 0.1M Tris pH 8.5. Crystals were cryo-protected in solutions containing 30% molar excess of their original reagents and 20% Glycerol. Crystal diffracted to 1.6 Å (See Table S4). Data was collected at ALS 5.1 and 5.2 and processed using HKL2000 (Otwinowski and Minor, 1997 ).
Snijder J., Ortego M.S., Weidle C., Stuart A.B., Gray M.D., McElrath M.J., Pancera M., Veesler D, & McGuire A.T. (2018). An antibody targeting the fusion machinery neutralizes dual-tropic infection and defines a site of vulnerability on Epstein-Barr virus. Immunity, 48(4), 799-811.e9.
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3

Protein Complex Crystallization Conditions

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Complexes of iv1 scFv/iGL-VRC01 Fab, iv4-HL Fab/iGL-VRC01 Fab, iv9 Fab/iGL-VRC01 Fab and iv12 Fab/iGL-12A21Fab were concentrated to ~4.5, ~9, ~10, and ~10 mg/mL, respectively (Table S3). Complexes were screened using commercially available screens: Rigaku Wizard Precipitant Synergy block #2, Molecular Dimensions Proplex HT-96, Clear Strategy screens #1, and Hampton Research Crystal Screen HT; using vapor diffusion method. An NT8 dispensing robot, and Rock Imager was used to set up and screen crystallization conditions. Crystallization conditions were as follows: 0.1 M HEPES pH 7.0, PEG 8000 for iv1 scFv/iGL-VRC01 Fab; 0.1 M Tris pH 7.5, 8, %w/v PEG 20K, 8%w/v PEG MME 550, 0.2 M KSCN for iv4-HL Fab/iGL-VRC01 Fab; 0.08% Sodium Acetate pH 3.5, 24% PEG 4000, 0.16M Ammonium Acetate for iv9 Fab/iGL-VRC01 Fab and 0.1M KCL, 0.1 M Tris pH8, and 15%w/v PEG MME 2 K for iv12 Fab/iGL-12A21 Fab.
Seydoux E., Wan Y.H., Feng J., Wall A., Aljedani S., Homad L.J., MacCamy A.J., Weidle C., Gray M.D., Brumage L., Taylor J.J., Pancera M., Stamatatos L, & McGuire A.T. (2021). Development of a VRC01-class germline targeting immunogen derived from anti-idiotypic antibodies. Cell reports, 35(5), 109084.
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