MD1-64-GREEN is a targeted sparse matrix presented as a 96 x 1 ml deep-well block.

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MemGold2 HT-96 Green Screen MD1-64-GREEN MemGold2 - The latest innovation for crystallization of membrane proteins. This screen targets all alpha helical types of Prokaryotic and Eukaryotic membrane proteins and with our non-covalent fluorescent dye added for superb UV performance. MD1-64-GREEN is a targeted sparse matrix presented as a 96 x 1 ml deep-well block. Features of MemGold2 Green Screen: Contains a non-covalent fluorescent dye (1,8- ANS) for enhanced UV performance. A brand new set of 96 of the most recent alphahelical membrane protein crystallization conditions. Particularly suited for Prokaryotic and Eukaryotic alpha-helical membrane proteins. Increased crystal contrast- observe protein crystals <30µM and also those lacking intrinsic fluorescence. A great addition to any membrane protein lab. Works with MemGold, MemStart, MemSys & MemPlus. Screening over a wider range of ph s (4-10). Addition of small MW PEGs. Can be used in conjunction with Lipidic Sponge Phase and/or Lipidic Cubic Phases. Introduction: In 2008 Molecular Dimensions released MemGold (1) - a rationalized sparse matrix type membrane protein crystallization screen. MemGold was based on the crystallization conditions for 121 alpha helical Membrane Proteins deposited in the PDB. Since MemGold, the number of structures has more than doubled. In response to this, MemGold2 (2) has been developed. MemGold2 includes a further 96 crystallization conditions from unique alpha helical Membrane Protein structures including channel and transporter structures, GPCRs and ATPases. It is suitable for both Prokaryotic and Eukaryotic alpha helical membrane proteins. Formulation Notes: MemGold2 reagents are formulated using ultrapure water (>18.0 MΩ) and are sterile-filtered using 0.22 µm filters. No preservatives are added. Final ph may vary from that specified on the datasheet. Molecular Dimensions will be happy to discuss the precise formulation of individual reagents. Individual reagents and stock solutions for optimization are available from Molecular Dimensions. Enquiries regarding MemGold2 formulation, interpretation of results or optimization strategies are welcome. Please e-mail, fax or phone your query to Molecular Dimensions. Contact and product details can be found at www.moleculardimensions.com *References: (1) Newstead, S., Ferrandon, S., and Iwata, S. Rationalizing alpha-helical membrane protein crystallization Volume 17, Issue 3, pages 466 472, March 2008 - Protein Science, 2008 - Wiley Online Library. (2) Parker, J. and Newstead, S. Current trends in alpha helical membrane protein crystallization: an update, Protein Science, 2012, 21(9):1358-65. (3) 2. Groves et al. A method for the general identification of protein crystals in crystallization experiments using a non-covalent fluorescent dye. Acta Cryst. 2007, D63, 526-535. Examples of membrane protein crystals grown using MemGold2 and the structure of a membrane transport protein (previously solved using MemGold ). 1

Why use UV fluorescence? In recent years, UV fluorescence imaging has become an established method for protein/salt crystal discrimination. Whilst the majority of proteins contain at least one tryptophan residue there remain a number that fluoresce weakly or not at all, yielding a false negative Green Screens contain a non-covalent fluorescent dye (3), (1,8-ANS, emission at 490 nm) conveys fluorescence on most proteins when illuminated with UV light. This not only increases the signal-to-noise ratio (important for very small crystals), but also allows the identification of protein crystals lacking intrinsic fluorescence that would otherwise remain ambiguous. Use these screens together with UV transparent plates (MD11-00U-100) and ClearVue2 sheets (MD6-19) for optimum UV performance. Features of the Green Screens: Non-covalent binds in channels and not observed to affect crystallization or diffraction quality in any of the proteins tested*. Increased crystal contrast observe protein crystals <30 μm and also those lacking significan intrinsic fluorescence. Improved signal-to-noise on average quantum yield ratio of fluorescence is increased from 0.2 to 0.7. Standard format for all automated systems. a c d b Images of a crystallization drop showing (a) crystals under skin with white light and (b) the same drop under UV- with 1,8-ANS dye and (c) A clear drop under white light? (d) the same drop showing a micro-crystal found using 1,8- ANS dye. Images kindly provided by Matthew Groves, EMBL Outstation-Hamburg. 2

MemGold2 HT-96 Green Screen Conditions A1 - D12 MD1-64-GREEN Well # Conc. Salt Conc. Buffer ph Conc. Precipitant Dye concentration A1 0.2 M Magnesium chloride hexahydrate 0.1 M Tris 7.5 14 % v/v PEG 500 MME 100µM 1,8-ANS 0.005 M Cadmium chloride hemi(pentahydrate) A2 0.1 M Potassium acetate 0.02 M Tris 7.0 44 % w/v PEG 3000 100µM 1,8-ANS 0.01 M Potassium chloride A3 0.08 M Magnesium sulfate heptahydrate 0.02 M MES 6.0 10 % w/v PEG 1450 100µM 1,8-ANS 0.02 M Sodium chloride A4 0.04 M Magnesium sulfate heptahydrate 0.02 M MES 6.5 8 % w/v PEG 1450 100µM 1,8-ANS 0.02 M Sodium chloride A5 0.05 M Sodium sulfate 0.05 M Tris 8.5 32 % v/v PEG 400 100µM 1,8-ANS 0.05 M Lithium chloride A6 0.1 M Sodium phosphate monobasic monohydrate 0.1 M Bis-Tris propane 7.5 10 % w/v PEG 3350 100µM 1,8-ANS 0.1 M Potassium phosphate dibasic A7 0.1 M Sodium chloride 0.1 M ADA 6.5 11.5 % w/v PEG 4000 100µM 1,8-ANS 0.1 M Lithium sulfate A8 0.1 M Lithium chloride 0.1 M Sodium acetate 4.5 30 % v/v PEG 400 100µM 1,8-ANS 0.1 M Cadmium chloride hemi(pentahydrate) A9 0.2 M Ammonium sulfate 0.1 M Sodium citrate 6.0 20 % w/v PEG 2000 100µM 1,8-ANS 0.1 M Sodium chloride A10 0.2 M Lithium sulfate 0.1 M HEPES 7.0 31 % v/v PEG 400 100µM 1,8-ANS 0.1 M Sodium chloride A11 0.2 M Ammonium phosphate monobasic 0.1 M Sodium citrate 4.5 32 % v/v PEG 400 100µM 1,8-ANS 0.1 M Ammonium sulfate A12 0.05 M Sodium citrate tribasic dihydrate 0.08 M Bis-Tris 6.0 14 % w/v PEG 4000 100µM 1,8-ANS 0.12 M Potassium chloride B1 0.1 M Sodium chloride 0.01 M MES 6.5 19 % w/v PEG 1000 100µM 1,8-ANS 0.15 M Ammonium sulfate B2 0.01 M Nickel(II) sulfate hexahydrate 0.1 M Sodium citrate 6.0 18 % w/v PEG 2000 MME 100µM 1,8-ANS B3 0.02 M Magnesium chloride hexahydrate 0.02 M MES 6.0 3.5 % w/v PEG 3350 100µM 1,8-ANS B4 0.02 M Sodium chloride 0.05 M MES 5.5 14 % v/v PEG 350 MME 100µM 1,8-ANS B5 0.025 M Magnesium chloride hexahydrate 0.02 M MOPS 7.0 35 % v/v PEG 500 MME 100µM 1,8-ANS B6 0.03 M Magnesium chloride hexahydrate 0.1 M MES 6.5 28 % v/v PEG 400 100µM 1,8-ANS B7 0.04 M Sodium chloride 0.04 M Tris 8.0 25 % v/v PEG 350 MME 100µM 1,8-ANS B8 0.04 M Magnesium acetate tetrahydrate 0.1 M MES 6.0 36 % v/v MPD 100µM 1,8-ANS B9 0.05 M Zinc acetate dihydrate 0.05 M ADA 6.3 11 % w/v PEG 8000 100µM 1,8-ANS B10 0.05 M Magnesium acetate tetrahydrate 0.1 M MES 6.5 26 % v/v PEG 400 100µM 1,8-ANS B11 0.05 M Magnesium acetate tetrahydrate 0.1 M Glycine 9.5 32 % v/v PEG 400 100µM 1,8-ANS B12 0.066 M Sodium chloride 0.02 M Tris 7.5 3 % w/v PEG 4000 100µM 1,8-ANS C1 0.075 M Magnesium chloride hexahydrate 0.1 M Sodium cacodylate 6.5 30 % w/v PEG 2000 MME 100µM 1,8-ANS C2 0.08 M Magnesium acetate tetrahydrate 0.1 M Sodium citrate 6.0 14 % w/v PEG 5000 MME 100µM 1,8-ANS C3 0.01 M Zinc acetate dihydrate 0.1 M MES 6.0 100µM 1,8-ANS 1.5 M Ammonium sulfate C4 0.087 M Ammonium sulfate 0.05 M Tris 7.0 22 % v/v PEG 250 DME 100µM 1,8-ANS C5 0.1 M Magnesium chloride hexahydrate 0.1 M Tris 7.5 13 % w/v PEG 8000 100µM 1,8-ANS C6 0.1 M Magnesium formate dihydrate 0.1 M MOPS 7.0 17 % w/v PEG 3350 100µM 1,8-ANS C7 0.1 M Potassium chloride 0.1 M Bis-Tris 6.0 18 % w/v PEG 4000 100µM 1,8-ANS C8 0.1 M Potassium chloride 0.1 M Potassium phosphate 7.5 18 % v/v PEG 200 100µM 1,8-ANS C9 0.1 M Magnesium acetate tetrahydrate 0.1 M MES 6.0 22 % w/v PEG 4000 100µM 1,8-ANS C10 0.1 M Calcium acetate hydrate 0.1 M MES 6.0 22 % w/v PEG 8000 100µM 1,8-ANS C11 0.1 M Ammonium sulfate 0.1 M HEPES 8.5 23 % w/v PEG 3350 100µM 1,8-ANS C12 0.1 M Potassium chloride 0.1 M MES 6.0 32 % v/v PEG 400 100µM 1,8-ANS D1 0.1 M Sodium chloride 0.1 M MES 6.5 36 % v/v PEG 300 100µM 1,8-ANS D2 0.1 M Sodium chloride 0.1 M BICINE 9.0 45 % v/v PEG 500 MME 100µM 1,8-ANS D3 0.15 M Calcium chloride dihydrate 0.1 M Glycine 9.0 35 % v/v PEG 400 100µM 1,8-ANS D4 0.2 M Ammonium sulfate 0.05 M ADA 6.5 13 % w/v PEG 4000 100µM 1,8-ANS D5 0.2 M Choline chloride 0.1 M Tris 7.5 14 % w/v PEG 2000 MME 100µM 1,8-ANS D6 0.2 M Sodium chloride 0.05 M MOPS 7.0 19 % w/v PEG 6000 100µM 1,8-ANS D7 0.05 M Sodium chloride 0.05 M MOPS 7.0 19 % w/v PEG 6000 100µM 1,8-ANS D8 0.2 M Magnesium formate dihydrate 0.05 M Tris 8.0 19 % w/v PEG 3350 100µM 1,8-ANS D9 0.2 M Calcium chloride dihydrate 0.1 M MES 5.0 20 % v/v PEG 350 MME 100µM 1,8-ANS D10 0.2 M Ammonium nitrate 0.05 M HEPES 7.0 20 % w/v PEG 3350 100µM 1,8-ANS D11 0.02 M Lithium chloride 0.02 M Glycine 10.0 33 % w/v PEG 1000 100µM 1,8-ANS 0.05 M Magnesium chloride hexahydrate D12 0.2 M Calcium acetate hydrate 0.1 M HEPES 7.0 24 % v/v PEG 400 100µM 1,8-ANS 3

MemGold2 HT96 Green Screen Conditions E1 - H12 MD1-64-GREEN Tube # Conc. Salt Conc. Buffer ph Conc. Precipitant Dye concentration E1 0.2 M Sodium acetate trihydrate 0.1 M MES 6.5 28 % v/v PEG 400 100µM 1,8-ANS E2 0.2 M Sodium chloride 0.05 M Calcium acetate 5.0 29 % v/v PEG 400 100µM 1,8-ANS E3 0.2 M Sodium chloride 0.1 M HEPES 7.0 29 % v/v PEG 400 100µM 1,8-ANS E4 0.2 M Ammonium formate 0.1 M Tris 7.0 31 % v/v Pentaerythritole ethoxylate (15/4 EO/OH) 100µM 1,8-ANS E5 0.2 M Ammonium sulfate 0.1 M Tris 8.5 35 % w/v PEG 3350 100µM 1,8-ANS E6 0.2 M Calcium acetate hydrate 0.1 M Sodium acetate 5.0 38 % v/v PEG 400 100µM 1,8-ANS E7 0.2 M Sodium chloride 0.1 M MOPS 7.5 38 % v/v PEG 400 100µM 1,8-ANS E8 2.0 M Ammonium sulfate 0.1 M Sodium cacodylate 6.5 100µM 1,8-ANS 0.2 M Sodium chloride E9 0.225 M Ammonium sulfate 0.05 M Sodium acetate 4.0 12 % w/v PEG 4000 100µM 1,8-ANS E10 0.23 M Sodium chloride 0.05 M Sodium acetate 4.5 33 % v/v PEG 400 100µM 1,8-ANS E11 0.25 M Magnesium formate dihydrate 0.1 M Sodium cacodylate 6.5 22 % w/v PEG 3000 100µM 1,8-ANS E12 0.25 M Magnesium chloride hexahydrate 0.1 M Tris 8.5 40 % w/v PEG 1000 100µM 1,8-ANS F1 0.3 M Lithium sulfate 0.1 M MES 6.5 25 % v/v PEG 400 100µM 1,8-ANS F2 0.3 M Ammonium formate 0.05 M Tris 9.0 33 % v/v PEG 500 MME 100µM 1,8-ANS F3 0.3 M Barium chloride dihydrate 0.1 M MES 6.0 34 % v/v PEG 400 100µM 1,8-ANS F4 0.32 M Lithium chloride 0.1 M Sodium citrate 5.5 14 % w/v PEG 4000 100µM 1,8-ANS F5 0.34 M Ammonium sulfate 0.1 M Sodium citrate 5.5 12 % w/v PEG 4000 100µM 1,8-ANS F6 0.35 M Lithium sulfate 0.1 M Sodium acetate 4.0 11 % v/v PEG 600 100µM 1,8-ANS F7 0.37 M Potassium nitrate 0.1 M MES 6.5 22 % v/v PEG 400 100µM 1,8-ANS F8 0.4 M Ammonium sulfate 0.1 M MES 6.5 10 % w/v PEG 3350 100µM 1,8-ANS F9 0.04 M Magnesium chloride hexahydrate 0.1 M HEPES 7.5 32 % v/v PEG 400 100µM 1,8-ANS 0.05 M Sodium chloride F10 0.4 M Potassium chloride 0.05 M HEPES 7.5 12 % v/v PEG 400 100µM 1,8-ANS F11 0.4 M Ammonium thiocyanate 0.1 M Sodium acetate 4.5 15 % w/v PEG 4000 100µM 1,8-ANS F12 0.4 M Sodium thiocyanate 0.1 M Sodium acetate 4.0 16 % w/v PEG 4000 100µM 1,8-ANS G1 0.5 M Potassium chloride 0.05 M HEPES 6.5 20 % v/v PEG 400 100µM 1,8-ANS G2 0.5 M Magnesium chloride hexahydrate 0.05 M Tris 7.5 21 % v/v PEG 350 MME 100µM 1,8-ANS G3 0.8 M Potassium formate 0.1 M Sodium acetate 5.0 11 % w/v PEG 4000 100µM 1,8-ANS G4 0.1 M MOPS 7.0 9 % w/v PEG 8000 100µM 1,8-ANS G5 0.1 M MES 6.0 11 % w/v PEG 20,000 100µM 1,8-ANS G6 0.1 M MES 6.5 13 % v/v PEG 400 100µM 1,8-ANS G7 0.1 M ADA 5.5 14 % w/v PEG 6000 100µM 1,8-ANS G8 0.05 M Tris 7.5 17 % v/v PEG 350 MME 100µM 1,8-ANS G9 0.07 M Sodium citrate 4.5 22 % v/v PEG 300 100µM 1,8-ANS G10 0.05 M ADA 6.5 24 % v/v PEG 400 100µM 1,8-ANS G11 0.1 M Sodium cacodylate 6.5 24 % w/v PEG 1500 100µM 1,8-ANS G12 0.1 M HEPES 7.5 28 % v/v PEG 600 100µM 1,8-ANS H1 0.05 M Tris 8.5 28 % v/v PEG 400 100µM 1,8-ANS H2 0.1 M BICINE 9.0 30 % v/v PEG 400 100µM 1,8-ANS H3 0.1 M ADA 7.0 31 % v/v PEG 600 100µM 1,8-ANS H4 0.1 M Tris 8.5 32 % v/v PEG 500 MME 100µM 1,8-ANS H5 0.1 M HEPES 7.5 33 % v/v PEG 400 100µM 1,8-ANS H6 0.18 M Sodium citrate 4.0 34 % w/v PEG 3350 100µM 1,8-ANS H7 0.1 M Tris 8.5 44 % v/v PEG 200 100µM 1,8-ANS H8 0.1 M Tris 8.0 65 % v/v MPD 100µM 1,8-ANS H9 2.75 M Ammonium chloride 0.025 M Bis-Tris 7.0 100µM 1,8-ANS H10 2.8 M Ammonium chloride 0.075 M HEPES 7.5 100µM 1,8-ANS H11 3.0 M Ammonium sulfate 0.1 M MES 5.5 100µM 1,8-ANS H12 0.01 M HEPES 7.5 3.25 M 1,6-Hexanediol 100µM 1,8-ANS Abbreviations: ADA; N-(2-Acetamido)iminodiacetic Acid, BICINE; N,N-Bis(2-hydroxyethyl)glycine, Bis-Tris; 2,2'-(Propane-1,3- diyldiimino)bis[2-(hydroxymethyl)propane-1,3-diol]. CHES; 2-(N-Cyclohexylamino)ethane sulfonic Acid, HEPES; N-(2- hydroxyethyl)-piperazine-n'-2-ethanesulfonic acid, MES; 2-(N-morpholino)ethanesulfonic acid, MME; Monomethylether, MOPS; 3-morpholinopropane-1-sulfonic acid, PEG; Polyethylene glycol, PEG DME; Poly(ethylene glycol) bis(carboxymethyl) ether, Tricine; N-[Tris(hydroxymethyl)methyl]glycine, Tris; 2-Amino-2-(hydroxymethyl)propane-1,3- diol, 1,8-ANS; 8-anilino-1-napthalenesulfonic acid 4

Manufacturer s safety data sheets are available from our website or by scanning the QR code here: Re-Ordering details: Catalogue Description Pack size Catalogue Code MemGold2 96 x 10 ml MD1-63 MemGold2 HT-96 96 x 1 ml MD1-64 Eco Screens MemGold2 Eco Screen 96 x 10 ml MD1-63-ECO MemGold2 HT-96 Eco Screen 96 x 1 ml MD1-64-ECO Green Screens (contain fluorescent green dye - ideal for UV) MemGold2 Green Screen 96 x 10 ml MD1-63-GREEN MemGold2 HT-96 Green Screen 96 x 1 ml MD1-64-GREEN Combo Packs MemGold Combo Value Pack 2 x 96 x 10 ml MD1-74 (MemGold + MemGold2 ) MemGold HT-96 Combo Value Pack 2 x 96 x 1 ml MD1-74-HT (MemGold + MemGold2 ) Single Reagents MemGold2 single reagents 100 ml MDSR-63-tube number MemGold2 HT-96 single reagents 100 ml MDSR-64-well number For MemGold2 stock solutions please visit the Optimization section on our website. 5

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