PACT premier TM HT-96 MD1-36 PACT premier is a ph, Anion, Cation crystallization trial devised to test ph within a PEG/Ion screen.

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1 PACT premier TM HT-96 MD1-36 PACT premier is a ph, Anin, Catin crystallizatin trial devised t test ph within a PEG/In screen. MD1-36 cntains 96 x 1 ml reagents in a deep-well blck. Features f PACT premier A mdern, cmprehensive PEG/in screen. This 96-well screen is really 3 screens in ne: 24-well ph/peg screen 24-well catin/peg screen 48-well anin/peg screen Ratinale fr a new PEG/In screen The first step in crystallizatin is ften t reach fr a cmmercially available sparse matrix kit, and hpe that ne f the cnditins prduces smething that lks harvestable, r ptimizable. If n bvius leads cme ut f the screen, it is hard t learn anything frm the negative (precipitate and clear) results. Traditinal PEG/In screens prvide a lgical test f seven catins and eleven anins using PEG 3350 as the precipitatin agent. Hwever, the user has n cntrl ver ph and hence cannt determine the effect f ne catin r anin ver anther. Fr this reasn the PEG/ION/pH (PACT) screen has been develped t systematically test the effect f ph, anins and catins, using PEG as the precipitant. This screen has been implemented very successfully at the Netherlands Cancer Institute (NKI), and at the Oxfrd Prtein Prductin Facility (OPPF). PACT premier ph/peg screen This cnsists f fur brad range buffer systems (1) versus PEG These buffers allw ne t scan the ph range frm 4 t 9, withut changing the chemistry f the system, s effectively islating the effect f ph frm the effect f the buffer that causes the change in ph. There are a few screens that try t test crystallizatin space in a mre ratinal manner fr example, the Clear Strategy Screen and The Slubility Tl Kit. Prtein crystals grwn successfully with PACT premier. 1

2 PACT premier Catin/PEG screen This is made up f six catins (all with chlride cunter ins) that are cmbined with PEG 6000 at fur different phs: Acetate ph 5, MES ph 6, HEPES ph 7 and Tris ph 8. The catins tested are Na +, NH 4 +, Li +, Mg 2+, Ca 2+ and Zn 2+. The zinc in is tested at lwer cncentratin than the ther catins in the screen (0.01 M vs. 0.2 M) Anin/PEG screen This is made up f 12 anins, with either sdium r ptassium cunter ins, which are tested at 0.2 M against PEG The anins include fluride, brmine, idide, thicyanate, nitrate, frmate, acetate, sulfate, tartrate, phsphate, citrate and malnate. The phsphate slutin is tested at a cncentratin f 0.02 M. Chlride is nt included here as it is the cunter in in the catin screen. Three sets f reagents are tested at ph 6.5, 7.5, and 8.5 with the Bis-Tris-Prpane buffer system whilst ne set f reagents is tested withut buffering. Frmulatin Ntes PACT premier reagents are frmulated using ultrapure water (>18.0 MΩ) and are sterile-filtered using 0.22 µm filters. N preservatives are added. Final ph may vary frm that specified n the datasheet. Mlecular Dimensins will be happy t discuss the precise frmulatin f individual reagents. Individual reagents and stck slutins fr ptimizatin are available frm Mlecular Dimensins. Enquiries regarding PACT-premier frmulatin, interpretatin f results r ptimizatin strategies are welcme. Please , fax r phne yur query t Mlecular Dimensins. Cntact and prduct details can be fund at References: (1)Newman et al (2005). Twards ratinalizatin f crystallizatin screening fr small- t medium-sized academic labratries: the PACT/JCSG+ strategy. Acta Cryst. D61, PACT premier- when used tgether with JCSG plus as a primary screening strategy is an extremely pwerful and successful cmbinatin, (i.e. a cmbinatin f a mdern sparse matrix apprach and an infrmatin yielding systematic trial). 2

3 PACT premier HT-96 Cnditins A1 D12 MD1-36 Well # Cnc. Salt Cnc. Buffer ph Cnc. Precipitant A1 0.1 M SPG % w/v PEG 1500 A2 0.1 M SPG % w/v PEG 1500 A3 0.1 M SPG % w/v PEG 1500 A4 0.1 M SPG % w/v PEG 1500 A5 0.1 M SPG % w/v PEG 1500 A6 0.1 M SPG % w/v PEG 1500 A7 0.2 M Sdium chlride 0.1 M Sdium acetate % w/v PEG 6000 A8 0.2 M Ammnium chlride 0.1 M Sdium acetate % w/v PEG 6000 A9 0.2 M Lithium chlride 0.1 M Sdium acetate % w/v PEG 6000 A M Magnesium chlride hexahydrate 0.1 M Sdium acetate % w/v PEG 6000 A M Calcium chlride dihydrate 0.1 M Sdium acetate % w/v PEG 6000 A M Zinc chlride 0.1 M Sdium acetate % w/v PEG 6000 B1 0.1 M MIB % w/v PEG 1500 B2 0.1 M MIB % w/v PEG 1500 B3 0.1 M MIB % w/v PEG 1500 B4 0.1 M MIB % w/v PEG 1500 B5 0.1 M MIB % w/v PEG 1500 B6 0.1 M MIB % w/v PEG 1500 B7 0.2 M Sdium chlride 0.1 M MES % w/v PEG 6000 B8 0.2 M Ammnium chlride 0.1 M MES % w/v PEG 6000 B9 0.2 M Lithium chlride 0.1 M MES % w/v PEG 6000 B M Magnesium chlride hexahydrate 0.1 M MES % w/v PEG 6000 B M Calcium chlride dihydrate 0.1 M MES % w/v PEG 6000 B M Zinc chlride 0.1 M MES % w/v PEG 6000 C1 0.1 M PCTP % w/v PEG 1500 C2 0.1 M PCTP % w/v PEG 1500 C3 0.1 M PCTP % w/v PEG 1500 C4 0.1 M PCTP % w/v PEG 1500 C5 0.1 M PCTP % w/v PEG 1500 C6 0.1 M PCTP % w/v PEG 1500 C7 0.2 M Sdium chlride 0.1 M HEPES % w/v PEG 6000 C8 0.2 M Ammnium chlride 0.1 M HEPES % w/v PEG 6000 C9 0.2 M Lithium chlride 0.1 M HEPES % w/v PEG 6000 C M Magnesium chlride hexahydrate 0.1 M HEPES % w/v PEG 6000 C M Calcium chlride hexahydrate 0.1 M HEPES % w/v PEG 6000 C M Zinc chlride 0.1 M HEPES % w/v PEG 6000 D1 0.1 M MMT % w/v PEG 1500 D2 0.1 M MMT % w/v PEG 1500 D3 0.1 M MMT % w/v PEG 1500 D4 0.1 M MMT % w/v PEG 1500 D5 0.1 M MMT % w/v PEG 1500 D6 0.1 M MMT % w/v PEG 1500 D7 0.2 M Sdium chlride 0.1 M Tris % w/v PEG 6000 D8 0.2 M Ammnium chlride 0.1 M Tris % w/v PEG 6000 D9 0.2 M Lithium chlride 0.1 M Tris % w/v PEG 6000 D M Magnesium chlride hexahydrate 0.1 M Tris % w/v PEG 6000 D M Calcium chlride dihydrate 0.1 M Tris % w/v PEG 6000 D M Zinc chlride 0.1 M Tris % w/v PEG

4 PACT premier HT-96 Cnditins E1 H12 MD1-36 Well # Cnc. Salt Cnc. Buffer ph Cnc. Precipitant E1 0.2 M Sdium fluride 20 % w/v PEG 3350 E2 0.2 M Sdium brmide 20 % w/v PEG 3350 E3 0.2 M Sdium idide 20 % w/v PEG 3350 E4 0.2 M Ptassium thicyanate 20 % w/v PEG 3350 E5 0.2 M Sdium nitrate 20 % w/v PEG 3350 E6 0.2 M Sdium frmate 20 % w/v PEG 3350 E7 0.2 M Sdium acetate trihydrate 20 % w/v PEG 3350 E8 0.2 M Sdium sulfate 20 % w/v PEG 3350 E9 0.2 M Ptassium sdium tartrate tetrahydrate 20 % w/v PEG 3350 E M Sdium/ptassium phsphate 20 % w/v PEG 3350 E M Sdium citrate tribasic dihydrate 20 % w/v PEG 3350 E M Sdium malnate dibasic mnhydrate 20 % w/v PEG 3350 F1 0.2 M Sdium fluride 0.1 M Bis-Tris prpane % w/v PEG 3350 F2 0.2 M Sdium brmide 0.1 M Bis-Tris prpane % w/v PEG 3350 F3 0.2 M Sdium idide 0.1 M Bis-Tris prpane % w/v PEG 3350 F4 0.2 M Ptassium thicyanate 0.1 M Bis-Tris prpane % w/v PEG 3350 F5 0.2 M Sdium nitrate 0.1 M Bis-Tris prpane % w/v PEG 3350 F6 0.2 M Sdium frmate 0.1 M Bis-Tris prpane % w/v PEG 3350 F7 0.2 M Sdium acetate trihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 F8 0.2 M Sdium sulfate 0.1 M Bis-Tris prpane % w/v PEG 3350 F9 0.2 M Ptassium sdium tartrate tetrahydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 F M Sdium/ptassium phsphate 0.1 M Bis-Tris prpane % w/v PEG 3350 F M Sdium citrate tribasic dihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 F M Sdium malnate dibasic mnhydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 G1 0.2 M Sdium fluride 0.1 M Bis-Tris prpane % w/v PEG 3350 G2 0.2 M Sdium brmide 0.1 M Bis-Tris prpane % w/v PEG 3350 G3 0.2 M Sdium idide 0.1 M Bis-Tris prpane % w/v PEG 3350 G4 0.2 M Ptassium thicyanate 0.1 M Bis-Tris prpane % w/v PEG 3350 G5 0.2 M Sdium nitrate 0.1 M Bis-Tris prpane % w/v PEG 3350 G6 0.2 M Sdium frmate 0.1 M Bis-Tris prpane % w/v PEG 3350 G7 0.2 M Sdium acetate trihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 G8 0.2 M Sdium sulfate 0.1 M Bis-Tris prpane % w/v PEG 3350 G9 0.2 M Ptassium sdium tartrate tetrahydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 G M Sdium/ptassium phsphate 0.1 M Bis-Tris prpane % w/v PEG 3350 G M Sdium citrate tribasic dihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 G M Sdium malnate dibasic mnhydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 H1 0.2 M Sdium fluride 0.1 M Bis-Tris prpane % w/v PEG 3350 H2 0.2 M Sdium brmide 0.1 M Bis-Tris prpane % w/v PEG 3350 H3 0.2 M Sdium idide 0.1 M Bis-Tris prpane % w/v PEG 3350 H4 0.2 M Ptassium thicyanate 0.1 M Bis-Tris prpane % w/v PEG 3350 H5 0.2 M Sdium nitrate 0.1 M Bis-Tris prpane % w/v PEG 3350 H6 0.2 M Sdium frmate 0.1 M Bis-Tris prpane % w/v PEG 3350 H7 0.2 M Sdium acetate trihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 H8 0.2 M Sdium sulfate 0.1 M Bis-Tris prpane % w/v PEG 3350 H9 0.2 M Ptassium sdium tartrate tetrahydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 H M Sdium/ptassium phsphate 0.1 M Bis-Tris prpane % w/v PEG 3350 H M Sdium citrate tribasic dihydrate 0.1 M Bis-Tris prpane % w/v PEG 3350 H M Sdium malnate dibasic mnhydrate 0.1 M Bis-Tris prpane % w/v PEG

5 Abbreviatins: HEPES; N-(2-hydrxyethyl)-piperazine-N'-2-ethanesulfnic acid, MES; 2-(N-mrphlin)ethanesulfnic acid, PEG; Plyethylene glycl, Tris; 2-Amin-2-(hydrxymethyl)prpane-1,3-dil, SPG buffer; Succinic Acid, Phsphate, Glycine, MIB buffer; Malnic acid, Imidazle, Bric acid, PCTP buffer; Prpinic acid, Cacdylate, Bis-tris prpane, MMT buffer; Malic acid, MES, Tris. Manufacturer s safety data sheets are available frm ur website r by scanning the QR cde here: Ordering details: Catalgue Descriptin Catalgue Cde PACT premier (96 x 10 ml kit) MD1-29 PACT premier HT-96 (96 x 1 ml) MD1-36 PACT premier Ec Screen (96 x 10 ml kit) MD1-29-ECO PACT premier HT-96 Ec Screen (96 x 1 ml) MD1-36-ECO Green Screens PACT premier Green Screens (96 x 10 ml kit) MD1-55 PACT premier HT-96 Green Screens (96 x 1 ml) MD1-52 Single Reagents PACT premier (100 ml) MDSR-29 - tube number PACT premier HT-96 (100 ml) MDSR-36 - well number PACT premier Green screen (100 ml) MDSR-55 - tube number PACT premier Green screen HT-96 (100 ml) MDSR-52 - well number Fr PACT premier stck reagents visit ur Optimizatin page n ur website. 5

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