窝窝午夜理论片影院,国产黄A片免费网站免费

BOUTIQUEResouces

A Complete Beginner's Guide to Protein Crystallization Research: Easily Embark on a Journey of Life Science Exploration

Protein crystallization is a critical method for investigating protein structure and function, and it plays a vital role in understanding the mechanisms of biomacromolecular interactions, disease pathogenesis, and drug development. It involves the process of protein molecules stacking regularly in a solution to form ordered crystals, which is influenced by multiple factors such as protein purity, concentration, and buffer composition. To obtain high-quality crystals, steps including sample preparation, condition screening, and optimization are required. Research on protein crystallization drives advancements in the fields of biology and medicine, and provides support for drug design and protein engineering. This article outlines the key points of protein crystallization, including preparation, methods, screening, and optimization, to serve as a reference for related research.

1. The History of Protein Crystallization Development

Research on protein crystallization began in the mid-19th century, when Hunefeld first reported hemoglobin crystals. In the 20th century, crystallization technology became an important method for protein purification and research—notably, John Jacob Abel’s crystallization of insulin and James B. Sumner’s confirmation that enzymes are crystalline proteins. In the late 1930s, X-ray analysis was first applied to protein crystals, opening a new chapter in structural biology. With the development of genetics and molecular biology, especially the application of DNA technology, research on protein crystallization has undergone significant acceleration and transformation since the 1980s.

2. Protein Crystallization: A Combination of Art and Science

Despite the convenience brought by advancements in modern molecular biology and crystallography for many experiments, protein crystallization remains a challenge that is both simple and complex. This process combines scientific methods with artistic intuition and creativity, while also being highly dependent on the accumulation of experience.

Since biomacromolecules (e.g., proteins) are composed of a large number of different atoms arranged in complex ways, the crystallization process involves numerous variables and influencing factors. These factors include protein purity, concentration, pH value, and ionic strength—each of which can have a significant impact on crystallization results (Tables 1 and 2).

Table 1. Chemical Variables Affecting Protein Crystal Growth
Purity of the sample	Genetic modification
Conformational flexibility of the sample	Molecular symmetry
Homogeneity of the sample	Stability and denaturation degree of the sample
pH value and buffer	Isoelectric point
Type and concentration of precipitant	Presence or absence of His-tags and other purification tags
Concentration of the sample	Thermal stability
Additives, cofactors, ligands, inhibitors, effectors, and excipients	pH stability
Chaotropes (e.g., urea, guanidine hydrochloride, ammonium sulfate)	Historical processes of sample acquisition, handling, and storage
Detergents	Proteolysis
Metal ions	Microbial contamination
Ionic strength	Sample preservation
Reducing agents or oxidizing agents	Sample handling and associated cleanliness
Source of the sample	Types and concentrations of anions and cations
Amorphous particulate matter	Relative supersaturation
Post-translational modifications	Initial and final concentrations of other reagents
Chemical modifications	Equilibration path and rate
Table 2. Physical Variables That May or Do Affect Protein Crystal Growth
Temperature		Electromagnetic field
Equilibration rate		Surface of the crystallization device
Crystallization method		Viscosity of reagents
Gravity, convection, and sedimentation		Heterogeneous and epitaxial nucleants
Vibration and sound		Geometry of the crystallization device
Volumes of sample and reagents		Time
Pressure		Dielectric properties of reagents

Crystallization experiments typically start with screening, which aims to identify the key variable combinations that can produce crystals. However, to obtain crystals with specific properties (e.g., for structural biology, purification, or biotherapeutic research), further optimization experiments are often required (Tables 3 and 4).

Table 3. Crystallization Methods – Achievement of Supersaturation
Vapor Diffusion Method (Sitting, Hanging, Sandwich)		Sequential Extraction (a data processing and analysis technique)
Batch Crystallization Method (Microbatch Crystallization)		pH Induction (simple, controllable)
Dialysis Technique (Microdialysis Technique plays an auxiliary role)		Temperature Induction (easy to operate, low cost; needs to be used in combination with other methods)
Free Interface Diffusion (Counter-Diffusion, Liquid Bridge)		Effector Addition (also referred to as "Silver Bullet")
Controlled Evaporation
Table 4. Reagents Used in Protein Crystallization
Salts (Ammonium Sulfate, Sodium Formate, Ammonium Phosphate, etc.)	Non-Volatile Organic Compounds ((±)-2-Methyl-2,4-Pentanediol, 1,6-Hexanediol, Glycerol, etc.)
Polymers (Polyethylene Glycol [molecular weight range: 200–20,000], Ethylene Imine, Jeffamine®, etc.)	Buffers (HEPES, Tris, Sodium Acetate, MES, etc.)
Volatile Organic Compounds (2-Propanol, 1,4-Dioxane, Ethanol, etc.)	Additives (Calcium Chloride, Sodium Chloride, Tris(2-carboxyethyl)phosphine, n-Octyl-β-D-glucoside, etc.)

Although modern technology provides abundant resources and tools, crystallization is not merely a matter of using these tools. A successful crystallization strategy needs to be based on certain fundamental principles. As proposed by pioneers in the field of crystallization such as Alex McPherson, considering these principles before conducting experiments will lay a solid foundation for your crystallization work.

3. Key Elements of Protein Crystallization

· Sample: The purity of the sample is the cornerstone of successful crystallization, and careful handling is required to ensure high homogeneity of the sample.

· Homogeneity: The consistency of the chemical structure and molecular weight of the protein sample is crucial for the formation of ordered crystals.

· Solubility: Optimize the dissolution conditions of the sample in the buffer, pursue monodispersity, and avoid aggregation and precipitation.

· Stability: Ensure that the protein maintains structural and functional stability during crystallization and prevent denaturation.

· Supersaturation: Induce the ordered arrangement of protein molecules to form crystal nuclei by controlling supersaturation.

· Association: Regulate the ordered interactions between protein molecules to promote ordered aggregation and avoid non-specific aggregation.

· Nucleation: Control the formation of initial crystallization nuclei of proteins in the solution to optimize the number, size, and quality of crystals.

· Diversity: Try a variety of methods and conditions to identify suitable crystallization conditions for specific proteins.

· Control: Precisely regulate experimental conditions and processes to ensure the accuracy and reproducibility of experimental results.

· Impurities: Maintain sample purity and reduce the interference of impurities on crystallization and subsequent research.

· Preservation: Properly protect the crystallized protein crystals to prevent damage and ensure the reliability of subsequent research.

Hampton Research (USA), whose products are distributed by Beijing XMJ Technology Co., Ltd., is a leading manufacturer specializing in protein crystallization research. Its extensive product portfolio covers a full range of products, from initial crystallization screening and crystallization optimization screening to custom single-component crystallization reagents and crystallization growth optimization reagents. These products can meet the needs of different protein crystallization research, including the crystallization of general proteins, membrane proteins, and protein complexes.

With its cutting-edge biomacromolecular crystallization technology and practical, diverse product line, Hampton Research (distributed by XMJ) provides comprehensive crystallization research reagents and laboratory consumables to crystal researchers worldwide. These products not only greatly improve the work efficiency of researchers but also help them achieve outstanding scientific research results. Hampton Research has become one of the most trusted brands in the field of crystallization research.

Best-Selling Protein Crystallization Products of Hampton Research (Distributed by XMJ)

Hampton Research Protein Crystallization Screening Kits

Brand	Item Number	Product Name	Specification	Introduction
Hampton	HR2-144	Index	10 ml, tube format	Classic Initial Crystallization Screening Kit for multiple reagent systems (including proteins, complexes, peptides, nucleic acids, and water-soluble small molecules), containing 96 conditions
Hampton	HR2-110	Crystal Screen	10 ml, tube format	Initial Crystallization Screening Kit for multiple reagent systems (including proteins, soluble peptides, nucleic acids, and water-soluble small molecules), containing 50 conditions
Hampton	HR2-112	Crystal Screen 2	10 ml, tube format	Initial Crystallization Screening Kit for multiple reagent systems (including proteins, soluble peptides, nucleic acids, and water-soluble small molecules), containing 48 conditions
Hampton	HR2-114	MembFac	10 ml, tube format	Initial Crystallization Screening Kit for membrane proteins, containing 48 conditions
Hampton	HR2-116	Natrix	10 ml, tube format	Initial Crystallization Screening Kit for nucleic acids and nucleic acid/protein complexes, containing 48 conditions
Hampton	HR2-117	Natrix 2	10 ml, tube format	Initial Crystallization Screening Kit for nucleic acids and nucleic acid/protein complexes, containing 48 conditions
Hampton	HR2-082	PEGRx 1	10 ml, tube format	Polymer and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions
Hampton	HR2-084	PEGRx 2	10 ml, tube format	Polymer and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions條件
Hampton	HR2-126	PEG/Ion Screen	10 ml, tube format	Polymer, Salt and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions
Hampton	HR2-098	PEG/Ion 2 Screen	10 ml, tube format	Polymer, Salt and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions
Hampton	HR2-107	SaltRx 1	10 ml, tube format	Salt and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions
Hampton	HR2-109	SaltRx 2	10 ml, tube format	Salt and pH Condition Initial Screening and Secondary Screening Kit, containing 48 conditions

In addition to the aforementioned products, XMJ also provides a variety of crystallization condition screening kits, including GRAS Screen?, the Grid Screen series, Low Ionic Strength Screen, XP Screen, JBScreen Wizard, and more. For inquiries and orders, please feel free to send email to info@xmjsci.com

Hampton Research Protein Crystallization Optimization Kit.

Brand	Item Number	Product Name	Specification	Introduction
Hampton	HR2-072	Solubility & Stability Screen	0.5 ml, Deep Well block format	Solubility and Stability Screening, Including 96 Conditions
Hampton	HR2-413	Solubility & Stability Screen 2	0.5 ml, Deep Well block format	Solubility and Stability Screening, Including 96 Conditions
Hampton	HR2-070	Slice pH	0.5 ml, Deep Well block format	pH Screening Kit, Including 96 Conditions
Hampton	HR2-138	Additive Screen	1 ml, Deep Well block format	Additive Screening Kit, Including 96 Conditions
Hampton	HR2-459	GRAS Additive	1 ml, Deep Well block format	GRAS Additive Screening, Including 96 Conditions
Hampton	HR2-096	Silver Bullets	0.50 ml, Deep Well block format	Additive Screening Kit, Including 96 Conditions
Hampton	HR2-407	Detergent Screen	0.25 ml, Deep Well block format	Additive Screening Kit, Including 96 Conditions
Hampton	HR2-214	Ionic Liquid Screen	0.5 ml, tube format	Ionic Liquid Screening Kit, Including 24 Conditions
Hampton	HR2-241	StockOptions pH	10 ml, tube format	Crystal Growth Screening Kit, Including 45 Conditions
Hampton	HR2-235	StockOptions Sodium Citrate	10 ml, tube format	Crystal Growth Screening Kit, Including 24 Conditions
Hampton	HR2-239	StockOptions Sodium Cacodylate	10 ml, tube format	Crystal Growth Screening Kit, Including 24 Conditions
Hampton	HR2-243	StockOptions MES	10 ml, tube format	Crystal Growth Screening Kit, Including 20 Conditions
Hampton	HR2-103	StockOptions Bis-Tris Propane	10 ml, tube format	Crystal Growth Screening Kit, Including 33 Conditions
Hampton	HR2-102	StockOptions HEPES	10 ml, tube format	Crystal Growth Screening Kit, Including 15 Conditions
Hampton	HR2-227	StockOptions Polymer	10 ml, tube format	Crystal Growth Screening Kit, Including 23 Conditions
Hampton	HR2-245	StockOptions Salt	10 ml, tube format	Crystal Growth Screening Kit, Including 49 Conditions
Hampton	HR2-073	CryoPro	1 ml, tube format	Crystal Cryoprotectant Screening, Including 48 Conditions
Hampton	HR2-501	50% v/v Jeffamine^® M-600^® pH 7.0	200 ml	Crystalline grade Jeffamine M-600 is used for the optimization of crystallization conditions.
Hampton	HR2-525	50% w/v Polyethylene glycol 1,500	200 mL	Crystalline grade PEG 1500 is used for the optimization of crystallization conditions.
Hampton	HR2-527	50% w/v Polyethylene glycol 3,350 Monodisperse	200 mL	Crystalline grade PEG 3350 is used for the optimization of crystallization conditions.
Hampton	HR2-529	50% w/v Polyethylene glycol 4,000	200 mL	Crystalline grade PEG 4000 is used for the optimization of crystallization conditions.

The single-component conditions included in the aforementioned kits can be ordered individually. In addition to the kits mentioned above, Ximeijie also provides a variety of crystallization optimization screening kits and reagents, including Proti-Ace?, Silica Hydrogel Kit, Heavy Atom Screens, I3C Phasing Kit, etc. For inquiries and orders, please feel free to send email to info@xmjsci.com.

Hampton Research Protein Crystallization Consumables and Tools

Brand	Item Number	Product Name	Specification	Introduction
Hampton	HR2-320	Seed Bead Kit	24 tubes with PTFE Seed Bead	Seed Preparation Beads
Hampton	HR3-105	MASTERBLOCK® 96 Deep Well polypropylene plate	50 plate case	96-Well Deep-Well Plate
Hampton	HR3-081	72 Well Microbatch Plate, Greiner 654102	untreated, hydrophobic - 270 plate case	72-Well Under-Oil Crystallization Plate
Hampton	HR3-125	Swissci 3 Well Midi Crystallization Plate in UVP (40)	40 plate case	3-Microdrop/96-Well Hanging Drop Plate
Hampton	HR3-158	Cryschem Plate	24 plate case	24-Well Hanging Drop Plate
Hampton	HR3-231	22 mm x 0.22 mm Siliconized circle cover slides	1.0 ounce pack (~120 slides)	Circular Silanized Glass Coverslips
Hampton	HR3-233	22 mm x 0.22 mm Siliconized circle cover slides	10.0 ounce case (~1,200 slides)	Circular Silanized Glass Coverslips
Hampton	HR3-609	Crystal Clear Sealing Film	100 pack	Crystallization Plate Sealing Film
Hampton	HR4-216	Crystal Crusher	5 pack	Crystal Crusher
Hampton	HR4-217	Crystal Probe	12 pack	Disposable Stainless Steel Crystal Needles
Hampton	HR4-733	CrystalCap	with Vial - 60 pack	Crystal Loop Base
Hampton	HR4-811	Micro-Tools Set	each	Crystal Handling Tool Set, including 8 crystal probes of different shapes

In addition to the aforementioned products, XMJ also provides various specifications of crystallization plates, loops, microbridges, sealing compounds, pucks, and other tool consumables, as well as complete toolkits. These products help researchers conduct experiments faster and more efficiently. For inquiries and orders, please feel free to send email to info@xmjsci.com

As the official authorized distributor of Hampton Research in China, XMJ is committed to providing users with excellent technical support and thoughtful after-sales service. XMJ has always adhered to its core philosophy of "We promise what we can do. We do what we have promised" to deliver high-quality services to users. If you want to learn more about product information, please c feel free to send email to info@xmjsci.com or visit the official website at m.whuzsb.com for inquiries and more details.

References and Readings

1.Der Chemismus in der tierescher Organization, Hunefeld, F 1. (1840)p.160.Leipzig University, Germany.

2.A brief h istory of protein crystal growth, MePherson, A. (1991). J. Cryst.Growth, 110,1-10.

3 über die Eiweisskorper verschiedenen Oelsamen, Ritthausen, H. (1880)Pfluegers Arch. 21,81-104.

4 The proteins of the Brazil nut, Osborne T.(1891). Am. Chem. J. 13,212-218.

5 Crystalline insulin., Abel, 」.J., Geiling, E. M. K., Roultier, O. A., Bell, F. M.&Wintersteiner, 0.(1927).]. Pharmacol. Exp. Ther. 31,65-85.

6 The Enzymes, Sumner, J.B.& Somers, G.F(1943). New York: Aca-demic Press.

7 Crystalline enzymes, Northrop, M., Kunitz, M. & Herriott, R. M. (1948)New York: Columbia University Press.

8 Present at the flood: How structural molecular biology came about,Dickerson,R.E.(2005).FASEB J.20,809-810.

9 Preparation and Analysis of Protein Crystals, McPherson, A.(1982)New York: lohn Wiley & Sons.

10.Current approaches to macromolecular crystallization. McPherson, A. (1990).Eur.Biochem.189,1-23.

11.Crystallization of Biological Macromolecules, McPherson, A.(1999)Cold Spring Harbor: Cold Spring Harbor Laboratory Press.

12Introduction to protein crystallization, McPherson, A. and Gavira, J.A.(2014) Acta Crystallographica F, Volume 70, Part 1, 2-20.

13.Some Words of Advice from an Old Hand, Alexander McPherson, pages1-9, in Protein Crystallization, Second Edition, Edited by Terese Berg-fors. International University Line, 2009.