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X-Ray Crystallography

Membrane Protein Kit

Crystallization Kits | Crystallization Plates | Optimization Reagents | Renaturation Kit


Membrane Protein Kit


The Basic Crystallization Kit for Membrane Proteins is a rapid, empirical, and efficient screening method to determine the best conditions for crystallization of hydrophobic macromolecules (primarily membrane proteins) in an empirical way. Based on known or published data for various proteins, broad ranges of buffers, pH, and precipitants in combination with a micelle-forming detergent are explored using the sparse matrix method with a minimal amount of proteins.
Membrane Protein Kit

The crystallization principle of the kit is based on the screening of the hydrophilic, charged parts of the macromolecules by the binding of the detergent to hydrophobic areas of the macromolecules and subsequent reduction of their solubility. Detergent mediated contacts then lead to precipitation or crystallization of the screening molecules.


Product Code Product Name Size
73513 Membrane Protein Kit 1 kit

Crystallization Basic Kit for Membrane Proteins Datasheet (139 kb PDF)

Features & Benefits:

  • Comprehensive, efficient screening. A single test series using sparse matrix methods and a minimal amount of sample covers the full range of known, published crystallization conditions.
  • Prefabricated reagents, flow charts, and observation sheets. Allows increased productivity while reducing laboratory time and efforts.
  • Extremely high-purity reagents. Reduces sample denaturing, ensures reproducibility, and allows flexible comparison of different detergent effects.

 

Flow Diagram for Membrane Protein Kit


Screening Parameters Basic Kit
pH Range 4.6 to 8.5
Buffers Tris, citrate, ADA, HEPES, acetate
Precipitating Salts* sodium and potassium tartrate; sodium, potassium and ammonium phosphate; sodium and magnesium chloride; sodium and zinc acetate; sodium citrate; lithium, magnesium and ammonium sulfate
Precipitating Organic Solvents* MPD, 2-propanol
PEGs* PEG 400, 4000, and 6000
Detergent** stock solution of choice
* As sole precipitant and/or as combinations
** Proceed from detergents that form small micelles to those with larger micelle sizes.
  Helpful Notes:
We recommend using detergents that form small micelles, followed by use of detergents which form increasingly larger micelles only as needed.

The ternary system of sample, detergent, and precipitant is a delicate system where small parameter changes can dramatically change interactions between sample-sample, sample-solvent, sample-detergent and detergent-detergent. Corresponding effects on macromolecular solubility and crystallization should be expected.

Temperature is an important variable in all detergent-driven crystallization experiments.

Streak-seeding can produce larger crystals from microcrystalline precipitates.

This kit consists of 50 ready-to-use solutions reflecting different crystallization conditions. When working with standard multiwell plates you may prefer to use just 48 different conditions. In this case we recommend omitting reagents 43 and 47, or two reagents of your choice.

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