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Crystallization Experiments  

Are you looking to grow or practice growing crystals? Try out some of these crystallization experiments.


Tetragonal Lysozyme Recipes
Lysozyme: 50 mg/ml in 0.1 M Sodium Acetate pH 4.6
Reagent: 8% w/v Sodium Chloride, 0.1 M Sodium Acetate pH 4.6
Additional Reagents: Index Reagent 8, 22, 28, 31, 34, 40, 58, 59, 69, 86, 88

Mix equal amounts of lysozyme with reagent, incubate at 4 or 22 degrees Celsius. Batch or vapor diffusion works fine.

Lysozyme: 50 mg/ml in 0.1 M Sodium Acetate pH 4.6
Reagent: 10% v/v Ethylammonium nitrate

Mix equal amounts of lysozyme with reagent, incubate at 4 or 22 degrees Celsius. Batch or vapor diffusion works fine.

Lysozyme: 50 mg/ml in 0.1 M Sodium Acetate pH 4.6
Reagent: 2.5 M Sodium Chloride

Mix equal amounts of lysozyme with reagent, incubate at 4 or 22 degrees Celsius. Batch or vapor diffusion works fine.
The 15 Minute Lysozyme Crystals - By Enrico Stura
Lysozyme: 100 mg/ml in 50 mM Sodium Acetate pH 4.5
Reagent: 30% w/v MPEG 5,000, 1.0 M Sodium Chloride, 50 mM Sodium Acetate pH 4.5

Mix equal amount of lysozyme with reagent. Dilute protein and/or MPEG and/or Sodium Chloride for less nucleation, larger, and better shaped crystals. Batch or vapor diffusion method works fine.
A Sweet Thaumatin Recipe
Thaumatin: 25 to 50 mg/ml in your choice of buffer (ADA, ACES, Bis-Tris) at about pH 6.5
Reagent: 1.5 M Potassium Sodium Tartrate or 1.5 M Sodium Tartrate
Additional Reagents: Index Reagent 26, 31, 78, 86

Mix equal amounts of Thaumatin with reagent and equilibrate vapor diffusion style over reagent.
Hampton's Glucose Isomerase
Glucose Isomerase: 20-30 mg/ml in water or 50 mM buffer pH 6 to 8
Reagent: Your choice or the following (1.5-2.5 M Ammonium Sulfate pH 6-9) or (10-15% w/v PEG 4000-8000, 0.2 M salt*, pH 6-9 0.6), or (1.0 M Sodium Citrate, pH 6-8), or (0.1-0.3 M Magnesium Formate), or (1.0 M Sodium Formate, pH 5-7), or (10-20% v/v MPD, 0.2 M salt*, pH 6-9), or (10-20% v/v PEG 400, 0.2 M salt*, pH 6-9), or 10-20% iso-Propanol, 0.2 M salt*, pH 6-9)
* = 0.2 M salt such as Ammonium Sulfate, Magnesium Chloride, Magnesium Acetate, Calcium Acetate, or Ammonium Acetate.

Mix equal amounts of glucose isomerase and reagent and equilibrate vapor diffusion style over reagent.
Alpha Lactalbumin from Cow Milk - By Alexander McP
Alpha Lactalbumin: 30 to 40 mg/ml in water or 10 mM Tris-HCl buffer pH 8.5
Reagent: 30% w/v PEG 3350, 0.10 M Tris HCl pH 8.5, 0.2 M Lithium Sulfate

Mix equal amounts of Alpha Lactalbumin and reagent and equilibrate vapor diffusion style over reagent.
Classic Catalase
Beef Liver Catalase: 30 mg/ml in 25 mM Hepes pH 7
Reagent: Crystal Screen 25, 36

Mix equal amounts of Catalase with reagent. Vapor diffusion works great.
Precious Ribonuclease S
Bovine Pancrease Ribonuclease S: 50 mg/ml in 25 mM Hepes pH 7.5
Reagent: Crystal Screen 9, 20

Mix equal amounts of Ribonuclease S with reagent. Vapor diffusion method.
Pounds of Proteinase K
Proteinase K: 20 mg/ml in 25 mM Hepes pH 7.0, PMSF
Reagent: Crystal Screen 2, 3, 4, 6, 7, 9, 10, 11, 15, 16, 17, 18, 20, 22, 25, 28, 29, 30, 31, 32, 33, 34, 35, 36, 38, 39, 40, 41, 42, 44, 45, 46, 47, 48, 49, 50

Mix equal amounts of Proteinase K with reagent. Vapor diffusion or microbatch works well. This stuff grows anytime, anywhere, in anything... almost.
Crystals for Cryo
Glucose Isomerase: 20 to 30 mg/ml in 25 mM Tris pH 7.5
Reagent: Crystal Screen Cryo Reagent 21

Mix equal amounts of Glucose Isomerase and reagent. Vapor diffusion method. Mount crystal in CryoLoop. Swish mounted crystal through reservoir to increase MPD concentration in crystal to decrease mosaicity. No swish = more mosaicity. Crystal can diffract to 1.6, indexes in I-centered orthorhombic with a=92.7 b=97.4 c=102.7.

Mosaicity is about 0.5 t0 0.6
Reagent: Crystal Screen Cryo Reagent 23

Mix equal amounts of Glucose Isomerase and reagent. Vapor diffusion method. Mount crystal in CryoLoop. Mosaicity may be a bit more in this reagent and the unit cell will shrink a tad.

Special thanks to Laurie Betts for the the cryo work and data!
Tricky Trypsin
Bovine Pancreas Trypsin: 60 mg/ml in 10 mM Calcium Chloride, 10 mg/ml Benzamidine Hydrochloride, 25 mM Hepes pH 7.0
Reagent: Crystal Screen 4, 15, 16, 20, 28, 30, 31

Mix equal amounts of Trypsin with reagent. Vapor diffusion method works nicely.
Non-Newtonian Fluid
And now for something completely different...

2 cups corn starch plus 1 cup water.

A non-Newtonian fluid is a fluid whose viscosity is variable based on applied stress. The most commonly known non-Newtonian fluid is cornstarch dissolved in water. Contrast with Newtonian fluids like water, whose behavior can be described exclusively by temperature and pressure, not the forces acting on it from second to second. Non-Newtonian fluids are fascinating substances that can be used to help us understand physics in more detail, in an exciting, hands-on way.

If you punch a bucket full of a shear thickening non-Newtonian fluid, the stress introduced by the incoming force causes the atoms in the fluid to rearrange such that it behaves like a solid. Your hand will not go through. If you shove your hand into the fluid slowly, however, it will penetrate successfully. If you pull your hand out abruptly, it will again behave like a solid, and you can literally pull a bucket of the fluid out of its container in this way.
Slime
And now for something completely different...

Slime is easy to make. It only takes a few ingredients and a few minutes to make a batch. Follow these step-by-step written instructions or watch the video to see how to make slime. To get started, gather the following materials:

* water
* white glue (something like Elmer's)
* borax
* food coloring (unless you want uncolored white slime)

Mix the glue, water, and food coloring separately from the borax and water.

After you have dissolved the borax and diluted the glue, you are ready to combine the two solutions. Stir one slime solution into the other. Your slime will begin to polymerize immediately.

The slime will become hard to stir after you mix the borax and glue solutions. Try to mix it up as much as you can, then remove it from the bowl and finish mixing it by hand. It's okay if there is some colored water remaining in the bowl.

The slime will start out as a highly flexible polymer. You can stretch it and watch it flow. As you work it more, the slime will become stiffer and more like putty. Then you can shape it and mold it, though it will lose its shape over time. Don't eat your slime and don't leave it on surfaces that could be stained by the food coloring.

Store your slime in a sealed ziplock bag, preferably in the refrigerator. Insect pests will leave slime alone because borax is a natural pesticide, but you'll want to chill the slime to prevent mold growth if you live in an area with high mold count. The main danger to your slime is evaporation, so keep it sealed when you're not using it.
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