Storing and Handling Lyophilized Tubulin

Overview


Lyophilized Tubulin is supplied as an alternative to our cryopreserved Cycled Tubulin (Cat. No. 032005) for its ease of storage and handling. Lyophilized Tubulin is supplied as a white powder with a tubulin-to-powder weight ratio of 1:3. When reconstituted with ultrapure water to 40 mg/ml, the buffer conditions are 10 mM Sodium Phosphate, 0.5 mM MgCl2, 0.1 mM GTP, and 0.25 M Trehalose, pH 7.0.

Recommended Products


Protocol


1. Store

Store tubulin in a cool, dry environment.

2. Resuspend

Resuspend tubulin to 40 mg/ml in ice-cold Tubulin PEM Buffer (Cat. No.032002).

 NOTE :

1 mg tubulin = 3 mg powder

3. Incubate

Incubate the resuspended tubulin on ice for 15 minutes.

4. Clarify

Clarify the resuspended tubulin to remove any protein aggregates. Spin at 14,000 rpm for 1 minute at 4°C in a microcentrifuge.

5. Aliquot

Aliquot tubulin into experimental-sized batches to avoid repeated freeze-thaw cycles.

6. Freeze in Liquid Nitrogen

Freeze experimental-sized aliquots in liquid Nitrogen.

 NOTE :

Failure to flash freeze with liquid Nitrogen will result in significant reduction of activity.

7. Store at -80°C

Transfer frozen experimental aliquots to -80°C. When ready to use, thaw in 37°C water bath followed by immediate placement on ice.

 NOTE :

Avoid repeated freeze-thaw cycles. Discard any unused portion of the experimental aliquots.

Recommended Products


Protocol


Step 1

1. Store

Store tubulin in a cool, dry environment.

Step 2

2. Resuspend

Resuspend tubulin to 40 mg/ml in ice-cold Tubulin PEM Buffer (Cat. No.032002).

 NOTE :

1 mg tubulin = 3 mg powder

Step 3

3. Incubate

Incubate the resuspended tubulin on ice for 15 minutes.

Step 4

4. Clarify

Clarify the resuspended tubulin to remove any protein aggregates. Spin at 14,000 rpm for 1 minute at 4°C in a microcentrifuge.

Step 5

5. Aliquot

Aliquot tubulin into experimental-sized batches to avoid repeated freeze-thaw cycles.

Step 6

6. Freeze in Liquid Nitrogen

Freeze experimental-sized aliquots in liquid Nitrogen.

 NOTE :

Failure to flash freeze with liquid Nitrogen will result in significant reduction of activity.

Step 7

7. Store at -80°C

Transfer frozen experimental aliquots to -80°C. When ready to use, thaw in 37°C water bath followed by immediate placement on ice.

 NOTE :

Avoid repeated freeze-thaw cycles. Discard any unused portion of the experimental aliquots.

Overview


Lyophilized Tubulin is supplied as an alternative to our cryopreserved Cycled Tubulin (Cat. No. 032005) for its ease of storage and handling. Lyophilized Tubulin is supplied as a white powder with a tubulin-to-powder weight ratio of 1:3. When reconstituted with ultrapure water to 40 mg/ml, the buffer conditions are 10 mM Sodium Phosphate, 0.5 mM MgCl2, 0.1 mM GTP, and 0.25 M Trehalose, pH 7.0.

Recommended Products

Protocol


Step 1

1. Store

Store tubulin in a cool, dry environment.

Step 2

2. Resuspend

Resuspend tubulin to 40 mg/ml in ice-cold Tubulin PEM Buffer (Cat. No.032002).

 NOTE :

1 mg tubulin = 3 mg powder

Step 3

3. Incubate

Incubate the resuspended tubulin on ice for 15 minutes.

Step 4

4. Clarify

Clarify the resuspended tubulin to remove any protein aggregates. Spin at 14,000 rpm for 1 minute at 4°C in a microcentrifuge.

Step 5

5. Aliquot

Aliquot tubulin into experimental-sized batches to avoid repeated freeze-thaw cycles.

Step 6

6. Freeze in Liquid Nitrogen

Freeze experimental-sized aliquots in liquid Nitrogen.

 NOTE :

Failure to flash freeze with liquid Nitrogen will result in significant reduction of activity.

Step 7

7. Store at -80°C

Transfer frozen experimental aliquots to -80°C. When ready to use, thaw in 37°C water bath followed by immediate placement on ice.

 NOTE :

Avoid repeated freeze-thaw cycles. Discard any unused portion of the experimental aliquots.