Storing and Handling Pure Actin

Overview


Pure Actin is supplied as a white powder with an actin-to-powder weight ratio of 1:3. When reconstituted with ultrapure water to 9 mg/ml, the buffer conditions are 2 mM Tris-HCl, 0.2 mM CaCl2 , 0.2 mM ATP, 1 mM DTT, and 0.25 M Trehalose, pH 8.0. Actin is unstable in the absence of ATP, a divalent cation, and DTT.

Recommended Products


Recommended Products


Overview


Pure Actin is supplied as a white powder with an actin-to-powder weight ratio of 1:3. When reconstituted with ultrapure water to 9 mg/ml, the buffer conditions are 2 mM Tris-HCl, 0.2 mM CaCl2 , 0.2 mM ATP, 1 mM DTT, and 0.25 M Trehalose, pH 8.0. Actin is unstable in the absence of ATP, a divalent cation, and DTT.

Recommended Products


Unlabeled Tubulin

Protocol


1. Store

Store actin in a cool, dry environment.

2. Resuspend

Resuspend actin to 9 mg/ml in ice-cold ultrapure water.

 NOTE :

1 mg actin = 3 mg powder

3. Dilute

Dilute to 0.4 mg/ml with Actin Working Buffer (Cat. No. 000102). Add ATP to 0.2 mM and DTT to 0.5 mM.

4. Incubate

Incubate the reconstituted actin on ice for 1 hour.

5. Clarify

Clarify the reconstituted actin to remove any aggregates. Spin at 14,000 rpm for 15 minutes at 4°C in a microcentrifuge.

5. Store

Add sodium azide to 0.05% and store at 4°C.

 NOTE :

The product is stable under these storage conditions for several weeks.

Protocol


Step 1

1. Store

Store actin in a cool, dry environment.

Step 2

2. Resuspend

Resuspend actin to 9 mg/ml in ice-cold ultrapure water.

 NOTE :

1 mg actin = 3 mg powder

Step 3

3. Dilute

Dilute to 0.4 mg/ml with Actin Working Buffer (Cat. No. 000102). Add ATP to 0.2 mM and DTT to 0.5 mM.

Step 4

4. Incubate

Incubate the reconstituted actin on ice for 1 hour.

Step 5

5. Clarify

Clarify the reconstituted actin to remove any aggregates. Spin at 14,000 rpm for 15 minutes at 4°C in a microcentrifuge.

Step 6

6. Store

Add sodium azide to 0.05% and store at 4°C.

 NOTE :

The product is stable under these storage conditions for several weeks.

Protocol


Step 1

1. Store

Store actin in a cool, dry environment.

Step 2

2. Resuspend

Resuspend actin to 9 mg/ml in ice-cold ultrapure water.

 NOTE :

1 mg actin = 3 mg powder

Step 3

3. Dilute

Dilute to 0.4 mg/ml with Actin Working Buffer (Cat. No. 000102). Add ATP to 0.2 mM and DTT to 0.5 mM.

Step 4

4. Incubate

Incubate the reconstituted actin on ice for 1 hour.

Step 5

5. Clarify

Clarify the reconstituted actin to remove any aggregates. Spin at 14,000 rpm for 15 minutes at 4°C in a microcentrifuge.

Step 6

6. Store

Add sodium azide to 0.05% and store at 4°C.

 NOTE :

The product is stable under these storage conditions for several weeks.