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.

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 Actin


Buffer


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

Unlabeled Tubulin


Pure Actin (Cat. No. 190101)

Recommended Products


Recommended Products


Unlabeled Tubulin


Pure Actin (Cat. No. 190101)

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 (21,000 RCF) 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.