ACT-1

The ACT-1 cell line originates from a strain of the slime mold Dictyostelium discoideum, which serves as a valuable model for studying actin dynamics and cellular motility. In Dictyostelium, actin plays a pivotal role in cytoskeletal organization, particularly in the process of chemotaxis and cellular aggregation under nutrient deprivation. The ACT-1 cell line has been essential for analyzing monoclonal antibody interactions with actin, especially in distinguishing specific binding regions on actin monomers and assessing actin's structural and functional responses to various molecular interactions. Studies using ACT-1 have contributed significantly to understanding how actin's amino-terminal regions interact with antibodies and influence actin polymerization, highlighting regions crucial for monomer-to-filament transitions and stabilization.

In particular, ACT-1 has been used to examine monoclonal antibodies that can prevent actin polymerization by binding selectively to monomeric forms of actin. This binding inhibits filament formation by stabilizing actin in its monomeric state, which reduces filament length and viscosity when introduced to existing actin filaments. These effects are useful for studying actin-regulating proteins and their mechanisms in cellular processes. Given its specificity, the ACT-1 line is instrumental for both biochemical assays and immunofluorescence studies, which utilize anti-actin monoclonal antibodies to map structural actin domains and assess actin-binding proteins’ regulatory roles in cellular architecture.

37°C, 5% CO₂, humidified atmosphere.

1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.

To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.

For full details, please refer to our Warranty Statement.