Can the Alkaline CometAssay® Control Cells be used for the Neutral CometAssay?

The Alkaline CometAssay Control Cells were qualified for the Alkaline CometAssay while the Neutral CometAssay Control Cells were qualified for the Neutral CometAssay.

Do control cells, Catalog # 4256-010-CC, just need to be put on a slide then stained? Or do they need to be ran through the entire Catalog # 4250-050-K protocol to see comets?

The control cells, Catalog # 4256-010-CC, need to be run through the entire protocol for Catalog # 4250-050-K in order to see comets.

How should the CometAssay® Control Cells be stored?

The CometAssay Control Cells should be stored in liquid nitrogen. To avoid the accumulation of damage due to freeze thaw cycles, the CometAssay Control Cells should be thawed and frozen in working aliquots. Refer to the CometAssay Control Cells literature for specific instructions.

What cell line is used to generate the CometAssay® Control Cells?

CometAssay Control Cells were prepared using an immortalized human lymphocyte line.

What is the purpose of the CometAssay® Control Cells?

The CometAssay Control Cells are designed to help standardize and compare alkaline and neutral electrophoresis methods between individual users and laboratories. The control cells contain known levels of damage.

Why are comet tails in positive control cells not present or smaller than expected?

Failure to lyse, denature in alkali, or properly perform electrophoresis may generate poor results. To improve results: Verify lysis of cells. Lysis Solution should be stored long term at room temperature and chilled to 4 °C prior to use. Lysis Solution stored long term at 4 °C will precipitate. Try overnight lysis at 4 °C. Perform alkali denaturation for 20 minutes at room temperature or for at least 1 hour at 4 °C. Prepare Alkaline Solution fresh from sodium hydroxide pellets. Electrophoresis conditions are also critical for optimal performance. Conventional slab gel electrophoresis chambers are not designed to eliminate known causes of comet assay variability (alkaline pH, buffer height, temperature, slide orientation). For optimal results, use a CometAssay Electrophoresis system (Catalog # 4250-050-ES). For insufficient electrophoresis time, increase time up to 1 hour at 4 °C for alkaline electrophoresis.

Why are there comet tails in the healthy CometAssay® Control Cell population?

The CometAssay Control Cells were prepared from an asynchronous cell population with cells at all stages of the cell cycle. Some percentage of the population will be undergoing DNA replication and may have strand breaks at replication forks. Improper storage may also lead to DNA damage. Do not store CometAssay Control Cells at -80°C. Store in aliquots in liquid nitrogen.

Why don't I see a visible cell pellet after centrifugation of the CometAssay® Control Cells?

A pellet may not be visible after centrifugation due to the low number of cells.

Why is it necessary to rinse the CometAssay® Control Cells in PBS before adding to LMAgarose?

The CometAssay Control Cells must be rinsed in PBS to eliminate media and storage factors that reduce adherence of LMAgarose to the slide. PBS without magnesium should be used, as magnesium stimulates nucleases.

CometAssay Alkaline Control Cells

Name: CometAssay Alkaline Control Cells
Brand: R&D Systems
SKU: 4256-010-CC

R&D Systems, part of Bio-Techne | Catalog # 4256-010-CC

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Product Summary
Preparation & Storage
Background
Product Documents

Key Product Details

Features

Cryopreserved cell preparations contain different levels of DNA damage to be used as controls when performing the comet assay.

Key Benefits

Provides a range of DNA damage for comparison with statistically distinct populations.
Allows for electrophoresis method to be standardized between individual users.
Standardized with the CometAssay Electrophoresis System
Cell populations remain stable in liquid nitrogen for long term storage.

Product Summary for CometAssay Alkaline Control Cells

What are CometAssay Alkaline Control Cells?

These cryopreserved cell preparations contain different levels of DNA damage to be used as controls when performing the comet assay. Cells are provided at 5 x 10^5 cells/mL. A healthy control cell population (C0) is treated with a DNA damaging agent to increase the amount of damage in populations C1, C2 and C3, respectively. Etoposide is the damaging agent used for Alkaline Control Cells (CC0-CC3).

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended on the product label.

CometAssay Alkaline Control Cells

R&D Systems, part of Bio-Techne | Catalog # 4256-010-CC

Key Product Details

Features

Key Benefits

Product Summary for CometAssay Alkaline Control Cells

Formulation, Preparation, and Storage

Shipping

Storage

Background: Apoptosis Detection

Alternate Names

Additional Apoptosis Detection Products

Product Documents for CometAssay Alkaline Control Cells

Product Datasheets

COA

Certificate of Analysis

SDS

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