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ClonaCell™tcs /潮霉素工具包

试剂盒包含ClonaCell™-TCS培养基和潮霉素B抗生素
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产品号 #（选择产品）

产品号 #03823_C

试剂盒包含ClonaCell™-TCS培养基和潮霉素B抗生素

产品优势

  • 支持多种细胞系的高克隆效率和强大的集落形成，如：CHO-K1， CHO-S, HEK-293, B16F-10, BaF/3, BHK-1, FD-5, Jurkat Daudi, Molt-4, UT-7, K562

产品组分包括

  • ClonaCell™-TCS培养基，80 mL（目录#03814）
  • 湿霉素B， 100毫克（目录号03813）
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概述

本试剂盒用于湿霉素B抗生素作为选择剂的选择和克隆应用。ClonaCell™-TCS Medium是一种含血清、以甲基纤维素为基础的半固体培养基，可用于选择和克隆各种悬浮适应和贴壁细胞系，如BHK-1、CHO-K1、CHO-S、CHO-DG44和HEK-293。该培养基含有预先选择的胎牛血清（FBS）和牛血清白蛋白（BSA）。当使用ClonaCell™-TCS培养基时，潮霉素B适用于细胞培养中对潮霉素耐药克隆的选择和维持。

Subtype
Specialized Media
 
Cell Type
CHO Cells, HEK-293, Other
 
Species
Mouse, Other
 
Application
Cell Culture, Semi-Solid Cloning
 
Brand
ClonaCell
 
Area of Interest
Antibody Development, Cell Line Development
 

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Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet 1
Product Name
ClonaCell™-TCS/Hygromycin Kit
Catalog #
03823
Lot #
All
Language
English
Document Type
Product Information Sheet 2
Product Name
ClonaCell™-TCS/Hygromycin Kit
Catalog #
03823
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
ClonaCell™-TCS/Hygromycin Kit
Catalog #
03823
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
ClonaCell™-TCS/Hygromycin Kit
Catalog #
03823
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (2)

ClonaCell™ Media for Hybridoma and Cell Line Development
Brochure
ClonaCell™ Media for Hybridoma and Cell Line Development
Protocol for Cloning CHO Cell Lines Using ClonaCell™-CHO Semi-Solid Medium
Technical Bulletin
Protocol for Cloning CHO Cell Lines Using ClonaCell™-CHO Semi-Solid Medium

Frequently Asked Questions

Why do I get more cells when I select my fusion in liquid medium rather than in methylcellulose-based semi-solid medium?

Cells grown in liquid medium may appear to grow more rapidly than in methylcellulose-based medium. This is often due to the presence of a few rapidly growing clones that multiply quickly and become abundant in liquid culture, overgrowing clones that grow more slowly. In methylcellulose cultures, the rapidly growing cells remain in close proximity to each other, resulting in large colonies, each derived from a single fusion or transfection product. The large clones don't overgrow smaller, slower growing colonies, which can be separately isolated.

How do I thaw ClonaCell™ methylcellulose-based semi-solid medium?

We recommend thawing the medium overnight in a refrigerator at 4°C and mixing well.

How do I measure and dispense methylcellulose semi-solid medium?

We recommend using a 12 mL syringe with a 16 gauge needle attached (blunt end needles are recommended for safety purposes). Do not dispense the semi-solid media/cell mixture using serological pipettes as the media will stick to the pipette walls, resulting in inaccurate dispensed volumes and loss of cells.

My ClonaCell™ methylcellulose semi-solid medium appears runny. Why does this happen?

"Runny" methylcellulose could be a result of improper handling. Diluting the methylcellulose with too much liquid medium, or insufficient mixing before use, will result in methylcellulose with altered viscosity. Excessive condensation on the inside of the cell culture dish lid can result in water dripping onto the cultures, lowering viscosity. Additionally, bumping, shaking or other sudden movement of the culture may also disrupt the colonies. Note: methylcellulose is less viscous at room temperature than at 37°C.

What is the optimal number of colonies per plate?

We recommend 50-150 colonies per plate. As it is difficult to anticipate the numbers of colonies after a fusion or transfection, we recommend plating at three different densities to increase the likelihood of achieving a plating density of approximately 100 colonies per plate. This density allows sufficient space between the colonies to allow for easy colony picking.

There are still bubbles in the media after I plate my cells. Do I need to disrupt the bubbles?

We recommend that you avoid creating large bubbles during plating, but there is no need to manually pop or disperse the small bubbles after plating. They will disperse over the incubation period of 10-14 days.

Do I ever need to re-clone cultures grown with ClonaCell™ semi-solid medium?

Re-cloning is a good practice to observe and is recommended if the number of colonies in the original dishes was very high.

Once I pick the colonies and grow the cells in plates, will the residual methylcellulose interfere with characterization? For example, will I have problems doing an ELISA?

There will likely be some residual methylcellulose contamination when colonies are picked and transferred to the 96-well plate with the liquid growth medium. The concentration of methylcellulose, however, should be low enough that it should not interfere with most assays.

How important is the incubator humidity when culturing in methylcellulose-based medium?

Very important. In situations where the humidity is not high enough, we recommend that the 100 mm Petri dishes should be placed with an open dish containing sterile water inside a larger plastic container with a lid. Without very high humidity, the media will dry out over the culture period and this will impede the growth of the colonies.

Do I have to use 100 mm petri dishes or can I use other cultureware?

We recommend 100 mm Petri dishes as these have been used to develop and test ClonaCell™ semi-solid media. We have found that the surface area of these dishes allows for easy colony picking. Other sizes of dish (e.g. 6-well plates) can be used. It is important to use non-coated dishes to prevent cells from sticking to the bottom of the plate and obscuring the colonies. The volume of media plated should be adjusted to reflect the surface area of the dish being used.

Is the serum in ClonaCell™-TCS medium heat inactivated?

Yes, all serum used in ClonaCell™ is heat inactivated.

Is there any IgG in ClonaCell™ TCS?

While we don't add IgG to the ClonaCell™ media, we do add serum, which contains an undefined amount of IgG. We selectively use serum lots with low IgG levels in the production of ClonaCell™ media, however, levels vary from lot to lot. IgG levels in a specific lot of ClonaCell™ TCS medium are available in the lot-specific Certificate of Analysis.

Can ClonaCell™-TCS be used with any cell line?

A list of recommended cell lines can be found in the manual. Other cell lines may be compatible with ClonaCell™-TCS. It will be necessary, however, to determine the plating cell density and growth efficiency of the desired cells in ClonaCell™-TCS.

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Species Mouse, Other
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