Calculating transfection efficiency is very useful when optimizing transfection protocols. Transfection efficiency can be performed using a GFP-expressing plasmid. After transfection, cells are stained with propidium iodide and counted. The propidium iodide provides a count of the total cells in the population, and the GFP-expressing cells provide a count of the number of cells transfected. The transfection efficiency (%) can then be calculated by:(# GFP-expressing cells / total cell #) * 100
CAPHOS
リン酸カルシウムトランスフェクションキット
Most cost effective transfection reagent kit for transient and stable transfection of DNA into mammalian cells
別名:
Gene delivery
ログインで組織・契約価格をご覧ください。
サイズを選択してください
表示を変更する
| あなたへ/カタログ番号 | 在庫状況 | 単価 |
|---|---|---|
1 kit | カートの在庫状況を確認する | ￥107,000 |
この商品について
UNSPSC Code:
12352200
EC Number:
233-140-8
NACRES:
NA.85
Form:
solution
Grade:
Molecular Biology
Technique(s):
transfection: suitable
￥107,000
カートの在庫状況を確認する
grade
Molecular Biology
Quality Segment
form
solution
usage
kit sufficient for 160 transfections (6 cm dishes), kit sufficient for 400 transfections (3.5 cm dishes), kit sufficient for 80 transfections (10 cm dishes)
technique(s)
transfection: suitable
shipped in
dry ice
storage temp.
−20°C
General description
リン酸カルシウムトランスフェクションは真核生物細胞へのDNA導入に通常使用される方法です。この技術は多種多様な細胞タイプで、一時的かつ安定なトランスフェクションを得るために使用されています。
Application
リン酸カルシウムトランスフェクションキットの用途：
- トランスフェクションの可能化
- Hek293T細胞のトランスフェクション
- H29D細胞のトランスフェクション
DNAの培養哺乳動物細胞中への一時的で安定なトランスフェクションに適しています。以下の細胞はリン酸カルシウム法を用いて成功裏に形質移入されています：
BAEC
腸黒色腫細胞
CHO K1
COS-7
線維芽細胞（ヒト胚性、新真皮）
HEK293
Huh 7
IMR-90
LLC（ルイス肺癌）
NIH3T3
PC-12
PCI-13
SH-Sy5Y
SK-Hep-1
T47D
BAEC
腸黒色腫細胞
CHO K1
COS-7
線維芽細胞（ヒト胚性、新真皮）
HEK293
Huh 7
IMR-90
LLC（ルイス肺癌）
NIH3T3
PC-12
PCI-13
SH-Sy5Y
SK-Hep-1
T47D
Biochem/physiol Actions
手順は、リン酸ナトリウムを含むHEPES緩衝生理食塩水を、DNAを含むCaCl2溶液とゆっくり混合することに基づいています。おそらくエンドサイトーシスによって細胞表面に接着し細胞に取り入れられるDNA-リン酸カルシウム共沈物が生じます。
Features and Benefits
- 一時的で安定なトランスフェクションに適しています
- 広範囲の細胞タイプに関して再現性があります
- 広く参照されています
- 安価です
Other Notes
プロトコルは、pSV40-CATプラスミドをレポーター遺伝子として用いるCHO細胞の一時的なトランスフェクションのために策定されました。
リン酸カルシウムトランスフェクションキットには以下のものが含まれます：
5 mL 2.5M CaCl2 （C2052）
25 mLのHEPES緩衝生理食塩水が2つ（H1012）
25 mLの分子生物学グレード水（W4502）
5 mL 2.5M CaCl2 （C2052）
25 mLのHEPES緩衝生理食塩水が2つ（H1012）
25 mLの分子生物学グレード水（W4502）
1 of 1
当該品目 | |||
|---|---|---|---|
| description Most cost effective transfection reagent kit for transient and stable transfection of DNA into mammalian cells | description - | description Avanti Research™ - A Croda Brand | description Reagent for transient & stable DNA transfections |
| form solution | form - | form powder | form powder |
| grade Molecular Biology | grade - | grade - | grade Molecular Biology |
| technique(s) transfection: suitable | technique(s) microbe id | staining: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable |
| Quality Level 200 | Quality Level 200 | Quality Level - | Quality Level 100 |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. room temp |
| usage kit sufficient for 400 transfections (3.5 cm dishes), kit sufficient for 80 transfections (10 cm dishes), kit sufficient for 160 transfections (6 cm dishes) | usage kit sufficient for 100 tests (using a 3.5 cm dish) | usage - | usage - |
signalword
Warning
hcodes
Hazard Classifications
Eye Irrit. 2
保管分類
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
wgk
WGK 1
適用法令
試験研究用途を考慮した関連法令を主に挙げております。化学物質以外については、一部の情報のみ提供しています。 製品を安全かつ合法的に使用することは、使用者の義務です。最新情報により修正される場合があります。WEBの反映には時間を要することがあるため、適宜SDSをご参照ください。
キットコンポーネントの情報を参照してください
pdsc
キットコンポーネントの情報を参照してください
prtr
キットコンポーネントの情報を参照してください
fsl
キットコンポーネントの情報を参照してください
ishl_indicated
キットコンポーネントの情報を参照してください
ishl_notified
キットコンポーネントの情報を参照してください
cart
キットコンポーネントの情報を参照してください
jan
-
How can I determine the efficiency of my transfection?
1 回答-
役に立ちましたか？
-
-
Is optimizing the transfection protocol important?
1 回答-
For many common cell lines, transfection reagent efficiency is very high and the protocols will not require any optimization. For hard-to-transfect cells or those ultimately expressing a toxic protein, the protocol should be optimized for best transfection efficiency. Taking time to optimize will give you more transfected cells with each procedure, which can mean more protein expressed and results that are reproducible.
役に立ちましたか？
-
-
What is transfection efficiency?
1 回答-
Transfection efficiency is a measure of how many cells take up the DNA during the transfection process. Many transfection reagents can achieve a transfection efficiency of >90% in common cell lines. Other cell lines are hard to transfect, and require special reagents and/or techniques to achieve even a small population of transfected cells.
役に立ちましたか？
-
-
What is the difference between stable and transient transfection?
1 回答-
When the DNA enters the nucleus of the cell, the plasmid is replicated by the cell machinery (transient transfection). During this time, RNA is transcribed and protein translated until the plasmid DNA is lost after a few cell divisions. This expression of the plasmid DNA, mRNA, and protein is transient (temporary).In some cases, the plasmid DNA is integrated into the host cell genome. This is usually accompanied by forced expression using a selection antibiotic and sometimes a cloning step (to be sure all cells have the same integration site). Once the DNA is stable, the cell line can be frozen and used to express protein for many years. Clones may even be screened for those expressing the highest amount of protein.
役に立ちましたか？
-
-
How can I increase the efficiency of my transfection?
1 回答-
Transfection efficiency is affected by many different things, including plasmid size and purity, media components present, transfection reagent selected, amount of DNA and transfection reagent used, cell density, etc. Optimizing the protocol with respect to these concerns will allow you to achieve a higher transfection efficiency. For many cell lines and transfection reagents, optimized protocols are already available.
役に立ちましたか？
-
-
What is the Department of Transportation shipping information for this product?
1 回答-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
役に立ちましたか？
-
-
Is the size of the plasmid an important consideration for transfection?
1 回答-
The size of the plasmid should be considered when selecting a transfection reagent with the best efficiency. In general, larger sized plasmids should easily transfect with readily available transfection reagents, as along as the plasmid DNA is of high purity.
役に立ちましたか？
-
-
Can I transfect cells plated at low density?
1 回答-
For most transfections, cells should be >70% confluency the day of transfection, and growing in mid-log phase. Some transfection reagents are now designed to work with cells at low density, when required.
役に立ちましたか？
-
-
Can antibiotics be present in the medium during transfection?
1 回答-
We recommend that no antibiotics are present during transfection. The process of transfection can make the cells somewhat more porous to allow for efficient DNA entry. During this time, antibiotics will also enter the cells more easily and the cells may show increased cell death. Wait until about 24 hours after transfection to resume the use of preventative antibiotics and/or start the use of selective antibiotics.
役に立ちましたか？
-
-
Why do I see a precipitate in my cell culture after calcium-phosphate transfection?
1 回答-
The precipitate is normal to see - this is the calcium/DNA precipitate the cell will internalize during the transfection process. If bubbled correctly, the precipitates will be very fine and regular sized, then evenly distributed over the cells.
役に立ちましたか？
-