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.
Fortfahren mit
CAPHOS
Calciumphosphat-Transfektionskit
Most cost effective transfection reagent kit for transient and stable transfection of DNA into mammalian cells
Synonym(e):
Gene delivery
Anmelden zur Ansicht der Organisations- und Vertragspreise.
Größe auswählen
Ansicht ändern
| Ihnen/SKU | Verfügbarkeit | Preis |
|---|---|---|
1 kit | Warenkorb auf Verfügbarkeit prüfen | CHF 582.00 |
Über diesen Artikel
UNSPSC Code:
12352200
EC Number:
233-140-8
NACRES:
NA.85
Form:
solution
Grade:
Molecular Biology
Technique(s):
transfection: suitable
CHF 582.00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
Benötigen Sie Hilfe? Unser Team von erfahrenen Wissenschaftlern ist für Sie da.
Unterstützung erhaltengrade
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
Die Calciumphosphat-Transfektion ist eine häufig genutzt Methode für das Einführen von DNA in eukaryotische Zellen. Diese Methode wird verwendet, und in einer großen Vielfalt von Zellarten sowohl transiente als auch stabile Transfektionen zu erzielen.
Application
Das Calciumphosphat-Transfektionskit hat folgende Anwendungen:
- für Transfektionsverfahren
- für die Transfektion von Hek293T-Zellen
- für die Transfektion von H29D-Zellen
Geeignet für transiente wie für stabile Transfektionen von DNA in kultivierte Säugetierzellen. Bei den folgenden Zellen wurde eine erfolgreiche Transfektion mit der Calciumphosphat-Methode durchgeführt:
BAEC
Melanomzellen des Darms
CHO K1
COS-7
Fibroblasten (human embryonal, Neo Derm)
HEK293
Huh 7
IMR-90
LLC (Lewis-Lungenkarzinom)
NIH3T3
PC-12
PCI-13
SH-Sy5Y
SK-Hep-1
T47D
BAEC
Melanomzellen des Darms
CHO K1
COS-7
Fibroblasten (human embryonal, Neo Derm)
HEK293
Huh 7
IMR-90
LLC (Lewis-Lungenkarzinom)
NIH3T3
PC-12
PCI-13
SH-Sy5Y
SK-Hep-1
T47D
Biochem/physiol Actions
Bei diesem Verfahren wird eine HEPES-gepufferte Kochsalzlösung, die Natriumphosphat enthält, langsam mit einer CaCl2-Lösung gemischt, in der sich die DNA befindet. Es bildet sich ein DNA-Calciumphosphat-Co-Präzipitat, das an der Zelloberfläche anhaftet und von der Zelle aufgenommen wird, mutmaßlich mittels Endozytose.
Features and Benefits
- Geeignet für transiente wie für stabile Transfektionen
- Reproduzierbar für eine breite Palette von Zellarten
- Häufig referenziert
- Kostengünstig
Other Notes
Das Protokoll wurde entwickelt für die transiente Transfektion von CHO-Zellen unter Verwendung eines pSV40-CAT-Plasmids als Reportergen.
Lieferumfang des Calciumphosphat-Transfektionskits:
5 ml 2,5 M CaCl2 (C2052)
25 ml 2x HEPES-gepufferte Kochsalzlösung (H1012)
25 ml Wasser in Molekularbiologie-Qualität (W4502)
5 ml 2,5 M CaCl2 (C2052)
25 ml 2x HEPES-gepufferte Kochsalzlösung (H1012)
25 ml Wasser in Molekularbiologie-Qualität (W4502)
1 of 1
Dieser Artikel | |||
|---|---|---|---|
| grade Molecular Biology | grade Molecular Biology | grade - | grade - |
| technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) microbe id | staining: suitable | technique(s) transfection: suitable |
| form solution | form dried film | form - | form powder |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level - |
| storage temp. −20°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −20°C |
| shipped in dry ice | shipped in - | shipped in dry ice | shipped in dry ice |
signalword
Warning
hcodes
Hazard Classifications
Eye Irrit. 2
Lagerklasse
12 - Non Combustible Liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
wgk
WGK 1
Hier finden Sie alle aktuellen Versionen:
Besitzen Sie dieses Produkt bereits?
In der Dokumentenbibliothek finden Sie die Dokumentation zu den Produkten, die Sie kürzlich erworben haben.
-
Can antibiotics be present in the medium during transfection?
1 answer-
Helpful?
-
-
What is the Department of Transportation shipping information for this product?
1 answer-
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.
Helpful?
-
-
Is the size of the plasmid an important consideration for transfection?
1 answer-
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.
Helpful?
-
-
Is optimizing the transfection protocol important?
1 answer-
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.
Helpful?
-
-
Is low cell passage number an important consideration for transfection?
1 answer-
Yes, we recommend cells are at a low passage when being used for any application, including transfection. The reason why depends on what type of cells they are. Primary cells will undergo a finite number of divisions, and as they get closer to senesence they divide more slowly - both affecting their ability to take up DNA (transient transfection), and minimizing their abillity to incorporate the DNA into the genome (stable selection).Cultured common cell lines are often immortalized, and generally continue to aquire mutations, leading to a heterogenous population that may perform differently from cells of lower passage number - leading to results that are not reproducible.
Helpful?
-
-
How can I increase the efficiency of the calcium phosphate transfection method?
1 answer-
The calcium phosphate transfection method is performed differently than other methods, and so it can be optimized beyond the general recommendations. An important part of the protocol is generation of a fine precipitate. Be sure that the bubbling in tube B is consistant and with good force, while the solution from tube A is added very slowly, dropwise. Once the precipitates are added to the cell culture, be sure they are evenly distributed on the plate so all cells may be transfected.The pH of the HeBS is critical to precipitate formation. Prolonged storage of the solution may cause the pH to shift away from the optimal 7.05 - 7.12.
Helpful?
-
-
What is transfection efficiency?
1 answer-
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.
Helpful?
-
-
Can I transfect cells plated at low density?
1 answer-
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.
Helpful?
-
-
Why do I see a precipitate in my cell culture after calcium-phosphate transfection?
1 answer-
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.
Helpful?
-
-
How do I choose a transfection reagent?
1 answer-
There are many guides that help you select a transfection reagent. In general, consider:The type of cell(s) you will transfectThe type of nucleic acid or protein you will introduce to the cellThe composition of your cell culture mediumThe need for stable or transient transfectionThe equipment you have availableThe other factors important to you - cost, protocol flexibility, ease of use, etc.
Helpful?
-