ViraPower™ Zeo Lentivirus-Support-Kit

Das ViraPower™ Zeo Lentiviral Support Kit enthält die notwendigen Komponenten zur Transfektion von 293FT-Zellen, um virale Partikel zu erzeugen, dieWeitere Informationen

Katalognummer	Menge
K498500 auch als K4985-00 bezeichnet	20 Reaktionen

Katalognummer K498500

auch als K4985-00 bezeichnet

Preis (EUR)

2.060,00

Zum Warenkorb hinzufügen

Menge:

20 Reaktionen

Preis (EUR)

2.060,00

Zum Warenkorb hinzufügen

Das ViraPower™ Zeo Lentiviral Support Kit enthält die notwendigen Komponenten zur Transfektion von 293FT-Zellen, um virale Partikel zu erzeugen, die das Zielgen enthalten.Das Kit enthält den ViraPower™ Lentivirus-Verpackungsmix, Lipofectamine™ 2000 Reagenz und Zeocin™, und ist für die Verwendung mit einem ViraPower™ Lentivirus-Expressionsvektor konzipiert, der den Zeocin™-Marker zur stabilen Selektion enthält.Das Kit ist kompatibel mit allen unseren ViraPower™ lentiviralen Vektoren, einschließlich der ViraPower™ HiPerform™ lentiviralen Vektoren.

Kit enthält
• ViraPower™ lentivirale Verpackungsmischung
• Lipofectamine™ 2000 (Kat.-Nr. 11668027)
• Zeocin™-Lösung

Nur für Forschungszwecke.Nicht für therapeutische oder diagnostische Zwecke vorgesehen.

Nur für Forschungszwecke. Nicht zur Verwendung bei diagnostischen Verfahren.

Specifications

Konstitutives oder induktives SystemKonstitutiv

LiefertypLentiviral

Zur Verwendung mit (Anwendung)Virale Expression

ProdukttypLentivirus-Support-Kit

Menge20 Reaktionen

Selektionsmittel (eukaryotisch)Blasticidin

VektorpLP

KlonierungsmethodeGateway™

ProduktlinieViraPower

PromoterRSV, CMV

ProteinmarkierungV5-Epitop-Tag

Unit SizeEach

Inhalt und Lagerung

Inhalt
Das ViraPower™ Zeo Lentivirus-Support-Kit enthält die folgenden Komponenten:
• 195 µg ViraPower™ Verpackungsmix (enthält 195 µl einer Mischung aus pLP1-, pLP2- und pLP⁄VSVG-Plasmiden bei 1 µg⁄µl in TE-Puffer, pH 8,0)
• 0,75 ml Lipofectamine™ 2000 (Kat.-Nr. 11668027)
• 125 mg Zeocin™ (1,25 ml einer 100 m⁄ml-Lösung)

Lagerung
• ViraPower™ Verpackungsmix: -20 °C
• Lipofectamine™ 2000: 4 °C (nicht einfrieren)
• Zeocin™: -20 °C, lichtgeschützt

Häufig gestellte Fragen (FAQ)

I used one of your lentiviral vectors but am observing cytotoxic effects after transduction. Can you please help?

Possible causes include:

- large volume of viral supernatant used for transduction
- cells sensitive to Polybrene regaent
- too much antibiotic used for selection
- antibiotic used too soon after tranduction
- gene of interest is toxic to cells

I transduced my lentiviral stock into my mammalian cell line but am getting poor expression of my gene of interest. What could have happened?

Poor expression could result from low transduction efficiency, too low of a MOI, too much antibiotic used for selection, usage of antibiotic too soon after transduction, harveting cells too soon after transduction, having a gene of interest that is toxic to cells, or rerrangement in the LTR regions of the expression construct plasmid DNA.

I transduced my lentiviral stock into my mammalian cell line but am getting no expression of my gene of interest. What could have gone wrong?

Here are some possible causes and solutions:
- Promoter silencing; CMV promoter is prone to silencing especially in mouse and rat cells, screen multiple antibiotic resistant clones and select the one with the highest expression levels
- Viral stocks stored incorrectly; aliquot and store at -80 degrees C, do not freeze/thaw more than 3 times

I prepared a lentiviral stock using one of your lentiviral vectors. I am trying to determine the titer using antibiotic selection but am not able to since the cells are very confluent and I am not getting antibiotic-resistant clones. Can you please offer some tips?

Here are some possible causes and solutions:

- Too little antibotic used for selection
- Selection performed on confluent cells; replate cells
- Viral supernatant not diluted sufficiently; titer lentivus using a wider range of 10-fold serial dilutions

I am using one of your lentiviral vectors and am getting a low lentiviral titer. Can you offer some troubleshooting tips?

Possible causes include:

- low transfection efficiency; Use a high-quality plasmid prep, 293FT cells under passage 16, ensure removal of Geneticin during transfection, ensure correct DNA:lipid ratio, and that cells are plated at the correct confluency
- transfected cells are not cultured in medium containing sodium pyruvate; this reagent provides an extra energy source for cells
- viral supernatant harvested too early; viral supernatants can generally be collected 48-72 hrs post-transfection
- viral supernatant too dilute; concentrate virus using CsCl purification
- viral supernatant frozen and thawed multiple times; 3 times should be the maximum freeze/thaw
- gene of interest is large; viral titers decrease as size of insert increases, inserts larger than 5.6 kb are not recommended
- rearrangement in the LTR region of the epxression construct plasmid DNA; use Stb3 cells for transformatin of the lentiviral construct
- poor choice of titering cell line; use HT1080 cells or similar cell line
- Polybrene reagent is not included during transduction; transduce lentiviral construct into cells in the presence of Polybrene reagent
- Lipofectamine reagent handled incorrectly; ensure proper storage and mix gently before use
- Use fluorescence micrscopy to check titer with HiPerform FastTiter lentivirus

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

View all ViraPower™ Zeo Lentivirus-Support-Kit FAQs