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Index / SBI / pPACKH1-XL HIV Lentivector Packaging Kit /Product Detail : LV510A-1 pPACKH1-XL HIV Lentivector Packaging Kit
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#LV510A-1 pPACKH1-XL HIV Lentivector Packaging Kit

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  Price : 1286   EUR
1459   USD
997   GBP
5401   Zloty
172054   JPY
9916   NOK
10625   SEK
1454   CHF

Product name : pPACKH1-XL HIV Lentivector Packaging Kit

Catalog number : LV510A-1

Quantity: 50 transfections

Availability: Yes

Supplier name : SBI

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About this Product :

pPACKH1-XL HIV Lentivector Packaging Kit lentivectors random inserts of lentivectors are considered mot harmfull and the easiers way to get DNA into nulear DNA. More precise piggy back or talen techniques are used if random inserts are not wished. Our pPACKH1-XL HIV Lentivector Packaging Kit will randomly isert the DNA and since bad inserts are mostly lethal, healthy modofied cells can be studied.
http://antibody-antibodies.com/anti-elisa/Lentiviral.html

More Details about

Lenti Starter kit
Components:

1. pPACKH1-Plamid Packaging Mix (40μl)
2. PEG-it (5 ml)
3. TransDux (200μl at 200x)


What you will need:

1. Your Lentivector construct (3rd generation preferable)
2. HEK 293TN or FT cells and culture media
3. Tabletop low speed centrifuge (ex. Beckman GS-6R)

Protocol
Pseudovirus Production

Day 1
1. Plate 3x106 293TN cells in a fresh 10-cm plate in 10 ml of antibiotic free DMEM medium
(DMEM+FBS+Glu).

Day 2
2. The cells should be 50 to 70% confluent.
3. Transfect cells with plasmid containing gene of interest (2μg) and pPACKH1-plasmid mix
(10μg=20μl). Use Lipofectamine+Plus transfection reagent from Invitrogen (Follow
manufacturer’s protocol).


Day 4


4. Harvest viral supernate (culture media) and add PEG-it at a final volume of 1:5. Example: 2.5
ml of PEG-it should be added to 10 ml of viral supernate, invert 10 times to mix well. Keep
everything cold from this point onwards. Store viral Sup+ PEG-it at 40C overnight to 3 days.


Day 5


5. Harvest PEG-it precipitated virus by centrifuging at 40C at 1500xg for 30 min. Aspirate off the
supernate and resuspend the milky-white pellet in a small volume (30 to 50μl) of cold sterile
PBS or cold DMEM.
6. Freeze virus at -800C.

Transduction of Target Cells
TransDux protocol
Day 1


1. Plate 50,000 cells per well in a 24 well plate in culture medium.


Day 2


2. Cells should be between 50 to 70% confluent.
3. Aspirate medium from cells.
4. Add TransDux to complete medium to a final concentration of 1X (Example; add 60μl of
200x TransDux to 12 ml of medium – transfer 0.5ml of this mixture per 24 well)
5. Add virus to each well at different MOIs or different volumes.


Day 5


6. 72 hours post transduction, the viral genome will be integrated into the host cell genome.
7. Look at the cells for reporter expression if the viral construct has a reporter like GFP and/or
begin appropriate antibiotic selection to establish stable cell line.


OPTIONAL – Virus Titering


1. Aspirate off medium. Wash each well with PBS (at this point the plate can be frozen at -800C).
2. Add 100μl of Lysis Buffer (SBI’s Ultra Rapid Titer Kit) to each well.
3. Titer virus according to protocol given in the Ultra Rapid Titer Kit (SBI cat# LV960A-1).

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Pathways :
WP1004: Kit Receptor Signaling Pathway
WP1121: Kit Receptor Signaling Pathway
WP1341: Kit Receptor Signaling Pathway
WP147: Kit Receptor Signaling Pathway
WP407: Kit Receptor Signaling Pathway
WP548: neural crest development
WP774: Kit Receptor Signaling Pathway
WP886: Kit Receptor Signaling Pathway

Related Genes :
[KIT SCFR] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Piebald trait protein) (PBT) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (p145 c-kit) (v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog) (CD antigen CD117)
[Kit Sl] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit) (CD antigen CD117)
[KIT] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Proto-oncogene c-Kit) (Tyrosine-protein kinase Kit)
[KITLG MGF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG); Processed kit ligand]
[kit krk1] Mast/stem cell growth factor receptor-related protein Kit (EC 2.7.10.1) (Kit-related kinase 1) (xKrk1) (Tyrosine-protein kinase kit)
[KITLG MGF SCF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG)]
[Kitlg Kitl Mgf Sl Slf] Kit ligand (Hematopoietic growth factor KL) (Mast cell growth factor) (MGF) (Steel factor) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG)]
[Kitlg Kitl Mgf] Kit ligand (Hematopoietic growth factor KL) (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG)]
[KITLG SCF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Processed kit ligand]
[ygdG exo xni b2798 JW5446] Flap endonuclease Xni (FEN) (EC 3.1.-.-) (Exonuclease IX) (ExoIX)
[KITLG SCF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG)]
[ACSM1 BUCS1 LAE MACS1] Acyl-coenzyme A synthetase ACSM1, mitochondrial (EC 6.2.1.2) (Acyl-CoA synthetase medium-chain family member 1) (Butyrate--CoA ligase 1) (Butyryl-coenzyme A synthetase 1) (Lipoate-activating enzyme) (Middle-chain acyl-CoA synthetase 1) (XL-III) (Xenobiotic/medium-chain fatty acid:CoA ligase XL-3) (XM-ligase 3)
[kita kit sparse] Mast/stem cell growth factor receptor kita (SCFR) (EC 2.7.10.1) (Tyrosine-protein kinase Kit)
[gag-pol] Gag-Pol polyprotein (Pr160Gag-Pol) [Cleaved into: Matrix protein p17 (MA); Capsid protein p24 (CA); Spacer peptide 1 (SP1) (p2); Nucleocapsid protein p7 (NC); Transframe peptide (TF); p6-pol (p6*); Protease (EC 3.4.23.16) (PR) (Retropepsin); Reverse transcriptase/ribonuclease H (EC 2.7.7.49) (EC 2.7.7.7) (EC 3.1.26.13) (Exoribonuclease H) (EC 3.1.13.2) (p66 RT); p51 RT; p15; Integrase (IN) (EC 2.7.7.-) (EC 3.1.-.-)]
[hlyE clyA hpr sheA ycgD b1182 JW5181] Hemolysin E, chromosomal (Cytotoxin ClyA) (Hemolysis-inducing protein) (Latent pore-forming 34 kDa hemolysin) (Silent hemolysin SheA)
[IVa2] Packaging protein 1 (Packaging protein IVa2)
[KITLG MGF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG)]
[Kit KIT rCG_56893] Mast/stem cell growth factor receptor (EC 2.7.10.1)
[bcl-xL BCL2L1] Anti-apoptotic regulator Bcl-xL (Bcl-2-like protein 1)
[BAD BBC6 BCL2L8] Bcl2-associated agonist of cell death (BAD) (Bcl-2-binding component 6) (Bcl-2-like protein 8) (Bcl2-L-8) (Bcl-xL/Bcl-2-associated death promoter) (Bcl2 antagonist of cell death)
[KITLG MGF SCF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG); Processed kit ligand]
[KITLG SCF] Kit ligand (Mast cell growth factor) (MGF) (Stem cell factor) (SCF) (c-Kit ligand) [Cleaved into: Soluble KIT ligand (sKITLG); Processed kit ligand]
[gag-pol] Gag-Pol polyprotein (Pr160Gag-Pol) [Cleaved into: Matrix protein p17 (MA); Capsid protein p24 (CA); Spacer peptide 1 (SP1) (p2); Nucleocapsid protein p7 (NC); Transframe peptide (TF); p6-pol (p6*); Protease (EC 3.4.23.16) (PR) (Retropepsin); Reverse transcriptase/ribonuclease H (EC 2.7.7.49) (EC 2.7.7.7) (EC 3.1.26.13) (Exoribonuclease H) (EC 3.1.13.2) (p66 RT); p51 RT; p15; Integrase (IN) (EC 2.7.7.-) (EC 3.1.-.-)]
[kit] Mast/stem cell growth factor receptor Kit (SCFR) (EC 2.7.10.1) (Tyrosine-protein kinase kit)

Bibliography :
[22508377] Packaging HIV- or FIV-based lentivector expression constructs and transduction of VSV-G pseudotyped viral particles.