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[Protocol] Cell-Type Specific Transfection Protocols (X-treme)

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작성자 다인바이오 작성일14-10-10 10:22 조회1,654회 댓글0건

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Cell-Type Specific Transfection Protocols



Experiments below show high level gene expression in mammalian and insect cells transfected using X-tremeGENE Transfection Reagents.

Fast straightforward transfection protocols can be obtained by clicking the cell type below. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.

Protocols and results provided by customers
Protocols shown here are provided by customers and have not been verified by Roche scientists. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.





X-tremeGENE HP DNA Transfection Reagent


Cell name
(Click to view protocol)

Cell type

Transfected reporter gene

Customer protocol(s)

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

GFP / EGFP-CB2 / Not indicated

available

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

Stable transfection

available

293-A

Human embryonic kidney cell line

GFP

-

293-F

Human embryonic kidney cell line subclone

Luciferase

-

293FT

Human embryonic kidney cell line subclone

GFP

available

293T

Human embryonic kidney cell line expressing SV40 large T antigen

GFP / Interleukin / Fc fusion protein

available

3T3 (ATCC®CRL-1658™)

Mouse embryonic fibroblasts

GFP

available

3T3-L1 (ATCC®CRL-173™)

Mouse embryonic fibroblasts

SEAP

-

A549 (ATCC®CCL-185™)

Human carcinomic alveolar basal epithelial cells

SEAP and EGFP

available

AGS (ATCC®CRL-1739™)

Human epithelial adenocarcinoma cells

GFP

available

C2C12 (ATCC® CRL-1772™)

Mouse myoblast cell line

SEAP and EGFP

available

CHO/dhFr- (ATCC® CRL-9096™)

Chinese hamster ovary cell line subclone

Secretable protein

available

CHO-K1 (ATCC® CCL-61™)

Chinese hamster ovary cell line subclone

GFP

available

CHO-S

Chinese hamster ovary cell line subclone

Luciferase

-

COS-7 (ATCC® CRL-1651™)

African green monkey kidney fibroblast-like cell line

GFP

available

Dental papilla mesenchymal cells

Primary cells from the first lower molar of E18.5 Swiss mice

GFP

available

EC109

Esophageal squamous carcinoma cell line

Not indicated

available

EC9706

Esophageal squamous carcinoma cell line

Not indicated

available

F9 (ATCC® CRL-1720™)

Mouse embryonal carcinoma cells

EGFP / GFP

available

Fibroblasts, primary

Human primary fibroblasts

GFP

-

H1

Human embryonic stem cells

GFP

-

HCT 116 (ATCC® CCL-™)

Human colorectal carcinoma cell line

Luciferase

-

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

GFP

-

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP

available

HepG2.2.15

Human hepatoma cell line expressing complete HBV genome

GFP

available

High Five

Insect cell line (Invitrogen)

ß-gal

-

HT-29 (ATCC® HTB-38™)

Human colorectal adenocarcinoma cell line

GFP

-

HT-1080 (ATCC® CCL 121™)

Human fibrosarcoma cell line

GFP

available

HuH-7 (ATCC® JCRB-0403)

Human hepatoma cell line

GFP

-

HuH-7.5

Human hepatoma cell line

EGFP

available

Jurkat

Human lymphoblast cell line

GFP

-

K-562 (ATCC® CCL 243™)

Human immortalised myelogenous leukemia cell line

GFP

-

L02

Immortal hepatic cell line

RNAi plasmid encoding GFP

available

LNCaP (ATCC® CRL-1740™)

Human prostate cancer cell line

GFP

available

MC3T3-E1 (ATCC® CRL-2593™)

Mouse osteoblast-like cells

sheep C12orf29 protein

-

MCF7 (ATCC® HTB-22™)

Human adenocarcinoma cell line

GFP

-

MDA-MB-231 (ATCC® HTB-26™)

Human adenocarcinoma cell line

SEAP

-

MEF

Primary mouse embryonic fibroblasts

GFP

-

MRC-5

Human lung fibroblast cells

GFP

-

PC-3 (ATCC® CRL-1435™)

Human prostate adenocarcinoma cell line

GFP

-

PC-12 (ATCC® CRL-1721™)

Rat adrenal pheochromocytoma cell line

Luciferase

-

Plat-E

Human kidney packaging cell line

GFP

available

Porcine ovarian granulosa cells

Primary cells

EGFP

available

PK-15 (ATCC®CCL-33™)

Porcine kidney epithelial cell line

GFP

available

RAW 264.7 (ATCC® TIB-71™)

Mouse monocyte macrophage cell line

GFP

-

Schneider S2-R+

Epithelial Drosophila melanogaster cell line

GFP

-

Sf9 (ATCC® TIB-71™)

Epithelial fall armyworm (Spodoptera frugiperda) cells

ß-gal

-

Stems cells, mesenchymal

Human mesenchymal stem cells

GFP

-

Stems cells, mesenchymal

Murine mesenchymal stem cells (RM26/2)

Far-red fluorescent protein

available

Stems cells, mesenchymal

Murine mesenchymal stem cells (EK8)

Far-red fluorescent protein

available

Stems cells, mesenchymal

Murine mesenchymal stem cells (p53-/-)

Far-red fluorescent protein

available

U-2 OS (ATCC® HTB-96™)

Human osteosarcoma cell line

GFP

available

U-87 MG (ATCC® HTB-14™)

Human epithelial glioblastoma cells

GFP

available


For further information on cells transfected with X-tremeGENE HP DNA Transfection Reagent you can also browse the Transfection Database.


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X-tremeGENE 9 DNA Transfection Reagent


Cell name
(Click to view protocol)

Cell type

Transfected reporter gene

Customer protocol(s)

293-A

Human embryonic kidney cell line

EGFP

available

293FT

Human embryonic kidney cell line subclone

Luciferase

available

293T

Human embryonic kidney cell line

Luciferase / EGFP / Flag-tagged protein

available

BF-2 (ATCC® CCL-91™)

Human embryonic kidney cell line

RFP and EGFP

available

C2C12 (ATCC® CRL-1772™)

Mouse myoblast cell line

EGFP

available

CHO-K1 (ATCC® CCL-61™)

Chinese hamster ovary cell line subclone

GFP

-

CHSE-214 (ATCC® CRL-1681™)

Chinook salmon embryo (fish cell line)

EGFP

available

COS-7 (ATCC® CRL-1651™)

African green monkey kidney fibroblast-like cell line

GFP

available

EPC (ATCC® CRL-2872™)

Fish cell line (Epithelioma papulosum cyprini)

RFP

available

ES-E14TG2a (ATCC® CRL-8121™)

Mouse embyonic stem cells

GFP

-

GF

Fish cell line (grouper fin)

RFP and EGFP

available

HCT 116 (ATCC® CCL-™)

Human colorectal carcinoma cell line

Luciferase

-

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

EGFP-actin

available

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

GFP

-

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP

-

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

14 kb plasmid (incl. eGFP; stable)

available

HINAE

Japanese flounder (Hirame Natural Embryo) fish cell line

RFP and EGFP

available

HT-1080 (ATCC® CCL-121™)

Human fibrosarcoma cell line

GFP

-

HuH-7 (ATCC® JCRB-0403)

Human hepatoma cell line

Luciferase

available

MCF7 (ATCC® HTB-22™)

Human adenocarcinoma cell line

GFP

-

Neuro-2a (ATCC® CCL-131™)

Mouse neuroblastoma cells

GFP-mortalin fusion protein

available

NIH-3T3 (ATCC® CRL-1658™)

Mouse embryonic cell line (fibroblast)

GFP

-

PC-3 (ATCC® CRL-1435™)

Human prostate adenocarcinoma cell line

GFP

-

PC-12 (ATCC® CRL-1721™)

Rat adrenal pheochromocytoma cell line

Luciferase

-

Primary pulmonary fibroblasts

Human pulmonary fibroblasts (primary)

Not indicated

available

Porcine granulosa cells

Porcine granulosa cells (subculture of primary cells)

GFP fusion protein

available

Sf9 (ATCC® TIB-71™)

Epithelial fall armyworm (Spodoptera frugiperda) cells

GFP

available


For further information on cells transfected with X-tremeGENE 9 DNA Transfection Reagent you can also browse the Transfection Database.


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X-tremeGENE siRNA Transfection Reagent


Cell Type

X-tremeGENE
siRNA Reagent

siRNA1

Culture Dish

Total Volume

CHO-K1 (ATCC® CCL-61™)

10 µl

2 µg

24-well plate

550 µl

COS-7 (ATCC® CRL-1651™)

2.5 µl

0.5 µg

24-well plate

550 µl

HEK-293 (ATCC® CRL-1573™)

5 µl

0.5 µg

24-well plate

550 µl

HeLa (ATCC® CCL-2™)

5 µl

0.5 µg

24-well plate

550 µl

Hep G2 (ATCC® HB-8065™)

10 µl

2 µg

24-well plate

550 µl

HT-29 (ATCC® HTB-38™)

10 µl

2 µg

24-well plate

550 µl

HTC-116 (ATCC® CCL-247™)

2.5 µl

0.5 µg

24-well plate

550 µl

Huh-7 clone 9B

10 µl

1 µg

6-well plate

1000 µl2

MCF7 (ATCC® HTB-22™)

2.5 µl

0.5 µg

24-well plate

550 µl

NIH 3T3 (ATCC® CRL-1658™)

5 µl

0.5 µg

24-well plate

550 µl

PC-3 (ATCC® CRL-1435™)

5 µl

0.5 µg

24-well plate

550 µl

PC-12 (ATCC® CRL-1721™)

2.5 µl

0.5 µg

24-well plate

550 µl


10.5 µg corresponds to 67.5 nM siRNA
2medium changed 5 hours after transfection

For further information on cells transfected with X-tremeGENE siRNA Transfection Reagent you can also browse the Transfection Database.


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Protocols and results provided by customers


Protocols shown here are provided by customers and can be obtained by clicking the cell type below. These protocols have not been verified by Roche scientists. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.

Note: Data and experimental conditions included in "Customer transfection protocols" are the sole responsibility of the customers that have submitted them. Roche was neither involved in establishing the experimental conditions nor in defining the criteria for the performance of the specific assays. Roche therefore cannot take any responsibility for performance or interpretation of results obtained for the biological target parameter(s) described by the authors or other users using a similar experimental approach.


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X-tremeGENE HP DNA Transfection Reagent


Cell name
(Click to view protocol)

Cell type

Transfected reporter gene

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

Not indicated

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

GFP

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

EGFP-CB2

293 (ATCC® CRL-1573™)

Human embryonic kidney cell line

Stable transfection

293-A

Human embryonic kidney cell line

EGFP fusion protein

293FT

Human embryonic kidney cell line subclone

GFP

293FT

Human embryonic kidney cell line subclone

Luciferase

293FT

Human embryonic kidney cell line subclone

Pelota/Ubc9

293T

Human embryonic kidney cell line expressing SV40 large T antigen

GFP

293T

Human embryonic kidney cell line expressing SV40 large T antigen

Interleukin /Fc fusion protein

3T3 (ATCC®CRL-1658™)

Mouse embryonic fibroblasts

GFP

A549 (ATCC®CCL-185™)

Human carcinomic alveolar basal epithelial cells

EGFP

AGS (ATCC®CRL-1739™)

Human epithelial adenocarcinoma cells

GFP

C2C12 (ATCC®CRL-1772™)

Mouse myoblast cell line

EGFP

CHO/dhFr- (ATCC® CRL-9096™)

Chinese hamster ovary cell line subclone

Secretable protein

CHO-K1 (ATCC® CCL-61™)

Chinese hamster ovary cell line subclone

GFP

COS-7 (ATCC® CRL-1651™)

African green monkey kidney fibroblast-like cell line

GFP

Dental papilla mesenchymal cells

Primary cells from the first lower molar of E18.5 Swiss mice

GFP

EC109

Esophageal squamous carcinoma cell line

Not indicated

EC9706

Esophageal squamous carcinoma cell line

Not indicated

F9 (ATCC® CRL-1720™)

Mouse embryonal carcinoma cells

EGFP

F9 (ATCC® CRL-1720™)

Mouse embryonal carcinoma cells

GFP

F9 (ATCC® CRL-1720™)

Mouse embryonal carcinoma cells

GFP

HCT 116 (ATCC® CCL-™)

Human colorectal carcinoma cell line

Not indicated

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

EGFP fusion protein

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

GFP and shRNA

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

HBV genome

HepG2.2.15

Human hepatoma cell line expressing complete HBV genome

GFP

HT-1080 (stably transfected)

Human fibrosarcoma cell line

EGFP-120 kDa nuclear protein

HuH-7 (ATCC® JCRB-0403)

Human hepatoma cell line

Hepatitis B surface antigen

HuH-7 (ATCC® JCRB-0403)

Human hepatoma cell line

Luciferase

HuH-7.5

Human hepatoma cell line

EGFP

L02

Immortal hepatic cell line

RNAi plasmid encoding GFP

LNCaP (ATCC® CRL-1740™)

Human prostate cancer cell line

GFP

MCF7 (ATCC® HTB-22™)

Human adenocarcinoma cell line

HA-tagged protein

MCF7 (ATCC® HTB-22™)

Human adenocarcinoma cell line

Not indicated

MEF (ATCC® HTB-22™)

Primary mouse embryonic fibroblasts

Not indicated

Plat-E

Human kidney packaging cell line

GFP

PK-15 (ATCC®CCL-33™)

Porcine kidney epithelial cell line

GFP

Porcine ovarian granulosa cells

Primary cells

EGFP

Stems cells, mesenchymal

Murine mesenchymal stem cells (RM26/2)

Far-red fluorescent protein

Stems cells, mesenchymal

Murine mesenchymal stem cells (EK8)

Far-red fluorescent protein

Stems cells, mesenchymal

Murine mesenchymal stem cells (p53-/-)

Far-red fluorescent protein

U-2 OS (ATCC® HTB-96™)

Human osteosarcoma cell line

GFP

U-87 MG (ATCC® HTB-14™)

Human epithelial glioblastoma cells

GFP


For further information on cells transfected with X-tremeGENE HP DNA Transfection Reagent you can also browse the Transfection Database.


Back to Top



X-tremeGENE 9 DNA Transfection Reagent


Cell name
(Click to view protocol)

Cell type

Transfected reporter gene

293-A

Human embryonic kidney cell line

EGFP

293FT

Human embryonic kidney cell line subclone

Luciferase

293T

Human embryonic kidney cell line

Luciferase

293T

Human embryonic kidney cell line

EGFP

293T

Human embryonic kidney cell line

Flag-tagged protein

BF-2 (ATCC® CCL-91™)

Bluegill fry (fish cell line)

RFP and EGFP

C2C12 (ATCC® CRL-1772™)

Mouse myoblast cell line

EGFP

CHSE-214 (ATCC® CRL-1681™)

Chinook salmon embryo (fish cell line)

EGFP

COS-7 (ATCC® CRL-1651™)

African green monkey kidney fibroblast-like cell line

EGFP-fusion protein

COS-7 (ATCC® CRL-1651™)

African green monkey kidney fibroblast-like cell line

GFP

EPC (ATCC® CRL-2872™)

Fish cell line (Epithelioma papulosum cyprini)

RFP

GF

Fish cell line (grouper fin)

RFP and EGFP

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

EGFP-actin

HeLa (ATCC® CCL-2™)

Human cervix carcinoma cell line

GFP

HepG2 (ATCC® HB-8065™)

Human hepatoma cell line

14 kb plasmid (incl. eGFP; stable)

HINAE

Japanese flounder (Hirame Natural Embryo) fish cell line

RFP and EGFP

HuH-7 (ATCC® JCRB-0403)

Human hepatoma cell line

Luciferase

MCF7 (ATCC® HTB-22™)

Human adenocarcinoma cell line

FAK-enconding shRNA plasmid

Neuro-2a (ATCC® CCL-131™)

Mouse neuroblastoma cells

GFP-mortalin fusion protein

PC-12 (ATCC® CRL-1721™)

Rat adrenal pheochromocytoma cell line

EGFP fusion protein

Primary pulmonary fibroblasts

Human pulmonary fibroblasts (primary)

Not indicated

Porcine granulosa cells

Porcine granulosa cells (subculture of primary cells)

GFP fusion protein

Sf9 (ATCC® TIB-71™)

Epithelial fall armyworm (Spodoptera frugiperda) cells

GFP


For further information on cells transfected with X-tremeGENE 9 DNA Transfection Reagent you can also browse the Transfection Database.


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