BV2 cells (ATCC, Manassas, VA, USA, Cat# CRL-2467, RRID: CVCL_5744) were cultured with high-glucose Dulbecco’s Modified Eagle Medium (DMEM; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific) and high glucose DMEM/F12 (Thermo Fisher Scientific) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAFV600E, BRAFWT, and vector lentivirus, respectively, for 24 hours, cells were cultured for 24 hours in DMEM/F12 without fetal bovine serum (FBS), and then medium and cells were collected for subsequent experiments. Constructs used were: pMD2.G (Addgene, Cat# 12259, RRID: Addgene_12259), psPAX2 (Addgene, Cat# 12260, RRID: Addgene_12260), pHAGE-BRAFV600E plasmid (Addgene, Cat# 116204, RRID: Addgene_116204), pHAGE-BRAFWT plasmid (Addgene, Cat# 116719, RRID: Addgene_116719), pBabe-Puro-BRAFV600E plasmid (Addgene, Cat# 15269, RRID: Addgene_15269), gag/pol-Retroviral plasmid (Addgene, Cat# 14887, RRID: Addgene_14887), Control siRNA (Cell Signaling Technology, Boston, MA, USA, Cat# 6568S), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S).
Phage braf v600e plasmid
Manufactured by Addgene
Sourced in United States
The PHAGE-BRAF V600E plasmid is a bacterial expression vector that contains the BRAF V600E gene variant. The BRAF V600E mutation is a common genetic alteration found in various types of cancer. This plasmid can be used for the expression and study of the BRAF V600E protein in bacterial systems.
Automatically generated - may contain errors
Lab products found in correlation
Bv2 cells, by American Type Culture Collection (3 mentions)
Control sirna, by Cell Signaling Technology (3 mentions)
Erk2 sirna, by Cell Signaling Technology (3 mentions)
Phage braf wt plasmid, by Addgene (3 mentions)
Gag pol retroviral plasmid, by Addgene (3 mentions)
Erk1 sirna, by Cell Signaling Technology (3 mentions)
Pbabe puro braf v600e plasmid, by Addgene (3 mentions)
Sapk jnk sirna, by Cell Signaling Technology (3 mentions)
Pspax2, by Addgene (3 mentions)
Pmd2 g, by Addgene (3 mentions)
High glucose dmem f12, by Thermo Fisher Scientific (3 mentions)
High glucose dmem, by Thermo Fisher Scientific (2 mentions)
3 protocols using phage braf v600e plasmid
1
BV2 Cell Culture and BRAF Transduction
2
Lentiviral Transduction of BRAF Variants
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Constructs used were: pMD2.G (Addgene, Cat# 12259, USA), psPAX2 (Addgene, Cat# 12260, USA), pHAGE-BRAF V600E plasmid (Addgene, Cat# 116204, USA), pHAGE-BRAF WT plasmid (Addgene, Cat# 116719, USA), pBabe-Puro-BRAF V600E plasmid (Addgene, Cat# 15269, USA) , gag/pol-Retroviral plasmid (Addgene, Cat# 14887, USA), Control siRNA (Cell Signaling Technology, Cat# 6568S, USA), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S, USA), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S, USA), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S, USA). Microglial cell line BV2 cells BV2 cells (ATCC, Cat# CRL-2467, USA) were cultured with high-glucose DMEM (Gibco) containing 10% FBS (Gibco) and high glucose DMEM/F-12 (Gibco) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAF V600E , BRAF WT , and vector lentivirus, respectively, for 24 h, cells were cultured for 24 h in DMEM/F12 without FBS, and then medium and cells were collected for subsequent experiments.
3
Lentiviral Transduction of BRAF Variants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Constructs used were: pMD2.G (Addgene, Cat# 12259, USA), psPAX2 (Addgene, Cat# 12260, USA), pHAGE-BRAF V600E plasmid (Addgene, Cat# 116204, USA), pHAGE-BRAF WT plasmid (Addgene, Cat# 116719, USA), pBabe-Puro-BRAF V600E plasmid (Addgene, Cat# 15269, USA) , gag/pol-Retroviral plasmid (Addgene, Cat# 14887, USA), Control siRNA (Cell Signaling Technology, Cat# 6568S, USA), SAPK/JNK siRNA (Cell Signaling Technology, Cat# 6232S, USA), ERK1 siRNA (Cell Signaling Technology, Cat# 6436S, USA), ERK2 siRNA (Cell Signaling Technology, Cat# 6578S, USA). Microglial cell line BV2 cells BV2 cells (ATCC, Cat# CRL-2467, USA) were cultured with high-glucose DMEM (Gibco) containing 10% FBS (Gibco) and high glucose DMEM/F-12 (Gibco) containing 10% FBS at 37°C in 5% CO2 under constant temperature and humidity. After transduction with BRAF V600E , BRAF WT , and vector lentivirus, respectively, for 24 h, cells were cultured for 24 h in DMEM/F12 without FBS, and then medium and cells were collected for subsequent experiments.
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