HEK293A GFP-LC3

14050801, human kidney, Small slightly rounded cells growing in a dispersed manner

• 分類程式碼代碼: 41106514

属性

• product name: HEK293A GFP-LC3, 14050801
• 生物源: human kidney
• 描述: Keywords: Human embryonic kidney, autophagy, HEK293, EGFP-rat LC3, reporter cell line, ATG8, microtubule-associated protein 1 light chain 3 beta (MAP1LC3B)
• 增長模式: Adherent
• 形態學: Small slightly rounded cells growing in a dispersed manner
• 技術: cell culture | mammalian: suitable
• 運輸包裝: dry ice
• 儲存溫度: −196°C

说明

細胞系描述

A stable cell line expressing EGFP-tagged LC3 (rat) in background of HEK293 cell line. This reporter cell line can be used to monitor induction of autophagy after amino acid starvation or rapamycin treatment as well as autophagosome formation. Also, this cell line provides a useful tool for the study of autophagy using GFP-LC3 as a standard assay read out by both immunofluorescence and biochemical methods.

To generate the cell line EGFP-LC3 (rat) plasmid was transfected into HEK293A cells. The cells were selected with 800ug/ml Geneticin. Single cell clones were selected and grown up. Several clones were tested and clone 2GL9 was taken as the best based on a number of parameters (including GFP-LC3 expression levels and responsiveness to amino acid deprivation).

Description of the genetic modification to generate this cell line: HEK293 cells transfected with a GFP-LC3 (green fluorescent protein-microtubule-associated protein 1 light chain 3) plasmid. cDNA encoding rat LC3 was obtained by RT–PCR from rat brain total cDNA and subcloned into the EcoRI site of the eukaryotic expression vector pCI-neo (Promega, Madison, WI). It was then excised and the LC3 cDNA inserted into the BglII and EcoRI sites of pEGFP-C1, a GFP fusion protein expression vector (Clontech Laboratories), Kabeya Y, et.al. (2000).

培養基

Dulbecco′s Modified Eagle′s Medium, high glucose (DMEM) + 2mM Glutamine + 10% Fetal Bovine Serum (FBS).

Geneticin at (400ug/ml) can be added for maintenance but this is not normally necessary as the GFP-LC3 expression level is maintained over at least 20 passages.

例行更新培養

Trypsinise cultures at 80% confluence with 0.05% Trypsin/EDTA. Seed flasks at 2 x 10⁴ cells/ cm². Cultures must be incubated in a humidified 10% CO₂/90% air incubator at 37^oC.

其他說明

Additional freight & handling charges may be applicable for Asia-Pacific shipments. Please check with your local Customer Service representative for more information.

Cultures from PHE Culture Collections and supplied by Sigma are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.

免責聲明

This cell line has special release conditions: Commercial organisations are required to complete the ′Cell Line Release Authorisation for Research Use in Commercial Organisations′ release conditions form.