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GFPu-1 拉丁名

(ATCC? CRL-2794?) 統(tǒng)一編號(hào)

Organism Homo sapiens, human 

Tissue kidney 

Cell Type epithelialtransformed with adenovirus 5 DNA 

Product Format 提供形式 frozen 

Morphology epithelial Culture Properties adherent 

Biosafety Level 生物安全等級(jí) 2 

Cells containing Adenovirus viral DNA sequences 

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. 

Age fetus 

Applications 用途 The cell line can be used as a reporter of proteasome activity and enables the monitoring the function of the ubiquitin-proteasome system in an intact cell. 

This line was derived in July, 2000 from the human embryonic kidney line, 293. 293 cells were transfected with a reporter consisting of a short degron, CL1, fused to the COOH-terminus of green fluorescent protein (GFPu). 

Storage Conditions 保存方法 liquid nitrogen vapor phase

Derivation This line was derived in July, 2000 from the human embryonic kidney line, 293.293 cells were transfected with a reporter consisting of a short degron, CL1, fused to the COOH-terminus of green fluorescent protein (GFPu). 

The GFPu plasmid was created by ligating an oligonucleotide encoding ACKNWFSSLSHFVIHL into the GFP-C1 plasmid (Clontech) [PubMed:11375494]. 

Cells contain unstable green fluorescent protein (GFP). 

The cell line can be used as a reporter of proteasome activity and enables the monitoring the function of the ubiquitin-proteasome system in an intact cell. 

Ubiquitin dependent proteolysis has a role in cell division and apoptosis due to protein aggregation. 

Comments 注釋 This line was derived in July, 2000 from the human embryonic kidney line, 293.293 cells were transfected with a reporter consisting of a short degron, CL1, fused to the COOH-terminus of green fluorescent protein (GFPu). 

The GFPu plasmid was created by ligating an oligonucleotide encoding ACKNWFSSLSHFVIHL into the GFP-C1 plasmid (Clontech) [PubMed:11375494]. Cells contain unstable green fluorescent protein (GFP). 

The cell line can be used as a reporter of proteasome activity and enables the monitoring the function of the ubiquitin-proteasome system in an intact cell. 

Ubiquitin dependent proteolysis has a role in cell division and apoptosis due to protein aggregation.Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. 

To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. 

Subculturing Protocol: Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. 

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). 

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. 

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. 

An inoculum of 5 X 10(3) to 3 X 10(4) viable cells/cm2 is recommended. Maintain cultures at a cell concentration between 5 X 10(4) and 5 X 10(5) cells/cm2. Do not exceed 8 X 10(5) cells/cm2. Incubate cultures at 37°C. 

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended 

Medium Renewal: Two to three times weekly 

Cryopreservation Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO 

Storage temperature 保藏溫度 : liquid nitrogen vapor phase 

Culture Conditions 培養(yǎng)條件 

Atmosphere 需氧情況 : air, 95%; carbon dioxide (CO2), 5% 

Temperature 培養(yǎng)溫度 : 37.0°C 

Growth Conditions 生長(zhǎng)條件 : Do not allow cell density to exceed 90%. 

Population Doubling Time 17 hrs

Name of Depositor 寄存人 RR Kopito 

Year of Origin July, 2000 

References 參考文獻(xiàn) Bence NF, et al. Impairment of the ubiquitin-proteasome system by protein aggregation. Science 292: 1552-1555, 2001. PubMed: 11375494