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Tau RD P301S FRET Biosensor

货号 TS204181
中文名称 null
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产品名称: Tau RD P301S FRET Biosensor
商品货号: TS204181
Organism: Homo sapiens, human
Tissue: embryonic kidney
Product Format: frozen 1.0 mL
Morphology: epithelial
Culture Properties: adherent
Biosafety Level: 2 xa0Cells contain SV40 and CMV 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
Gender: female
Applications: This cell line has been engineered to report tau seeding activity. Tau seeds introduced into the culture media of CRL-3275 can nucleate the aggregation of the endogenous tau reporter proteins. This aggregation produces a FRET signal which can be measured via microscopy, microplate readers or flow cytometry.
Images: Cell Micrograph of Tau RD P301S FRET Biosensor TS204181
Derivation:

The Tau RD P301S FRET Biosensor cells were derived by transducing HEK293T cells with 2 separate lentivirus constructs encoding tau RD P301S-CFP and tau RD P301S-YFP. Dual-positive cells were identified by FAC sorting and were cloned and isolated using cloning cylinders.

Complete Growth Medium: The base medium for this cell line is ATCC-formulated Dulbeccos Modified Eagles Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • 2mM L-alanyl-L-glutamine
  • fetal bovine serum to a final concentration of 10%

Subculturing:

Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Add 2.0 to 3.0 ml of 0.25% (w/v) Trypsin - 0.53 mM EDTA (ATCC 30-2101) solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

xa0xa0 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.

  1. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  2. Add appropriate aliquots of the cell suspension to new culture vessels.
  3. Incubate cultures at 37°C.

Subcultivation Ratio: 1:6 to 1:12 is recommended

Cryopreservation: Freeze Medium: Fetal Bovine Serum, 90%; DMSO, 10%
Storage Temperature: liquid nitrogen vapor phase
Culture Conditions: Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Volume: 1.0 mL
STR Profile: Amelogenin: X
CSF1PO: 11, 12
D13S317: 12, 14
D16S539: 9, 13
D5S818: 8, 9
D7S820: 11
TH01: 7, 9.3
TPOX: 11
vWA: 16, 19
COI:

Human

Name of Depositor: M. Diamond, University of Texas Southwestern Medical Center
Year of Origin: 2014
References:

Holmes BB, et al. Proteopathic tau seeding predicts tauopathy in vivo. Proc. Natl Acad. Sci. 111(41): E4376-4385, 2014. PubMed: 25261551

Hussain SA, et al. Fluorescence Resonance Energy Transfer (FRET) sensor. Scijet 4: 119, 2015 arXiv:1408.6559 physics.bio-ph