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Tau RD P301S FRET Biosensor
Tau RD P301S FRET Biosensor
規格:
價格:
編號:B172724
品牌:Mingzhoubio

標準菌株
定量菌液
DNA
RNA

規格:
凍干粉
斜面
甘油
平板


產品名稱
Tau RD P301S FRET Biosensor
商品貨號
B172724
Organism
Homo sapiens, human
Tissue
embryonic kidney
培養基
Product Format
frozen 1.0 mL
Morphology
epithelial
Culture Properties
adherent
Biosafety Level
2  [Cells 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 ATCC CRL-3275
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 Dulbecco's Modified Eagle's 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).

   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]

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