Planned maintenance

We must perform activities that will impact the ability to place orders on our website. Starting Friday, May 21, at 5:00 p.m. Eastern to Sunday, May 23, at 5:00 p.m. Eastern, you will not be able to order products on our website. You will still be able to browse atcc.org and access your account. We apologize for any inconvenience this work may cause.
X

Tau RD P301S FRET Biosensor (ATCC® CRL-3275)

Organism: Homo sapiens, human  /  Tissue: embryonic kidney  / 

Permits and Restrictions

View Permits View Restrictions

Organism Homo sapiens, human
Tissue embryonic kidney
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]

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • For-profit customers must obtain a research use license agreement from the Contributor prior to shipment.
  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. Prior to purchase, any for-profit Purchaser must obtain a research use license from Washington University; and
  2. When a Commercial Use is contemplated, all for-profit and non-profit Purchasers must obtain a Commercial Use license from the contributor.  For information on executing a license to use this product for research (for-profit entities) or Commercial Use (for-profit and non-profit entities), contact:
Washington University Office of Technology Management
Attn. Associate Director
E-mail: otm@wustl.edu
Phone: 314.747.0920.

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]