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L Wnt-5A
L Wnt-5A
規格:
價格:
編號:B206059
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
L Wnt-5A
商品貨號
B206059
Organism
Mus musculus, mouse
Tissue
subcutaneous connective tissue; areolar and adipose
培養基
Product Format
frozen
Morphology
fibroblast
Culture Properties
adherent
Biosafety Level
1

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.

Applications
They are presently the best source for production of Wnt-5A conditioned medium. Since the conditioned medium contains other factors besides the Wnt-5A protein, it is necessary to control any experiments involving the Wnt-5A conditioned medium with control conditioned medium from the parental cell line (ATCC CRL-2648).
Storage Conditions
liquid nitrogen vapor phase
Images
Derivation
L-M(TK-) cells (ATCC CCL-1.3) were transfected with a Wnt-5A expression vector and stable clones were selected in medium containing G418.
Comments

The Wnt-5A gene encodes a secreted glyoprotein with a variety of signaling effects. Wnt proteins bind to receptors of the Frizzled family on the cell surface. Wnt genes control many of the patterning and growth events during embryonic development. 

The cells secrete biologically active Wnt-5A protein. 

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: 0.4 mg/ml G-418; fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by pipetting gently.
  5. Add appropriate aliquots of cell suspension to new culture vessels. An inoculum of 6 x 103 to 8 x 103 viable cells/cm2 is recommended.
  6. Incubate culture vessels at 37°C. We recommend that you maintain cultures at a cell concentration between 5 x 104 and 3 x 105 cells/cm2.

Subcultivation Ratio: 1:4 to 1:6
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells: a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Population Doubling Time
36 hrs
Name of Depositor
R Nusse
References

Chen W, et al. Dishevelled 2 recruits beta-arrestin 2 to mediate Wnt5A-stimulated endocytosis of Frizzled 4. Science 301: 1391-1339, 2003. PubMed: 12958364

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