UptiBlue

UptiBlue Viable Cell Counting Reagent

Cell growth and cytotoxicity indicator

Use UptiBlue instead of MTT, XTT, [³H]-Thymidine or Neutral Red

• Water soluble

• Nontoxic to cells

• Nontoxic to user

• Stable in culture

• Fluorescent and colorful

• No extraction step required

• No metabolic interference

• Noncarcinogenic

• Allows continuous monitoring of cells

• Use any microplate spectrophotometer or fluoromete

UptiBlue is an indicator dye, formulated to measure quantitatively the proliferation of a variety of human or animal cells, bacteria,or fungi. It consists of an oxidation-reduction indicator that yields a colorimetric change and a fluorescent signal in response to metabolic

Courb showing increased absorbance and             fluorescence in proportion to greater cell density and metabolic activity.

Continuous culture of A549 cells in UptiBlue. Cells were grown in medium supplemented with FBS and UptiBlue. Increasing fluorescence reflects increasing cells number.

Phenol red and high concentrations of protein, such as those found in FBS, do not inhibit UptiBlue.

Comparaison of inhibition curves showing the LD₅₀ for 5-fluorouracil and doxorubicin in DU145 cells using UptiBlue or MTT.

• UptiBlue

• MTT

1. Expose cells to test agents
2. Add UptiBlue to all test wells
3.  Incubate for 3 hours at 37°C, 5% C0₂, >90% humidity

4. Read absorbance at 570 nm et 600 nm or measure fluorescence with excitation wavelenght at 560 nm, and emission wavelenght of 590 nm

1. Expose cells to test agents
2. Add MTT to all test wells
3.  Incubate for 3 hours at 37°C, 5% C0₂, >90% humidity Rotate or shake every 10-15 minutes during incubation
4. Aspirate and discard MTT solution
5. Suspension cell lines must be centrifuged at low speed for 5 mn prior to removal of the solution
6. Add DMSO to each well
7. Agitate plate until thorough formazan solubilization has occured (may require up to 15 minutes)
8. Read absorbance at 540 nm

Technical Sheet

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03103 Montluçon cedex - France

Tel. (33) (0) 4 70 03 88 55

Fax. (33) (0) 4 70 03 82 60

uptima@interchim.com

www.interchim.com