Table 1 Substrate characteristics

From: Click beetle luciferase mutant and near infrared naphthyl-luciferins for improved bioluminescence imaging

 

Emission peaka (nm)

Brightnessb (RLU)

 

D-LH2

NH2-NpLH2

OH-NpLH2

D-LH2

NH2-NpLH2

OH-NpLH2

Luc2

559

678c, 719d

*

1.0

1 × 10–5

(−)

UltraGlo

556

555c, 667d

659

0.5

1 × 10−4

4 × 10−6

CBG99

546

544

*

1.0

2 × 10−2

2 × 10−6

CBR

614

664

758

0.2

2 × 10−4

3 × 10−6

  1. * Could not be obtained due to low signal. (−) undetectable signal
  2. a Average peak values (±3 nm) as determined from multiple (≥3) spectral reads
  3. b Calculated brightness data normalized to Luc2/D-LH2 (1.0); Reactions consisted of equal volumes of 1 µg mL−1 enzyme (in TBS (pH 7.5) containing 0.01% BSA) and 150 µM substrate+1 mM ATP (in 150 mM HEPES (pH 7.5), 1 mM CDTA, 16 mM MgSO4, and 1% NP-9); n = 3. Signal values for NH2-NpLH2 and OH-NpLH2 were lower than D-LH2 partly because of luminometer PMT bias for shorter wavelength photons
  4. c major peak
  5. d minor peak
Back to article page