Reagents for Cell Study and Assays

Cell Biology Tools

reagents for cell study and assays

AMCH-biotin - BAPA - HSA - Coelenterazines - Luciferins

AMCH- biotin	UPR0756
Aldehyde reactive biotin specific for a basic sites of DNA (apoptosis detection)

BAPA	UP84961
Colorimetric assays for site-carboxyl containing enzymes, especially factor XII, and for cellular transglutaminases

HSA

UP54420

Used as a retrograde tracer for neurons and also for a histochemical stain.

Hydroxystilbamide

MW 473

l (pH7) = 361 nm

e(361 nm) =>16000

Coelenterazines

Introduction - Luminescentes propreties of Coelenterazines

Native Coelenterazine

Coelenterazine cp

Coelenterazine e

Coelenterazine f

Coelenterazine fcp

Coelenterazine h

Coelenterazine hcp

Coelenterazine ip

Coelenterazine n

Coelenterazine 2-methyl analog (methyl Coelenterazine)

Introduction

Uptima proposes 10 different coelenterazine analogs:

Substrates for the aequorin complex / Detection based on luminescence ratio (Coelenterazine e)
Antioxidant (Coelenterazine methyl)

Bioluminescence is the light produced in a biochemical reaction involving the oxidation of a substrate via an enzyme. This phenomenon bas been used extensively in different formats for life science research and drug discovery owing to its extremely high sensitivity and non hazardous nature.

Examples of bioluminescence applications include calcium detection in live cells or tissues, reporter gene assays, ELISA, bioluminescence resonance energy transfer (BRET) for protein interaction studies, superoxide anion detection and high throughput drug screening.

The enzymes involved in most of these applications are apoaequorin, Renilla luciferase, firefly luciferase or a combination of firefly luciferase with another enzyme such as alkaline phosphatase, b-galactosidase or b-glucuronidase.

The key enzyme substrates are native, cp, h, coelenterazines (for apoaequorin and Renilla luciferase) and D-luciferin (for firefly luciferase).

Luminescentes propreties of Coelenterazines (Table 1)

Cat #	Coelenterazine	Maximum Emission (nm)	Relative Luminescence Capacity	Relative Intensity	Half-Rise Time (s)
UP97233	native	465	1.00	1.00	0.4 - 0.8
UPR3079	cp	442	0.95	15	0.15 -0.3
UP43876	f	473	0.80	18	0.4 - 0.8
UPR4711	fcp	452	0.57	135	0.4 - 0.8
UPR3078	h	464	0.82	10	0.4 - 0.8
UP08353	hcp	444	0.67	190	0.15 -0.3
UPR4712	ip	441	0.54	47	1
UP39819	n	467	0.26	0.01	5
UPT86770	e	405 and 465	0.5	4	0.15 - 0.3
UPT8889	2-methyl	-	-	-	-

Native Coelenterazine	UP972332	1mg

C₂₆H₂₁N₃O₃ MW 414.40
Coelenterazine is the luminophore of the native aequorin complex and also the substrate for Renilla luciferase. Bioluminescent detection of calcium concentration is highly sensitive in a broad concentration range (0.1µM to >100µM) 1-4. Monitoring of reporter genes (phot gene and luc gene) using coelenterazine is also a major application. Other uses of coelenterazine include bioluminescence resonance energy transfer(BRET)5 and chemiluminescent detection of superoxide anion and peroxynitrite in cells or tissues6-9. Uptima offers 10 synthetic coelenterazine derivatives that allow users to choose the ones that best suit the specific applications. Yellow solid soluble in MeOH or EtOH. Luminophore of the aequorin complex which is oxidized by oxygen to illuminate at 465 nm when Ca2+ binds to the complex. See Table 1 for comparison
*Literature* Meth. Cell Biol. 40, 305(1994) Meth. Enzymol. 172, 164(1989) J. Cell Biol. 115, 1259(1991) Cell Calcium, 14, 373 (1993) Proc. Natl. Acad. Sci. USA 96, 151(1999) Free Radic. Biol. Med. 28, 1232(2000) Circ. Res. 84, 1203(1999) Immunol. Today 15, 7(1994) Anal. Biochem. 206, 273(1992)

Coelenterazine cp	UPR30792	50µg
	UPR30793	1mg
C₂₅H₂₅N₃O₃ MW 415.48
Coelenterazine cp is a synthetic derivative of coelenterazine. Its aequorin complex generates luminescence intensity 15 times higher and has a faster response time to calcium than native coelenterazine does. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine e NEW	UPT86770	Inquire

C₂₈H₂₃N₃O₃ MW 449.50
Coelenterazine e is a synthetic derivative of coelenterazine. Compared with coelenterazine (native form), aequorin formed from coelenterazine e has a faster luminescence rise time and shows two emission peaks at 405 and 465 nm respectively, with the ratio of the peak heights dependent on calcium concentration in the range of pCa 5-7. See Table 1 for comparison
*Literature* Biochem. J 251, 405(1988)

Coelenterazine f	UP438761	50µg
	UP438762	1mg

Coelenterazine f is a synthetic derivative of coelenterazine. The luminescence intensity of its aequorin complex is almost 20 times higher than that of native coelenterazine while its emission maximum is about 8 nm longer than that of the latter. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine fcp	UPR47110	50µg
	UPR47111	1mg
C₂₅H₂₄FN₃0₂ MW 417.48
Coelenterazine fcp is a synthetic derivative of coelenterazine. Its luminescence intensity is 135 times higher than that of native coelenterazine. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine h	UPR30782	50µg
	UPR30783	1mg
C₂₆H₂₁N₃0₂ MW 407
Coelenterazine h is a synthetic derivative of coelenterazine but its luminescence intensity is more than 10 times higher than that of the latter. See Table 1 for comparison

Coelenterazine hcp	UP083533	50µg
	UP083534	1mg
C₂₅H₂₅N₃O₂ MW 399.49
Coelenterazine hcp is a synthetic derivative of coelenterazine. The luminescence intensity of its aequorin complex is 190 times higher than that of aequorin complex formed from native coelenterazine while the response time to calcium is faster. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine ip	UPR47120	50µg
	UPR47121	1mg
C₂₃H₂₃N₃0₃ MW 389.45
Coelenterazine ip is a synthetic derivative of coelenterazine. Its luminescence intensity is almost 50 times higher than that of native coelenterazine while its response time to calcium is much slower than the latter. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine n	UP398192	50µg
	UP398193	1mg
C₃₀H₂₃N₃O₂ MW 457.52
Coelenterazine n is a synthetic derivative of coelenterazine. Its luminescence intensity is the weakest of all coelenterazine analogs and its response time to calcium is also much slower than that of native. Coelenterazine n is reported to be a very useful low-sensitivity coelenterazine. See Table 1 for comparison
*Literature* Biochem. J. 261, 913(1989) Cell Calcium 12, 635(1991) Cell Calcium, 14, 373 (1993)

Coelenterazine, 2-methyl	UPT88890	50µg
analog (methyl Coelenterazine)	UPT88891	1mg
C₂₀H₁₇N₃O₂ MW 331.37
Methyl coelenterazine has been reported to be a superior antioxidant for cells against reactive oxygen species (ROS) such as singlet oxygen and superoxide anion. The coelenterazine derivative is membrane-permeant, nontoxic and highly reactive toward ROS. As oxidative stress is believed to be a mediator o apoptosis, methyl coelenterazine should be another important tool for apoptosis study. It is also a potent antioxidant D-Luciferin free acid. See Table 1 for comparison
*Literature* Biochem. Pharmacol. 60, 471 (2000) Immunol. Today 15, 7 (1994) Anal. Biochem. 206, 273 (1992) Circ. Res. 84, 1203 (1999)

Luciferins

Introduction

D-Luciferin, free acid

D-Luciferin, K salt

D-Luciferin, Na salt

Introduction

D-Luciferin, a monomeric 61kDa protein, is a natural compound isolated from fireflies (Photinus pyralis) and other beetles and is a substrate for the enzyme luciferase. Our highly purified synthetic luciferin exhibits physical properties identical to those of natural luciferin
Thanksa to very brief duration of light output, optimized formulations allows linear results over at >8 orders of magnitude of enzyme concentration, down less than 10-20 moles of luciferase. Generally, 50/100-fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase (CAT) assay. A nearly constant signal for 1minute or more can be achieved detected in single-tube luminometer or in a multiwell plate luminometer with an autoinjector.
Applications: the luciferin/luciferase system is used as a very sensitive reporter assay for gene expression for plants, bacteria, mammalian cells, and for monitoring baculovirus gene expression in insects. It can too be used for ATP assays in research applications or to detect bacterial contamination. It also has been used for detecting certain amphipathic and hydrophobic substances, including anesthetics and hormones, as these compete with luciferin for the hydrophobic site on the luciferase molecule (Anal Biochem 190, 304 (1990) ).

Technical Sheet

D-Luciferin, free acid	27060A
(S)-4.5-Dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid C₁₁H₈N₂O₃S₂ CAS [2591-17-5] MW 280 Purity: >99%
D-Luciferin, K salt	M1224A
C₁₁H₇KN₂O₃S₃ CAS [ 115144-35-9] MW 318.42 Purity: >99%
D-Luciferin, Na salt	72604A
C₁₁H₇N₂NaO₃S₃ CAS [103404-75-7] MW 302.30 Purity: >99%

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