Transcript CheLuminate

There is another top address in Darmstadt!
AppliChem CheLuminate
AppliChem © 2010
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There is another top address in Darmstadt!
 CheLuminate
• Chemiluminescence detection kits
• replaces radioactive assays
• extremely sensitive
AppliChem © 2010
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 CheLuminate
• Horseradish Peroxidase-driven
• Luminol-based
• plus enhancer
• New: plus nucleophilic acylation catalyst
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 CheLuminate
• Chemiluminescence detection kits
o for ELISA
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CheLuminate-HRP ELISA FemtoDetect
CheLuminate-HRP ELISA FemtoDetect Plus
o for highest sensitivity
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CheLuminate-HRP FemtoDetect
CheLuminate-HRP FemtoDetect Plus
CheLuminate-HRP PicoDetect Extended
o for standard applications
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CheLuminate-HRP PicoDetect
(with phenolic enhancer)
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 CheLuminate
• AppliChem's products:
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 CheLuminate
• Sensitivity and stability
Comparison of CheLuminate PicoDetect Extended with a competitor (B)
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 CheLuminate
• AppliChem CheLuminate – Sales arguments
o competitive price
o highest sensitivity available – from pico (10-12) to femto (10-15) range
o extremely high stability of reagents – up to 24 h signal duration
o compatible with most detection systems / instruments
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 CheLuminate
• Competitors
o Pierce (Thermo Scientific)
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Pierce ECL, SuperSignal™ West Pico,
SuperSignal™ West Femto
o Millipore
Immobilon™ Western HRP
o GE healthcare
ECL™
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 CheLuminate
• Literature
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 Bio-/Chemiluminescence
• Luciferase System
o Luciferase is one of the most sesnitive reporter in gene
expression studies; no radioactivity
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o No cellular background activity of luciferase in most
assay systems
o Easy to perform
o Linear activity over a wide range
APMBT
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 Bio-/Chemiluminescence
• Luciferase System with APMBT
o Luciferase from Photinus pyralis (Pluc)
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Luciferase from Renilla reniformis (Rluc)
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o application of both luciferases in the same assay possible:
internal control or investigation of two transcription signals
o first, measurement of Pluc, then Rluc under Rluc assay
conditions, Pluc inactive, because APMBT quenches Pluc
o This assay is compatible with protein assays!*
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APMBT = 2-(4-Aminophenyl)-6-methylbenzothiazole)
* Reference: Hampf, M. & Gossen, M. (2006) Anal. Biochem. 356, 94-99
comparable to Promega‘s DualGlow
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 Bio-/Chemiluminescence
• BCA
o Simple, sensitive photometric assay for proteins.
o Faster and easier than Lowry assay.
o With greater tolerance to interference from non-ionic
detergents,
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o and simple buffer salts. More stable at alkaline pH.
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The principle of the bicinchoninic acid (BCA) assay is similar to the Lowry procedure, in that both rely on the formation of a
Cu2+-protein complex under alkaline condition followed by reduction of the Cu2+ to Cu1+.
The amount of reduction is proportional to the protein present. In the second step, BCA forms a complex with Cu1+, which
is purple colored and is detectable at 562 nm.
Protein + Cu2+ => Cu1+ + BCA => Cu1+-BCA complex
market leader probably Pierce
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 Bio-/Chemiluminescence
• Overview
o substrates & inducers for
e.g. Cloning
Blue/White Selection:
X-Gal / IPTG
e.g. Histology
Alkaline Phosphatase Detection:
the detection of
• genes & proteins,
• enzyme activities
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BCIP / NBT
Diformazan
e.g. Western Blots
Horseradish Peroxidase Detection:
H2O2
Luminol
CheLuminate-HRP
Light
e.g. Reporter Genes
Firefly Luciferase Detection:
ATP
Luciferin
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Light
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