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Amplite® Reagents and Assay Kits

The Amplite® product series is composed of enzyme-based substrates and assays optimized for measuring horseradish peroxidase (HRP), luciferase, phosphodiesterases, proteases, histone deacetylase (HDAC) and cell signaling molecules such as NAD/NADH, NADP/NADPH, etc. The series includes assay kits spanning essentially all aspects of cellular and physiological activity, and is compatible with other AAT Bioquest products as well as classic materials.

 

 

Fluorogenic Substrates



Amplite® Red experimental results graph: HRP dose response was measured with Amplite® Fluorimetric Peroxidase Assay Kit in a black plate using a Gemini fluorescence microplate reader (Molecular Devices).
AAT Bioquest's substrates are a component of many of our kits, and are also available as standalone items. Spanning a wide spectral range, from the bright Amplite® Blue to the long-wavelength Amplite® IR, these substrates fluoresce in the presence of the peroxidase enzyme and H2O2. This series has a high signal-to-noise ratio and is very stable. Each substrate demonstrates useful features; Amplite® Blue does not photobleach, for example, and Amplite® IR has both excellent water solubility and is pH-independent from pH 4-10.
 

Table 1. Amplite® Substrates For Fluorogenic Substrates

Cat#
Product Name
Ex
Em
Unit Size
Price
11000Amplite® ADHP [10-Acetyl-3,7-dihydroxyphenoxazine] *CAS#: 119171-73-2*57158525 mg$205
11005Amplite® Blue32440925 mg$153
11009Amplite® IR6476701 mg$100
11011Amplite® Red5715851000 Assays$100

 

Quantitation Kits


Our Amplite® quantitation kits provide a rapid, dependable method of measuring a variety of biomaterials, ranging from alpha-ketoglutarate to zinc ions. Simple and efficient to use, these kits reliably measure the target materials, either in total amounts or by concentration, depending on the kit selected.

Assaywise Article Focus


Click the button on the right to read the related Assaywise article:
Or explore the entire Assaywise Newsletter archive starting with the most recent publication here.

 

Table 2. Amplite® Quantitation Kits For Organic Synthesis and Conjugation For Quantitation Kits


Table 3. Amplite® Quantitation Kits For Cell Metabolism and Oxidative Stress For Quantitation Kits


Table 4. Amplite® Quantitation Kits For Proteins in Solution For Quantitation Kits

Cat#
Product Name
Ex
Em
Unit Size
Price
11100Amplite® Fluorimetric Fluorescamine Protein Quantitation Kit *Blue Fluorescence*380464200 Tests$205
11105Amplite® Fluorimetric Protein Quantitation Kit *Orange Fluorescence*485590500 tests$205

Table 5. Amplite® Quantitation Kits For Ions For Quantitation Kits



Each of the Amplite® biomarker quantitation kits employ our proprietary Amplite® Red substrate, compatible with with common instrument platforms such as microplate or fluorescent microscope. A colorimetric format is also available for the detection of glucose.
 

Table 6. Amplite® Quantitation Kits For Biomarkers For Quantitation Kits

Cat#
Product Name
Ex
Em
Unit Size
Price
40001Amplite® Ethanol Quantitation Kit571585200 Tests$205
40004Amplite® Colorimetric Glucose Quantitation Kit575 500 Tests$205
40005Amplite® Fluorimetric Glucose Quantitation Kit571585500 Tests$205
40006Amplite® Cholesterol Quantitation Kit571585200 Tests$205
40007Amplite® Choline Quantitation Kit571585200 Tests$205

 

Endotoxin Detection Kits


Lipopolysaccharide, aka endotoxin, is a potentially-lethal biomarker of bacterial invasion. Its detection and prevention are of outmost importance in the pharmaceutical and biomedical industries. Employing Limulus Amebocyte Lysate (LAL), these endotoxin kits generate signal proportional to the presence of endotoxin in the sample. The kits are available in either colorimetric or fluorimetric format, and employ either Endotoxin Yellow™ or Endotoxin Green™ proprietary substrates, respectively. Both kits detect a wide range of endotoxin concentration, with the fluorimetric kit having approximately twice the sensitivity (detection of concentrations as low as 0.002 EU/ml) as the colorimetric kit.
 

Table 7. Amplite® Endotoxin Detection Kits For Endotoxin Detection Kits

Cat#
Product Name
Unit Size
Price
60006Amplite® Fluorimetric Endotoxin Detection Kit100 Tests$195
60007Amplite® Colorimetric Endotoxin Detection Kit100 Tests$195



Cytocite BG100 Portable Fluorometer

CytoCite™ Fluorometer Optimization


For researchers using the CytoCite™ or Qubit™ Fluorometers, we suggest using the Portelite™ Rapid Fluorimetric Endotoxin Detection Kit:
  • Also employs the components Endotoxin Green™ for hydrolyzation in the presence of endotoxins and the Limulus Amebocyte Lysate (LAL) to generate strong green fluorescence.


  • The kit is optimized for Cytocite™ and Qubit™ fluorometers, and can detect a broad range of endotoxin (from 1 EU/ml to 0.002EU/ml) present in the sample.


  • Very efficient: only requires 10 minutes incubation time.




 

Assay Kits



Enzyme Detection


Enzymes act as a regulatory catalyst for the majority of biochemical reactions that facilitate cell function, including all aspects of cellular metabolism. They do so by lowering the energy barrier necessary for a chemical reaction to occur. Enzymes are present throughout the cell, some sequestered within organelles such as the mitochondria, or free in the cytoplasm. Upon reaction with a suitable substrate, enzymes change shape to form a new complex. This change, when used during experiments, can provide a signal to researchers that a reaction has occurred, as well as potentially improving the specificity of the reaction itself. Amplite® enzyme-based assays provide a sensitive and simple method for the quantitation of a diverse array of analytes, including dehydrogenases, kinases, and other enzyme groups.

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Enzyme Activity


Enzyme activity assays are the primary tool for studying enzyme kinetics and inhibition. AAT Bioquest offers a full line of enzymatic activity kits for targets including hydrolytic enzymes, proteases, and other enzyme families.

Downloadable Resource


Click the button on the right to view or download our PDF:
Or explore our Application Notes pertaining to enzymes and other topics here.

 


In addition to the kits in the table above, AAT Bioquest offers a range of kits specifically for investigating the activity of caspases. Click the button below to learn more:

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Luciferase Reporter Gene



Luciferase activity was measured with Amplite® Luciferase Reporter Gene Assay Kit in a white 96-well plate with a NOVOstar plate reader (BMG Labtech). The luciferase activity is still detectable at 0.1 pg/well with 20 minutes incubation.
The luciferase reporter gene, originally isolated from the common eastern firefly Photinus pyralis, is the most popular choice for detection assays and other uses due to its low toxicity and ease of use. Our Amplite® kits employ different variations of this useful gene, isolated from the firefly, sea pansy (Renilla reniformis) and aquatic copepod Gaussia prince for sensitive, efficient detection and steady signal. AAT Bioquest also offers its bioluminescent substrate Luciferin in several forms.
 


Horseradish Peroxidase (HRP)


The peroxidase enzyme family plays a prominent role in multiple cellular processes. They catalyze hydrogen peroxide (H2O2) oxidation of a variety of organic compounds, many of which are commonly present in living cells. For use in the laboratory, the most commonly used one is isolated from the common horseradish. Horseradish peroxidase (HRP) is a small, robust molecule of only 44kD, and can be easily conjugated to antibodies in a 4:1 ratio with little to no steric hindrance. Since H202 is a common product of many cellular pathways, the ability to quantify it makes HRP incredibly valuable for multiple biochemical applications.

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Table 10. Amplite® HRP Assay Kits For Assay Kits



Alkaline Phosphatase (ALP)


Alkaline phosphatase (ALP) is an 86 kDa homodimeric protein enzyme present in multiple tissue types within the body, with the highest concentrations in the bone and liver. It is used as a biomarker for several pathologies. ALP is also a common reporter enzyme used in multiple immunoassays and other applications. It is typically conjugated to an antibody, and can be detected in multiple formats. The substrates for the Amplite® ALP kits range from the popular para-Nitrophenylphosphate (pnPP) for colorimetric detection and 4-Methylumbelliferyl phosphate (MUP) for fluorimetric, to our proprietary substrate for luminometric platforms.

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The application of Signal Guard™ phosphatase reaction stopping solution in Amplite® Alkaline Phosphatase Colorimetric Assay (AAT Bioquest Cat#11950). Two parallel reactions containing 100 mU/mL Alkaline phosphatase were initiated by adding 200 ?L reaction mixture. The plots demonstrate that the reaction was completely inhibited by Signal Guard™ phosphatase reaction stopping solution.

Phosphatase Assays and Reaction Control


For researchers requiring control over the phosphatase enzyme reaction, we suggest using the paired Signal Guard™ Phosphatase Reaction Stopping Solution:
  • The Signal Guard™ phosphatase reaction stopping solution is a ready to use reagent that provides a convenient tool for terminating fluorescence and colorimetric signal-generating phosphatase reactions at a user-determined time point, and also keep the fluorescence or colorimetric signal stable for up to 18 hours.


  • The stopping solution is optimized and compatible with alkaline, acid and protein phosphatase.


  • Compatible with nearly all colorimetric and fluorimetric phosphatase substrates, the solution requires only mild pH and only a 2-step protocol. Full details and pdf protocol available here.








NAD(P)(H) Assays


Nicotinamide adenine dinucleotides are prolific cofactors that facilitate the reversible oxidation and reduction steps in all major metabolic pathways. They are present in varying concentrations in phosphorylated (NADP and NADPH) and unphosphorylated forms (NAD+ and NADH+). Since the NAD(P)(H) cofactors link cell metabolism to other systems in the body (including transcriptional control and cell signaling) they are a target for drug discovery and oncology research. By monitoring the changes in intracellular concentration of these biomolecules, researchers gain understanding of cell behavior and pathologies.

Learn More

 

Table 12. Amplite® NAD/NADH Assay Kits For Assay Kits


Table 13. Amplite® NADP/NADPH Assay Kits For Assay Kits



Cellular Energy and Metabolism


All living cells must be able to fuel themselves with some form of energy. The process of transforming outside sources of energy (e.g. sunlight, food) into adenosine triphosphate (ATP) is the beginning of all life processes. Major metabolic pathways such as glycolysis are heavily regulated and include multiple enzymes and molecular intermediates. Assays to quantify changes in the concentrations of these biomolecules give valuable information about pathological mechanisms as well as normal cell activities.

Downloadable Resource


Click the button on the right to view or download our PDF:
Or explore our digital catalog page for cell metabolism here.

 

Table 14. Amplite® Metabolic Assay Kits For Assay Kits



Antioxidants and ROS


Reactive oxygen species (ROS) are a major component of cell signaling, gene expression, and other activities, with most occurring in the mitochondria. The amount of ROS in the cell is usually at low concentrations, as they are used for multiple cell signaling activities and other purposes. The levels are also kept low by the presence of antioxidants, which are synthesized or imported by the cell. In high concentrations however, these chemically-reactive molecules can be very damaging and even lethal.

Being able to accurately measure ROS levels is a useful way to detect the early stages of multiple pathologies, as well as the activity of metabolites or normal/atypical cellular function.
 


In addition to the kits in the table above, AAT Bioquest offers a range of kits specifically for investigating the activity of Glutathione (GSH and GSSG). Click the button below to learn more:

Learn More


Amino Acids


As the building blocks of all proteins, amino acids play a role in every aspect of cellular activity. Sensitive, accurate measurements of these small compounds, both essential and non-essential, are relevant in multiple fields of study.
 

Table 16. Amplite® Amino Acid Assay Kits For Assay Kits

Cat#
Product Name
Ex
Em
Unit Size
Price
10054Amplite® Fluorimetric Glutamic Acid Assay Kit *Red Fluorescence*571585200 Tests$205
13825Amplite® Fluorimetric L-Alanine Assay Kit 571585200 Tests$310
13826Amplite® Colorimetric L-Alanine Assay Kit 575 200 Tests$310
13827Amplite® Fluorimetric L-Aspartate (Aspartic Acid) Assay Kit 571585200 Tests$363
13828Amplite® Colorimetric L-Aspartate (Aspartic Acid) Assay Kit 575 200 Tests$363


Nerve Signaling


Sphingomyelin and acetylcholine, along with other roles, are important to signal transduction in the central nervous system. Using our proprietary Amplite® fluorescent substrates, these kits provide optimized protocols to give accurate, sensitive biomolecule quantification.

AAT Bioquest also offers Amplite® assay kits for the detection of the enzymes associated with these biomolecules, namely acetylcholinesterase (AChE) and sphingomyelinase (SMase), in a several formats and colors. Acetylcholinesterase (AChE) and sphingomyelinase (SMase) assays are one of the most popular methods for investigating nerve response and function. For more in-depth information on these critical tests see below:
 


Diagnostic Biomarkers


Specific biomolecules have distinctive changes in concentrations as a symptom of some pathologies. By measuring the changing levels of these materials against known standards, researchers gain insight into physiological conditions. Our Amplite® kits provide sensitive quantification with a minimal sample size of only 100 µL reaction volume.
 

Table 18. Amplite® Biomarker Assay Kits For Asssay Kits

Cat#
Product Name
Ex
Em
Unit Size
Price
11310Amplite® Fluorimetric Melanin Assay Kit470550100 Tests$310
13830Amplite® Colorimetric Beta-Hydroxybutyrate (Ketone Body) Assay Kit575 200 Tests$363
13831Amplite® Fluorimetric Beta-Hydroxybutyrate (Ketone Body) Assay Kit571585200 Tests$363
13832Amplite® Colorimetric Glycerol Assay Kit575 200 Tests$363
13833Amplite® Fluorimetric Glycerol Assay Kit571585200 Tests$363
13842Amplite® Colorimetric Xanthine Assay Kit575 200 Tests$363
13843Amplite® Fluorimetric Xanthine Assay Kit571585200 Tests$363

 

Product Ordering Information


 

Table 19. Product Ordering Information For Maleimide Quantification

Cat#
Product Name
Ex
Em
Unit Size
Price
5522Amplite® Colorimetric Biotin Quantitation Kit500 200 Tests$205
5523Amplite® Fluorimetric Maleimide Quantitation Kit *Green Fluorescence*490515200 Tests$205
5524Amplite® Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*510524200 Tests$205
5525Amplite® Colorimetric Maleimide Quantitation Kit324 100 tests$205
5526Amplite® Rapid Colorimetric Maleimide Quantitation Kit780 2 Tests$205
5528Amplite® Rapid Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*510524200 Tests$205
5529Amplite® Rapid Colorimetric Total Protein Thiol Quantitation Assay Kit680 2 Tests$205
10051Amplite® Colorimetric Aldehyde Quantitation Kit550 200 Tests$310
10052Amplite® Fluorimetric Aldehyde Quantitation Kit360450200 Tests$205
10053Amplite® Colorimetric Aldehyde Quantitation Kit *Blue Color*620 200 Tests$310