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LanthaScreen™ TR-FRET CAR Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15141:

The LanthaScreen® TR-FRET Constitutive Androstane Receptor (CAR) Coactivator Assay provides a sensitive and robust method for highthroughput screening (HTS) of potential CAR ligands as agonists of ligand-dependent coactivator recruitment or inverse agonists of liganddependent coactivator displacement. The kit uses a terbium-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a CAR ligand-binding domain (CAR-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET CAR Coactivator Assay, CAR-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and terbium anti-GST antibody. After an incubation period at room temperature, the 520 nm/ 495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the CAR-coactivator peptide interaction, this ligand’s EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and either recruit or displace coactivator peptide (Figures 1 and 2).

Predictor™ hERG Fluorescence Polarization Assay Kit

The Predictor™ hERG Fluorescence Polarization Assay Kit provides a set of validated components to perform hERG channel biochemical binding studies in the absence of radioligand. The assay was designed to identify potential hERG channel blockers by producing data that accurately correlates with patch-clamp electrophysiology studies. The assay is based on the principle of fluorescence polarization where a red-shifted fluorescent tracer is displaced from the hERG channel by compounds that bind to the channel. Assay performance was validated with a panel of established hERG channel blockers. Results from the Predictor™ assay demonstrate a high correlation with those obtained from patch clamp techniques.

LanthaScreen™ TR-FRET PXR (SXR) Competitive Binding Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15142:

The LanthaScreen® TR-FRET PXR (SXR) Competitive Binding Assay Kit provides a sensitive and robust method for high-throughput screening (HTS) of ligands for the pregnane X receptor (PXR), also known as the steroid and xenobiotic receptor (SXR). The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescent small-molecule PXR ligand (Fluormone™ PXR (SXR) Green), and a human PXR (SXR) ligand-binding domain that is tagged with glutathione S-transferase (GST) in a homogeneous mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET PXR (SXR) competitive binding assay, a Tb-labeled anti-GST antibody is used to indirectly label the receptor by binding to its GST tag. Competitive ligand binding is detected by a test compound’s ability to displace a fluorescent ligand (tracer) from the receptor, which results in a loss of FRET signal between the Tb-anti-GST antibody and the tracer (Figure 1). This type of binding assay is analogous to radioligand-based assays, except that it eliminates handling of radioactivity and enables a homogeneous, “addition-only " format.

PolarScreen™ ER Alpha Competitor Assay, Red

The PolarScreen™ ER (Estrogen Receptor) Alpha Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential ER alpha ligands. The kit uses insect cell-expressed, full-length, native (untagged) human estrogen receptor alpha (available separately, Cat. No. A15674) and a novel, high-affinity, fluorescent estrogen ligand (Fluormone EL Red) in a homogenous, mix-and-read assay format.

This kit produces a red color read-out; a 'green' kit is also available. Both kits suitably determine the IC50s of competitive compounds. Color choice depends on the spectral properties of the compounds to be screened and the available instrumentation. For additional information or assistance contact drugdiscoverytech@lifetech.com.

PolarScreen™ ER Alpha Competitor Assay, Red, is:

• Complete & ready-to-use out of the box—all reagents included, just add your test compounds
• High throughput compatible—800 x 20 µL fluorescence polarization assays in a 384-well plate
• Convenient—available in bulk sizes for larger screening needs

How the Assay Works
Full length ER alpha is added to a fluorescent estrogen ligand to form an ER-Fluormone EL complex. This complex is added to individual test compounds in multiwell plates. To determine relative affinity, the assay measures fluorescence polarization. Competing test compounds will displace the Fluormone EL ligand from ER alpha, permitting it to tumble rapidly and resulting in low polarization values. A shift in the fluorescence polarization value in the presence of test compound is used to determine the relative affinity of test compounds for ER alpha.

All components are manufactured under the strictest quality parameters. This product's performance, per the Certificate of Analysis, is guaranteed for 6 months after purchase. Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

LanthaScreen™ TR-FRET PPAR alpha Competitive Binding Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15143:

The LanthaScreen® TR-FRET PPAR alpha Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor alpha (PPAR alpha). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and a human PPAR alpha ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST), in a homogeneous mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET PPAR alpha Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds followed by addition of a mixture of the PPAR alpha-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR alpha, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR alpha is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligandbased assays, except that it eliminates handling of radioactivity and enables a homogeneous "addition-only" format.

Urea Nitrogen (BUN) Colorimetric Detection Kit Invitrogen™

The Urea Nitrogen (BUN) Detection research-use-only kit is a colorimetric assay designed for the quantification and detection of urea nitrogen in serum, plasma, urine, saliva and tissue culture media samples.

This complete, ready-to-use kit includes clear 96-well plate(s), urea nitrogen standard, and other components to perform the assay. A 96-well microplate reader capable of reading optical absorption at 450 nm is required for use of this kit.

Performance characteristics
• Assay type: colorimetric detection kit
• Sample types: serum, plasma, urine, saliva, and tissue culture media samples
• Sensitivity: 0.030 mg/dL
• Standard curve range: 0.156 mg/dL–10 mg/dL
• Reactivity: species independent

Background
Urea is a by-product of protein metabolism by the liver, and is therefore removed from the blood by the kidneys. Urea freely filters through the glomerulous, but is reabsorbed by the renal tubules in a flow-dependent fashion. The higher the flow rate, the greater amount of urea nitrogen is cleared from circulation and eliminated through the kidneys. Urea nitrogen is identical across all species and this kit will measure urea nitrogen from sources other than human.

Assay principle
The Urea Nitrogen ((also called BUN) Colorimetric Detection Kit is designed to quantitatively measure urea nitrogen in a variety of samples. A urea nitrogen standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Samples are mixed with color reagents A and B and incubated at room temperature for 30 minutes. The colored product is read at 450 nm.

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LanthaScreen™ TR-FRET ER alpha Coactivator Assay Kit

The LanthaScreen® TR-FRET ER alpha Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential estrogen receptor (ER) alpha ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses an ER alpha ligand-binding domain (ER alpha-LBD) tagged with glutathione-S-transferase (GST), a terbium (Tb)-labeled anti-GST antibody, and a fluorescein-labeled coactivator peptide in a homogenous mix-and-read assay format.

Agonist Mode
To run the LanthaScreen TR-FRET ER alpha Coactivator Assay in agonist mode (to identify agonist compounds), ER alpha-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-anti-GST antibody. After incubation at room temperature, the TR-FRET 520:495 nm emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the ER alpha-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide.

Antagonist Mode
To run the LanthaScreen TR-FRET ER alpha Coactivator Assay in antagonist mode (to identify antagonist compounds), ER alpha-LBD is added to ligand test compounds, followed by addition of a mixture of agonist, fluorescein-coactivator peptide, and terbium (Tb)-labeled anti-GST antibody. The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

LanthaScreen™ TR-FRET PPAR delta Competitive Binding Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15144:

The LanthaScreen® TR-FRET PPAR delta Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor delta (PPAR delta). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and a human PPAR delta ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST), in a homogeneous mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET PPAR delta Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds, followed by addition of a mixture of the PPAR delta-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR delta, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR delta is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligandbased assays, except that it eliminates handling of radioactivity and enables a homogeneous, "addition-only" format.

LanthaScreen™ TR-FRET VDR Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15127:

The LanthaScreen® TR-FRET Vitamin D Receptor (VDR) Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential VDR ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a VDR ligand-binding domain (VDR-LBD) that is tagged with glutathione-S-transferase (GST), in a homogenous mix-and-read assay format.

The assay
To run the LanthaScreen® TR-FRET VDR Coactivator Assay, VDR-LBD is added to ligand test compounds, followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based upon the biology of the VDRcoactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET VDR Coactivator Assay Kit contains the VDR-LBD (GST) protein, fluorescein-labeled TRAP220/DRIP-2 coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

EnzChek™ Pyrophosphate Assay Kit Invitrogen™

The EnzChek® Pyrophosphate Assay Kit provides a rapid and highly sensitive enzymatic assay for detecting free pyrophosphate in solution. In this assay the formation of a chromophoric product is detected spectrophotometrically.

See our complete line of Fluorescence Microplate assays.

• Sensitivity limit of 1 µM pyrophosphate (~0.2 µg/mL)
• Acommodates a pH range of 6.5 to 8.5

The EnzChek® Pyrophosphate Detection Kit provides a fast, convenient, and inexpensive spectrophotometric method for measuring the inorganic pyrophosphate produced by a number of biochemical reactions, such as DNA and RNA polymerizations, cyclic AMP formation by the enzyme adenylate cyclase, and the enzymatic activation of fatty acids to form their coenzyme A esters.

The EnzChek® Pyrophosphate Assay Kit is a modification of our EnzChek® Phosphate Assay Kit (Cat. No. E6646). In the presence of inorganic phosphate, the substrate 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG) is converted by the purine nucleoside phosphorylase (PNP) enzyme to ribose 1-phosphate and 2-amino-6-merc apto-7-methylpurine. The enzymatic conversion of MESG results in a shift in absorbance maximum from 330 nm to 360 nm. The EnzChek® Pyrophosphate Assay Kit includes the enzyme inorganic pyrophosphatase, which catalyzes conversion of pyrophosphate into two equivalents of phosphate. The phosphate is then consumed by the MESG/PNP reaction and detected by an increase in absorbance at 360 nm. Additional sensitivity is gained by the amplification of one molecule of pyrophosphate into two molecules of phosphate.

LanthaScreen™ TR-FRET ER alpha Competitive Binding Kit

The LanthaScreen® TR-FRET ER alpha Competitive Binding Kit provides a sensitive and robust method for high-throughput screening (HTS) of ligands for estrogen receptor (ER) alpha. The kit uses a human ER alpha ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST), a terbium-labeled anti-GST antibody, and a fluorescent small-molecule ER ligand (Fluormone™ ES2) in a homogeneous mix-and-read assay format.

The Lanthascreen ER alpha Competitive Binding Assay is:

• Complete & ready-to-use out of the box—all reagents included, just add your test compounds
• High throughput compatible—TR-FRET assays in a 384-well plate
• Convenient—components available in bulk sizes for larger screening needs

LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15133:

The LanthaScreen® TR-FRET PPAR (peroxisome proliferator activated receptor) alpha Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential PPAR alpha ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant PPAR alpha ligand-binding domain (PPAR alpha-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.

Agonist mode:
When running the LanthaScreen® TR-FRET PPAR alpha Coactivator Assay in agonist mode (to identify agonist compounds), PPAR alpha- LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR alpha-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (see Figure 1).

Antagonist mode:
When the LanthaScreen® TR-FRET PPAR alpha Coactivator Assay is run in antagonist mode (to identify antagonist compounds), PPAR alpha-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-labeled anti-GST antibody (Figure 2). The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

Contents and Storage:
The LanthaScreen® TR-FRET PPAR alpha Coactivator Assay Kit contains PPAR alpha-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or 4°C).

LanthaScreen™ TR-FRET PPAR gamma Competitive Binding Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15145:

The LanthaScreen® TR-FRET PPAR gamma Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor-gamma (PPAR gamma). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and human PPAR gamma ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST) in a homogeneous mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET PPAR gamma Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds followed by addition of a mixture of the PPAR gamma-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR gamma, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR gamma is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligand-based assays, except that it eliminates handling of radioactivity and enables a homogeneous, "addition-only" format.

LanthaScreen™ TR-FRET LXR beta Coactivator Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4658:

The LanthaScreen® TR-FRET LXR (liver X receptor) beta Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential LXR beta ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and an LXR beta ligand-binding domain (LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous mix-and-read assay format.

How it works:
In the LanthaScreen® TR-FRET LXR beta Coactivator Assay, LXR beta-LBD is added to ligand test compounds followed by the addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the LXR beta-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).

Contents and Storage:
The LanthaScreen® TR-FRET LXR beta Coactivator Assay Kit contains LXR beta-LBD (GST) protein, fluorescently labeled D22 coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).

Ferric Antioxidant Status Detection Kit Invitrogen™

The Ferric Antioxidant Status Detection research-use-only kit is a colorimetric assay designed for the quantification and detection of Ferric Antioxidant Status (also referred as Ferric Reducing Antioxidant Power, FRAP) in serum, plasma, urine, teas, fruit juices, beer, cider, cell lysates, herbal and fruit extracts.

This complete, ready-to-use kit includes clear 96-well plate(s), ferrous chloride standard, ascorbic acid control, and other components to perform the assay. A 96-well microplate reader capable of reading optical density at 560 nm is required for use of this kit.

Performance characteristics
• Assay type: colorimetric detection kit
• Sample types: serum, plasma, urine, teas, fruit juices, beer, cider, cell lysates, herbal and fruit extracts
• Sensitivity: 8.06 uM
• Standard curve range: 31.2 uM–1,000 uM
• Reactivity: species independent

Background
Reactive oxygen species (ROS) are produced as a consequence of normal aerobic metabolism. These free radicals (FR) are usually removed or converted into other products in vivo by an array of antioxidants. Organisms have developed complex antioxidant systems to protect themselves from oxidative stress, including the activity of specific enzymes (especially superoxide dismutase, glutathione peroxidase, catalase, glutathione reductase) as well as nonenzymatic compounds with antioxidant activity, such as beta-tocopherol, L-ascorbic acid, glutathione, coenzyme Q10, flavonoids, and albumin. The assay measures the antioxidant ability from all species.

Assay principle
The Ferric Antioxidant Status Detection kit is designed to quantitatively measure antioxidant status in a variety of samples. A ferrous chloride standard is provided to generate a standard curve for the assay and all samples should be read off of the standard curve. Samples are diluted in assay buffer and added to the wells. The FRAP color solution is made by mixing reagent A and B with assay buffer. The FRAP color solution is added to all wells and the plate incubated at room temperature. Antioxidant power in the samples reacts with the FRAP color solution to generate a blue-colored product which is read at 560 nm.

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