Shop All Biochemical Assay Kits

cAMP-Screen™ Cyclic AMP Immunoassay System (Applied Biosystems™)

The cAMP-Screen® 96-well Cyclic AMP Immunoassay System enables ultrasensitive determination of cyclic AMP (cAMP) levels in cell lysates. This competitive chemiluminescent immunoassay is formatted with maximum flexibility to permit either manual reagent additions or automated high-throughput processing. The cAMP assay utilizes the highly sensitive chemiluminescent CSPD® Substrate with Sapphire-II™ Enhancer that is triggered by an enzyme conjugate composed of cAMP bound to alkaline phosphatase (cAMP-AP). The chemiluminescent substrate/enhancer formulation is a ready-to-use reagent that generates a sustained-glow light emission from 30 minutes after addition. This assay is mainly used in secondary screening and pre-clinical research, where sensitivity and no false positives are essential.

• The highest sensitivity of any commercially available cAMP assay
• Hours of read time with little or no degradation of the signal
• A simple protocol
• Useful for a wide range of receptor activation studies

High Sensitivity and Hours of Steady Glow Time
This chemiluminescent assay is designed to provide the highest sensitivity of any commercially available cAMP assay. As few as 60 femtomoles of cAMP can be detected. The assay has a wide dynamic range, detecting from 0.06 to 6,000 picomoles of cAMP without the need for sample dilution or manipulations such as acetylation. This is especially helpful in cell-based assays when measuring Gs- or Gi-coupled agonist or antagonist stimulation and/or inhibition. Intra-assay precision for duplicate samples is typically 5% or less. Once the substrate/enhancer formulation reaches glow signal, the plate can be read for hours with little or no degradation of the signal. This is useful in screens where several plates are compared with each other. In addition, the assay exhibits exceptionally low cross-reactivity with other adenosine-containing or cyclic nucleotides.

For Receptor Activation Studies
The cAMP-Screen® system is designed for quantitation of cellular cAMP for functional assays of receptor activation. The assay has been used with established cell lines for functional measurements with endogenous receptors, cell lines with exogenously expressed ligand receptors on the cell surface, primary cell cultures, and tissues in response to treatment with the appropriate ligands. In addition it has been used for receptor characterization, orphan receptor ligand identification, and the characterization of novel chimeric receptors. The assay can be used for high-throughput screens for compounds that stimulate or interfere with these signal transduction pathways.

A Simple Protocol
The assay follows a simple protocol. Cells are seeded into plates, cultured, and treated with test compounds as desired. Cell lysates are prepared in either the presence or absence of culture media. Lysates are incubated with a cAMP-AP conjugate and an anti-cAMP antibody in a coated microplate; the resulting immune complexes are captured in the plate. In samples without cAMP, all of the cAMP-AP conjugate is captured on the coated surface, resulting in a high signal. In the presence of cAMP, the amount of cAMP-AP conjugate captured decreases as a result of competition for binding with unlabeled cAMP, causing a reduced signal; signal reduction is proportional to the amount of cAMP present in the cell lysate. After washing to remove unbound cAMP-AP, the chemiluminescent substrate is added, and the resulting glow signal is measured in a luminometer.

An alternate product, the cAMP-Screen Direct® system, has all of the same benefits as this cAMP-Screen® system, but eliminates the need to transfer cell lysates from a tissue culture plate to the precoated microplates because the cells can be grown directly in the microplates. This provides better assay precision with one less transfer step. Many different cell types have been grown successfully on the precoated microplates, which also have a clear bottom to allow monitoring of cells.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

cAMP-Screen Direct™ Cyclic AMP Immunoassay System (Applied Biosystems™)

The cAMP-Screen Direct® 96-well Cyclic AMP Immunoassay System enables ultrasensitive determination of cyclic AMP (cAMP) levels in cell lysates. This competitive chemiluminescent immunoassay is formatted with maximum flexibility to permit either manual reagent additions or automated high-throughput processing. The cAMP assay utilizes the highly sensitive chemiluminescent CSPD® Substrate with Sapphire-II™ Enhancer that is triggered by an enzyme conjugate composed of cAMP bound to alkaline phosphatase (cAMP-AP). The chemiluminescent substrate/enhancer formulation is a ready-to-use reagent that generates a sustained-glow light emission from 30 minutes after addition. This assay is mainly used in secondary screening and pre-clinical research, where sensitivity and no false positives are essential.

• The highest sensitivity of any commercially available cAMP assay
• Hours of read time with little or no degradation of the signal
• A simple protocol with cells grown directly on the assay plate
• Useful for a wide range of receptor activation studies

High Sensitivity and Hours of Steady Glow Time
This chemiluminescent assay is designed to provide the highest sensitivity of any commercially available cAMP assay. As few as 60 femtomoles of cAMP can be detected. The assay has a wide dynamic range, detecting from 0.06 to 6,000 picomoles of cAMP without the need for sample dilution or manipulations such as acetylation. This is especially helpful in cell-based assays when measuring Gs- or Gi-coupled agonist or antagonist stimulation and/or inhibition. Intra-assay precision for duplicate samples is typically 5% or less. Once the substrate/enhancer formulation reaches glow signal, the plate can be read for hours with little or no degradation of the signal. This is useful in screens where several plates are compared with each other. In addition, the assay exhibits exceptionally low cross-reactivity with other adenosine-containing or cyclic nucleotides.

For Receptor Activation Studies
The cAMP-Screen Direct® system is designed for quantitation of cellular cAMP for functional assays of receptor activation. The assay has been used with established cell lines for functional measurements with endogenous receptors, cell lines with exogenously expressed ligand receptors on the cell surface, primary cell cultures, and tissues in response to treatment with the appropriate ligands. In addition it has been used for receptor characterization, orphan receptor ligand identification, and the characterization of novel chimeric receptors. The assay can be used for high-throughput screens for compounds that stimulate or interfere with these signal transduction pathways.

A Simple Protocol—No Lysate Transfer Required
The assay follows a simple protocol. Cells are seeded into plates, cultured, and treated with test compounds as desired. Cell lysates are prepared in either the presence or absence of culture media. Lysates are incubated with a cAMP-AP conjugate and an anti-cAMP antibody in a coated microplate; the resulting immune complexes are captured in the plate. In samples without cAMP, all of the cAMP-AP conjugate is captured on the coated surface, resulting in a high signal. In the presence of cAMP, the amount of cAMP-AP conjugate captured decreases as a result of competition for binding with unlabeled cAMP, causing a reduced signal; signal reduction is proportional to the amount of cAMP present in the cell lysate. After washing to remove unbound cAMP-AP, the chemiluminescent substrate is added, and the resulting glow signal is measured in a luminometer.

This cAMP-Screen Direct® system has all of the same benefits as the cAMP-Screen® system, but also eliminates the need to transfer cell lysates from a tissue culture plate to the precoated microplates because the cells can be grown directly in the microplates. This provides better assay precision with one less transfer step. Many different cell types have been grown successfully on the precoated microplates, which also have a clear bottom to allow monitoring of cells.

For Research Use Only. Not for human or animal therapeutic or diagnostic use.

LanthaScreen™ TR-FRET Glucocorticoid Receptor Coactivator Assay Kit

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

Agonist Mode
When running the LanthaScreen TR-FRET Glucocorticoid Receptor Coactivator Assay in agonist mode (to identify agonist compounds), GR-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-labeled anti-GST antibody. After incubation at room temperature, the 520/495 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 GR-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).

Antagonist Mode
When the LanthaScreen TR-FRET Glucocorticoid Receptor Coactivator Assay is run in antagonist mode (to identify antagonist compounds), GR-LBD is added to ligand test compounds followed by the addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-labeled anti-GST antibody (see figure). 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 Glucocorticoid Receptor Competitive Binding Kit

The LanthaScreen® TR-FRET Glucocorticoid Receptor Competitive Binding Assay Kit provides a sensitive and robust method for high-throughput screening (HTS) of ligands for glucocorticoid receptor (GR). The kit uses a human GR ligand-binding domain (GR-LBD) tagged with glutathione S-transferase (GST) (also available separately), a terbium-labeled anti-GST antibody, and a fluorescent small-molecule GR ligand (Fluormone™ GS1) in a homogeneous mix-and-read assay format.

The Lanthascreen Gluococorticoid Receptor 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 Progesterone Receptor Coactivator Assay Kit

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

Agonist Mode
When using the LanthaScreen TR-FRET Progesterone Receptor Coactivator Assay in agonist mode (to identify agonist compounds), PR-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-labeled coactivator peptide and Tb-labeled anti-GST antibody. After an incubation period at room temperature, the TR-FRET ratio of 520/495 nm is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PR-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).

Antagonist Mode
When using the LanthaScreen™ TR-FRET Progesterone Receptor Coactivator Assay in antagonist mode (to identify antagonist compounds), PR-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluorescein-labeled coactivator peptide, and terbium-labeled anti-GST antibody (see figure). The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

PolarScreen™ Androgen Receptor Competitor Assay Kit, Green

The PolarScreen™ Androgen Receptor Competitor Assay Kit, Green, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential androgen receptor (AR) ligands. The kit uses insect cell-expressed, rat AR-ligand binding domain tagged with glutathione-S-transferase (GST) and histidine (also available separately), plus a novel, high-affinity, fluorescent androgen ligand (Fluormone™ AL Green) in a homogenous, mix-and-read assay format.

This kit produces a green color read-out; a "red" 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 us at drugdiscoverytech@lifetech.com.

The PolarScreen Androgen Receptor Competitor Assay is:

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

How the Assay Works
The kit uses the rat AR ligand-binding domain tagged with His and GST [AR-LBD(His-GST)]. AR-LBD(His-GST) is added to a fluorescently-tagged androgen ligand (FluormoneAL Green) in the presence of competitor test compounds in microwell plates. The presence of effective competitors prevents the formation of a AL Green/AR-LBD(His-GST) complex, resulting in a decrease of the polarization value due to ligand displacement caused by a competitor. The shift in polarization value in the presence of test compounds is used to determine relative affinity of test compounds for AR-LBD(His-GST).

Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

Hemoglobin Colorimetric Detection Kit (Invitrogen™)

The Hemoglobin Detection research-use-only kit is a colorimetric assay designed for the quantification and detection of hemoglobin in Whole Blood, RBCs, and hemolyzed serum and plasma samples.

This complete, ready-to-use kit includes clear 96-well plate(s), hemoglobin standard, and other components to perform the assay. A 96-well microplate reader capable of reading optical density between 560 and 580 nm is required for use of this kit.

Performance characteristics
• Assay type: colorimetric detection kit
• Sample types: whole blood, RBCs, and hemolyzed serum and plasma samples
• Sensitivity 0.021 g/dL
• Standard curve range: 0.25 g/dL–16 g/dL
• Reactivity: species independent

Background
Hemoglobin (Hgb) is an erythrocyte protein complex comprised of two sets of identical pairs of subunits, each of which bind an iron-prophyrin group commonly called heme. Generally containing two alpha or alpha-like globulin chains, the remaining subunits may be beta, gamma, delta or epsilon, or in the case of infants, fetal hemoglobin that is replaced during the first year of life. Heme binds and releases oxygen or carbon dioxide in response to slight changes in local gas tension. Free oxygen or carbon dioxide bound by one heme group facilitates subsequent binding by the other heme groups in a given hemoglobin molecule.

Assay principle
The Hemoglobin Colorimetric Detection Kit is designed to quantitatively measure all forms of hemoglobin present in blood and RBCs, or plasma and serum. A human hemoglobin standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. Standards or diluted samples are pipetted into a clear microtiter plate and the ready-to-use Hemoglobin Detection Reagent is added to each well. For whole blood or RBC samples, 10 μL of samples and standards are used in the Regular format. Results are calculated as g/dL for whole blood and RBCs, and mg/mL for serum and plasma. The plate is incubated for 30 minutes at room temperature. The plate is read at 560-580 nm to detect the intensity of the color generated. This Hemoglobin Detection kit uses a single reaction solution that is light stable at 4°C and does not contain dangerous chemicals. All forms of hemoglobin are rapidly converted to a single stable form that is measured photometrically. Many samples can be measured without dilution in this safe, simple assay.

Related links
Learn more about ELISA kits
Learn more about other immunoassays

LanthaScreen™ TR-FRET ERR beta Coactivator Assay Kit, goat

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15139:

The LanthaScreen® TR-FRET ERR (estrogen related receptor) beta Coactivator Assay Kit provides a sensitive and robust method for highthroughput screening of potential ERR beta ligands as inverse agonists of ligand-dependent coactivator displacement. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant ERR beta ligand-binding domain (ERR beta-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.

To assay:

When running the LanthaScreen® ERR beta TR-FRET Coactivator Assay, ERR beta-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 ERR 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 displace coactivator peptide (see Figure 1).

PolarScreen™ Progesterone Receptor Competitor Assay Kit, Green

The PolarScreen™ Progesterone Receptor Competitor Assay Kit, Green, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential progesterone receptor (PR) ligands. The kit uses insect cell-expressed, human PR ligand binding domain tagged with glutathione-S-transferase (GST) (also available separately) and a novel, high-affinity, fluorescent progesterone receptor ligand (Fluormone™ PL Green) in a homogenous, mix-and-read assay format.

This kit produces a green color read-out; a "red" 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 us at drugdiscoverytech@lifetech.com.

The PolarScreen Progesterone Receptor Competitor Assay is:

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

How the Assay Works
An N-terminal fusion of glutathione transferase to the ligand-binding domain of the human progesterone receptor (PR-LBD(GST)) is added to a fluorescently-tagged progesterone ligand (Fluormone PL Green) in the presence of competitor test compounds in microwell plates. The presence of effective competitors prevents the formation of a PL Green/PR-LBD(GST) complex, resulting in a decrease of the polarization value. The shift in polarization value in the presence of test compounds is used to determine relative affinity of test compounds for PR-LBD(GST).

Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

PolarScreen™ Androgen Receptor Competitor Assay Kit, Red

The PolarScreen™ Androgen Receptor Competitor Assay Kit, Red, provides a sensitive and efficient method for high-throughput, fluorescence polarization-based screening of potential androgen receptor (AR) ligands. The kit uses insect cell-expressed rat AR ligand binding domain tagged with glutathione-S-transferase (GST) and histidine (also available separately), plus a novel, high-affinity, fluorescent androgen receptor ligand (Fluormone AL 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 us at drugdiscoverytech@lifetech.com.

The PolarScreen Androgen Receptor Competitor Assay is:

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

How the Assay Works
The kit uses the rat AR ligand-binding domain tagged with His and GST [AR-LBD(His-GST)]. AR-LBD(His-GST) is added to a fluorescently-tagged androgen ligand (FluormoneAL Red) in the presence of competitor test compounds in microwell plates. Competitors displace the fluorescent FluormoneAL Red Ligand from the AR-LBD/FluormoneAL Red complex, causing the fluorescent ligand to tumble rapidly during its fluorescence lifetime, resulting in a low polarization value. Noncompetitors will not displace the fluorescent ligand from the complex; therefore, the polarization value remains high. The shift in polarization value in the presence of the test compounds is used to determine relative affinity of test compounds for the AR-LBD.

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 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 gamma Coactivator Assay Kit, rabbit

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4548:

The LanthaScreen® TR-FRET PPAR gamma Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential PPAR gamma 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 Peroxisome Proliferator Activated Receptor (PPAR) gamma ligand-binding domain (PPAR gamma-LBD) that is tagged with glutathione-S-transferase (GST), in a homogenous mix-and-read assay format.

Agonist mode
To run the LanthaScreen® TR-FRET Peroxisome Proliferator Activated gamma Receptor Coactivator Assay in agonist mode (to identify agonist compounds), PPAR gamma-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti-GST antibody. After an incubation period at room temperature, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR gamma-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).

Antagonist mode
When the LanthaScreen® TR-FRET Peroxisome Proliferator Activated gamma Receptor Coactivator Assay is run in antagonist mode (to identify antagonist compounds), PPAR gamma-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluores-cein-coactivator peptide, and Tb-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 (Figure 2).

Contents and Storage:
The LanthaScreen® TR-FRET PPARγ Coactivator Assay Kit contains PPAR gamma-LBD (GST) protein, fluorescently 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.

PolarScreen™ PPARγ-Competitor Assay Kit, Green

The PPARγ (Peroxisome Proliferator-Activated Receptor-gamma) Competitor Assay, Green uses the human-derived recombinant PPARγ ligand-binding domain (PPARγ-LBD) tagged with an N-terminal GST-tag and a novel, tight-binding, selective fluorescent PPARγ ligand (Fluormone™ PPARγ Green) in a homogenous, mix-and-read assay format. PPARγ-LBD/Fluormone™ PPARγ Green complex produces high polarization. This complex is then added to individual test compounds in microwell plates. Competitors displace the fluorescent Fluormone™ PPARγ Green Ligand from the PPARγ-LBD/Fluormone™ PPARγ Green complex, causing the fluorescent ligand to tumble rapidly during its fluorescence lifetime, resulting in a low polarization value. Noncompetitors will not displace the fluorescent ligand from the complex, so the polarization value remains high. The shift in polarization value in the presence of the test compounds is used to determine relative affinity of test compounds for the PPARγ-LBD.

PiPer™ Pyrophosphate Assay Kit (Invitrogen™)

The PiPer™ Pyrophosphate Assay Kit provides a sensitive fluorogenic or chromogenic method for detecting as little as 0.4 µM (fluorescence) or 2 µM (absorbance) inorganic pyrophosphate in a 100 µL assay volume.