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Citations & References (13)
Invitrogen™
Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit
The Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit provides a sensitive method for detecting phosphatidylcholine-specific phospholipase C (PC-PLC) activity usingRead more
| Catalog Number | Quantity |
|---|---|
| A12218 | 500 Assays |
Catalog number A12218
Price (CNY)
6,036.00
Each
Quantity:
500 Assays
Price (CNY)
6,036.00
Each
The Amplex™ Red Phosphatidylcholine-Specific Phospholipase C Assay Kit provides a sensitive method for detecting phosphatidylcholine-specific phospholipase C (PC-PLC) activity using a fluorescence microplate reader or fluorometer.
See our complete line of Fluorescence Microplate assays.
• Detects phospholipase-C activity levels as low as 0.2 mU/mL
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference
In this enzyme-coupled assay, PC-PLC activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex™ Red reagent), a sensitive fluorogenic probe for hydrogen peroxide. PC-PLC converts phosphatidylcholine (lecithin) to phosphocholine and diacylglycerol. Alkaline phosphatase is added, which hydrolyzes phosphorylcholine to form choline. Choline is then oxidized by choline oxidase to form betaine and hydrogen peroxide. In the presence of horseradish peroxidase, the hydrogen peroxide reacts with Amplex™ Red reagent in a 1:1 stoichiometric ratio to generate the highly fluorescent product resorufin.
Because resorufin has absorption and fluorescence emission maxima of approximately 571 nm and 585 nm, respectively, there is little interference from autofluorescence in most biological samples.
Use Amplex™ Red Assays for a Broad Range of Investigations
A wide variety of validated Amplex™ Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex™ UltraRed Reagent (Cat. No. A36006), a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex™ Red/UltraRed Stop Reagent (Cat. No. A33855). The Amplex™ Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least three hours. Custom assay design and packaging are also available.
See our complete line of Fluorescence Microplate assays.
• Detects phospholipase-C activity levels as low as 0.2 mU/mL
• Format allows for multiple time point measurements
• Designed for minimal autofluorescence interference
In this enzyme-coupled assay, PC-PLC activity is monitored indirectly using 10-acetyl-3,7-dihydroxyphenoxazine (Amplex™ Red reagent), a sensitive fluorogenic probe for hydrogen peroxide. PC-PLC converts phosphatidylcholine (lecithin) to phosphocholine and diacylglycerol. Alkaline phosphatase is added, which hydrolyzes phosphorylcholine to form choline. Choline is then oxidized by choline oxidase to form betaine and hydrogen peroxide. In the presence of horseradish peroxidase, the hydrogen peroxide reacts with Amplex™ Red reagent in a 1:1 stoichiometric ratio to generate the highly fluorescent product resorufin.
Because resorufin has absorption and fluorescence emission maxima of approximately 571 nm and 585 nm, respectively, there is little interference from autofluorescence in most biological samples.
Use Amplex™ Red Assays for a Broad Range of Investigations
A wide variety of validated Amplex™ Red assays are available for studying cell signaling and lipids, neurobiology, inflammation and immune function, and metabolism. We also offer Amplex™ UltraRed Reagent (Cat. No. A36006), a second-generation reagent providing greater sensitivity and brighter fluorescence, and the Amplex™ Red/UltraRed Stop Reagent (Cat. No. A33855). The Amplex™ Red/UltraRed Stop Reagent provides convenience and control by allowing the fluorescence signal-generating reaction to be terminated at a user-determined time point. After addition of the stop reagent, the fluorescence signal remains stable for at least three hours. Custom assay design and packaging are also available.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodFluorescence
Dye TypeOther Label(s) or Dye(s)
FormatCuvettes, 96-well plate
Quantity500 Assays
Shipping ConditionRoom Temperature
For Use With (Application)Phospholipase Assay
For Use With (Equipment)Fluorometer, Microplate Reader
Product LineAmplex
Product TypeAmplex Red Assay Kit
Unit SizeEach
Contents & Storage
Store in freezer -5°C to -30°C and protect from light.
Frequently asked questions (FAQs)
I'm using an Amplex Red kit, the reagent changes color to pink almost immediately in my own Krebs-Ringer buffer but not in HBSS. Why is this?
Can Amplex Red Assays be performed using cell lysates?
Citations & References (13)
Citations & References
Abstract
CDP-choline significantly restores phosphatidylcholine levels by differentially affecting phospholipase A2 and CTP: phosphocholine cytidylyltransferase after stroke.
Journal:J Biol Chem
PubMed ID:16380371
'Phosphatidylcholine (PtdCho) is a major membrane phospholipid, and its loss is sufficient in itself to induce cell death. PtdCho homeostasis is regulated by the balance between hydrolysis and synthesis. PtdCho is hydrolyzed by phospholipase A2 (PLA2), PtdChospecific phospholipase C (PtdCho-PLC), and phospholipase D (PLD). PtdCho synthesis is rate-limited by CTP:phosphocholine
The antisignaling agent SC-alpha alpha delta 9, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid, is a structurally unique phospholipid analogue with phospholipase C inhibitory activity.
Journal:Mol Cancer Ther
PubMed ID:12481409
'Phospholipids and lipid second messengers mediate mitogenic signal transduction and oncogenesis, but there have been few successful examples of small molecules that affect biologically important phospholipid metabolism. Here we investigated the actions of a previously described antitumor agent, 4-(benzyl-(2-[(2,5-diphenyloxazole-4-carbonyl)amino]ethyl)carbamoyl)- 2-decanoylaminobutyric acid (SC-alpha alpha delta 9), which has antisignaling properties, on
Defective coupling of apical PTH receptors to phospholipase C prevents internalization of the Na+-phosphate cotransporter NaPi-IIa in Nherf1-deficient mice.
Journal:Am J Physiol Cell Physiol
PubMed ID:16987995
'Phosphate reabsorption in the renal proximal tubule occurs mostly via the type IIa Na(+)-phosphate cotransporter (NaP(i)-IIa) in the brush border membrane (BBM). The activity and localization of NaP(i)-IIa are regulated, among other factors, by parathyroid hormone (PTH). NaP(i)-IIa interacts in vitro via its last three COOH-terminal amino acids with the
Enzymatic measurement of phosphatidylserine in cultured cells.
Journal:J Lipid Res
PubMed ID:22100437
Phosphatidylserine (PS) is a quantitatively minor membrane phospholipid involved in diverse cellular functions. In this study, we developed a new fluorometric method for measuring PS using combinations of specific enzymes and Amplex Red. The calibration curve for PS measurement was linear and hyperbolic at low (0-50 µM) and high (50-1000
Phosphatidylcholine-specific phospholipase C activation is required for CCR5-dependent, NF-kB-driven CCL2 secretion elicited in response to HIV-1 gp120 in human primary macrophages.
Journal:Blood
PubMed ID:18203956
CCL2 (MCP-1) has been shown to enhance HIV-1 replication. The expression of this chemokine by macrophages is up-modulated as a consequence of viral infection or gp120 exposure. In this study, we show for the first time that the phosphatidylcholine-specific phospholipase C (PC-PLC) is required for the production of CCL2 triggered