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PLTP Inhibitor Drug Screening Kit (Fluorometric)

Catalog #: K605

In stock

$520.00

Product Details

Cat # +Size K605-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 465/535 nm)
Species Reactivity Mammalian
Applications N/A
Features & Benefits • Simple procedure; takes ~40 minutes
• Fast and convenient
• Provided all necessary buffers and reagents, as well as positive control for accurate assay of activity of plasma phospholipid transfer protein (PLTP).
Kit Components • PLTP Donor Molecule
• PLTP Acceptor Molecule
• PLTP Assay Buffer (10X)
• Positive Control (Rabbit Serum)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Plasma phospholipid transfer protein (PLTP) is thought to play a major role in the facilitated transfer of phospholipids between lipoproteins and in the modulation of high-density lipoprotein (HDL) particle size and composition. PLTP-facilitated lipid transfer activity is related to HDL and LDL metabolism, as well as lipoprotein lipase activity, adiposity, and insulin resistance. The PLTP Drug Screening Kit utilizes rabbit serum as PLTP activity source to screen PLTP inhibitors. The assay uses a donor molecule containing a fluorescent self-quenched phospholipid that is transferred to an acceptor molecule in the presence of PLTP. PLTP-mediated transfer of the fluorescent phospholipid to the acceptor molecule results in an increase in fluorescence (Ex/ Em- 465/ 535 nm). Inhibitor of PLTP will inhibit the lipid transfer and therefore decrease fluorescence intensity. The kit provides sufficient reagents for 100 PLTP inhibitor screening assays.


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Does this kit work with human or mouse samples?
We use rabbit serum as our positive control in this assay. This assay should work with humans or mice too. The main point is to have active PLTP enzyme in your sample.
How can sevarl inhibitors be compared using this assay?
To compare several inhibitors, a rank ordering system can be used based on relative inhibitory potential with respect to the positive control. Different concentrations/amounts of sample can be tested to generate IC50 curves.
The value from the blank is very high, higher than the sample. What should I do?
This is where kinetics becomes important. The blank RFU values will be stable over time. The solution to this problem is to select a timepoint where the sample values are increasing linearly and within the range of the standard curve, while being higher than the blank reading. The increase in fluorescenceintensity is usually 0.2 2 foldover blank. It is important to optimize the sample volume to be used for the assay, since saturating amounts of sample can lead to difficulty in selecting the timepoint with a linear increase in enzyme activity.
After incubating for 2 hours, the sample reading is lower than the reading at 10 mins. Which time points should I use for the calculations?
To select the correct time points, the entire kinetic curve needs to be measured. Select two time points within which the increase in RFU is linear and within the std. curve range. Sealing the plate is critical to prevent evaporation and to avoid concentrating the sample during incubation at 37C. Using values without the plate seal can lead to overestimation of PLTP activity in the sample.
Can frozen tissue be used with this kit?
Although not ideal, flash-frozen tissue or cells stored at -80C can be used. It is important to remember that results may vary between fresh and frozen samples.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Are trial sizes of this kit available?
Unfortunately, we do not have trial sizes of this kit available. However,for customers based in the US or Canada, a 10% off list price introductory discount can be offered on the list price. For all other customers based out of this area our regional BioVision distributor should be contacted"
How do I normalize my samples against protein concentration
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample diluted in the assay buffer.
Can we buy individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffer is optimized for the conditions required by the PLTP enzyme to work optimally. Therefore, we highly recommend using the buffer provided in the kit for the best results.