AGILENT SEAHORSE XF PALMITATE-BSA FAO SUBSTRATE · Agilent Products are for Research Use Only. Not...
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AGILENT SEAHORSE XF PALMITATE-BSA FAO SUBSTRATE A Substrate for Measuring Long Chain Fatty Acid Oxidation (FAO) in the Agilent Seahorse XF Analyzer Agilent Seahorse XF Palmitate-BSA FAO Substrate • Ready-to-use solution for consistent results in every assay. Free fatty acid concentration (FFA) listed with each lot • Reliable, functional information without radioactivity • Improved workflow and integration with the Agilent Seahorse XF Cell Mito Stress Test For more information, visit: www.agilent.com Measure Fatty Acid Oxidation (FAO) in Cells, in Minutes Agilent Seahorse XF Palmitate-BSA FAO Substrate is a preconjugated, quality-controlled, fatty acid substrate that simplifies the process for obtaining functional data to characterize the metabolic phenotype of your cells. This bioavailable, conjugated fatty acid substrate frees you from the tedious, time-consuming, and variable conjugation process. Agilent Seahorse XF Palmitate-BSA FAO Substrate Using the Seahorse XF Palmitate-BSA FAO Substrate, researchers can now determine precisely how a drug of interest, treatment, or genetic modification, changes the ability of a cell to oxidize fatty acids. Many cells store triglycerides and other energy stores that can affect the rates of respiration when fatty acids are added to the experimental media. The Seahorse XF Palmitate-BSA FAO Substrate can be used with the Agilent Seahorse XF Cell Mito Stress Test to quantify oxidation of both endogenous and exogenously provided long chain fatty acids. Figure 1. Integration of the Agilent Seahorse XF Palmitate-BSA FAO Substrate with the Agilent Seahorse XF Cell Mito Stress Test. Distinguish uncoupling from actual fatty acid oxidation. The oligomycin injection provides information regarding mild uncoupling by free fatty acids. Increases in maximal respiration beyond the BSA control indicate utilization of exogenous FFAs. 0 10 20 30 40 50 0 20 40 60 80 100 120 140 160 Time (min) Oxidation of exogenous FAs + uncoupling by FAs (D15 pmol/min O 2 ) Uncoupling by FAs (D6 pmol/min O 2 ) OCR (pmol/min) 1 2 3 1. 2. 3.Oxidation of exogenous FAs (D87 pmol/min O 2 ) BSA Palm: BSA
Transcript of AGILENT SEAHORSE XF PALMITATE-BSA FAO SUBSTRATE · Agilent Products are for Research Use Only. Not...
AGILENT SEAHORSE XFPALMITATE-BSA FAO SUBSTRATE
A Substrate for Measuring Long Chain Fatty Acid Oxidation (FAO) in the Agilent Seahorse XF Analyzer
Agilent Seahorse XF Palmitate-BSA FAO Substrate
• Ready-to-use solution for consistent results in every assay. Free fatty acid concentration (FFA) listed with each lot
• Reliable, functional information without radioactivity
• Improved workflow and integration with the Agilent Seahorse XF Cell Mito Stress Test
For more information, visit:www.agilent.com
Measure Fatty Acid Oxidation (FAO) in Cells, in Minutes
Agilent Seahorse XF Palmitate-BSA FAO Substrate is a preconjugated, quality-controlled, fatty acid substrate that simplifies the process for obtaining functional data to characterize the metabolic phenotype of your cells. This bioavailable, conjugated fatty acid substrate frees you from the tedious, time-consuming, and variable conjugation process.
Agilent Seahorse XF Palmitate-BSA FAO Substrate
Using the Seahorse XF Palmitate-BSA FAO Substrate, researchers can now determine precisely how a drug of interest, treatment, or genetic modification, changes the ability of a cell to oxidize fatty acids. Many cells store triglycerides and other energy stores that can affect the rates of respiration when fatty acids are added to the experimental media. The Seahorse XF Palmitate-BSA FAO Substrate can be used with the Agilent Seahorse XF Cell Mito Stress Test to quantify oxidation of both endogenous and exogenously provided long chain fatty acids.
Figure 1. Integration of the Agilent Seahorse XF Palmitate-BSA FAO Substrate with the Agilent Seahorse XF Cell Mito Stress Test. Distinguish uncoupling from actual fatty acid oxidation. The oligomycin injection provides information regarding mild uncoupling by free fatty acids. Increases in maximal respiration beyond the BSA control indicate utilization of exogenous FFAs.
0 10 20 30 40 500
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160
Time (min)
Oxidation of exogenousFAs + uncoupling by FAs(D15 pmol/min O2)Uncoupling by FAs(D6 pmol/min O2)
OCR
(pm
ol/m
in)
12
3
1.
2.
3.Oxidation of exogenous FAs(D87 pmol/min O2)
BSAPalm: BSA
Agilent Products are for Research Use Only.Not for use in diagnostic procedures.
Information, descriptions and specifications in thispublication are subject to change without notice.
© Agilent Technologies, Inc. 2016Published in USA, December 1, 2016
5991-7122EN
For more information, visit: www.agilent.com
Determination of Exogenous Fatty Acid Utilization in the Agilent Seahorse XF Analyzer
In Figure 2 and Figure 3, the utilization of exogenous fatty acids is dependent upon placing energetic stress (via FCCP) on the cells. This experiment illustrates that under conditions of substrate limitation, and the combined use of the Seahorse XF Palmitate-BSA FAO Substrate, etomoxir (Eto) and the Seahorse XF Cell Mito Stress Test, one can determine the proportion of respiration that is supported by exogenous fatty acids.
Figure 2. A kinetic graph illustrating the use of the Agilent Seahorse XF Palmitate-BSA Substrate, followed by the Agilent Seahorse XF Cell Mito Stress Test. The red arrow indicates exogenous fatty acid utilization.
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Time (min)
Oligomycin FCCP Rtn/AA
OCR
(pm
ol/m
in)
BSA, –EtoBSA, +Eto
1. Oxidation of exogenous palmitate
Palm: BSA, –EtoPalm: BSA, +Eto
1
Figure 3. Bar chart highlighting the differences in maximal respiration among groups. The data is taken from rate measurement "10" in Figure 2.
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OCR
(pm
ol/m
in)
BSA, –Eto
BSA, +Eto
Palm: B
SA, –Eto
Palm: B
SA, +Eto
BSA, –EtoBSA, +EtoPalm: BSA, –EtoPalm: BSA, +Eto
Component DescriptionProduct Agilent Seahorse XF Palmitate-BSA FAO SubstratePart number 102720-100Kit contents Agilent Seahorse XF Palmitate-BSA: 1 mM solution
(3 × 2 mL vials) Agilent Seahorse XF BSA Control: 0.17 mM solution (3 × 2 mL vials)
Shipping conditions Ships at ambient temperature, packed in insulated storage box
Storage requirements Store at –20 °CNote: One vial of XF Palmitate-BSA or XF BSA control contains enough
material for one Agilent Seahorse XF96/XF24 Microplate.
Product Information
