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Human PPAR-delta (Peroxisome Proliferator Activated Receptor Delta) ELISA Kit (HUES03266)

SKU:
HUES03266
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q03181
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Epigenetics and Nuclear Signaling
$719
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Description

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Human PPAR-delta (Peroxisome Proliferator Activated Receptor Delta) ELISA Kit

The Human PPAR Delta (Peroxisome Proliferator-Activated Receptor Delta) ELISA Kit is a valuable tool for accurately measuring PPAR Delta levels in human samples such as serum, plasma, and cell culture supernatants. This kit provides a high level of sensitivity and specificity, ensuring precise and consistent results for various research applications.PPAR Delta is a key nuclear receptor involved in regulating lipid metabolism, inflammation, and energy homeostasis.

Dysregulation of PPAR Delta has been linked to conditions such as obesity, diabetes, and cardiovascular diseases, making it a valuable biomarker for studying these diseases and potential therapeutic interventions. By using the Human PPAR Delta ELISA Kit, researchers can gain valuable insights into the role of PPAR Delta in health and disease, paving the way for new discoveries and advancements in the field of metabolic disorders and related conditions.

Assay type:Sandwich
Format:96T
Assay time:4.5h
Reactivity:Human
Detection Method:Colormetric
Detection Range:0.16-10 ng/mL
Sensitivity:0.10 ng/mL
Sample Volume Required Per Well:100µL
Sample Type:Serum, plasma and other biological fluids
Specificity:This kit recognizes Human PPAR-delta in samples. No significant cross-reactivity or interference between Human PPAR-delta and analogues was observed.

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human PPAR-delta. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human PPAR-delta and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human PPAR-delta, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human PPAR-delta. The concentration of Human PPAR-delta in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:Ligand-activated transcription factor. Receptor that binds peroxisome proliferators such as hypolipidemic drugs and fatty acids. Has a preference for poly-unsaturated fatty acids, such as gamma-linoleic acid and eicosapentanoic acid. Once activated by a ligand, the receptor binds to promoter elements of target genes. Regulates the peroxisomal beta-oxidation pathway of fatty acids. Functions as transcription activator for the acyl-CoA oxidase gene. Decreases expression of NPC1L1 once activated by a ligand.
NCBI Summary:This gene encodes a member of the peroxisome proliferator-activated receptor (PPAR) family. PPARs are nuclear hormone receptors that bind peroxisome proliferators and control the size and number of peroxisomes produced by cells. PPARs mediate a variety of biological processes, and may be involved in the development of several chronic diseases, including diabetes, obesity, atherosclerosis, and cancer. This protein is a potent inhibitor of ligand-induced transcription activity of PPAR alpha and PPAR gamma. It may function as an integrator of transcription repression and nuclear receptor signaling. The expression of this gene is found to be elevated in colorectal cancer cells. The elevated expression can be repressed by adenomatosis polyposis coli (APC), a tumor suppressor protein related to APC/beta-catenin signaling pathway. Knockout studies in mice suggested the role of this protein in myelination of the corpus callosum, lipid metabolism, and epidermal cell proliferation. Alternate splicing results in multiple transcript variants. [provided by RefSeq, Jan 2010]
UniProt Code:Q03181
NCBI GenInfo Identifier:417522
NCBI Gene ID:5467
NCBI Accession:Q03181. 1
UniProt Secondary Accession:Q03181,Q5D1P0, Q7Z5K0, Q9BUD4, A8K6J6, B4E3V3, B6ZGS1 B7Z3W1, E9PE18,
UniProt Related Accession:Q03181
Molecular Weight:38,855 Da
NCBI Full Name:Peroxisome proliferator-activated receptor delta
NCBI Synonym Full Names:peroxisome proliferator activated receptor delta
NCBI Official Symbol:PPARD
NCBI Official Synonym Symbols:FAAR; NUC1; NUCI; NR1C2; NUCII; PPARB
NCBI Protein Information:peroxisome proliferator-activated receptor delta
UniProt Protein Name:Peroxisome proliferator-activated receptor delta
UniProt Synonym Protein Names:NUCI; Nuclear hormone receptor 1; NUC1; Nuclear receptor subfamily 1 group C member 2; Peroxisome proliferator-activated receptor beta; PPAR-beta
Protein Family:Peroxisome proliferator-activated receptor
UniProt Gene Name:PPARD
UniProt Entry Name:PPARD_HUMAN

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e. g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(ng/mL)		O.DAverageCorrected
102.533
2.569		2.5512.481
51.672
1.672		1.6721.602
2.50.985
0.957		0.9710.901
1.250.518
0.53		0.5240.454
0.630.263
0.245		0.2540.184
0.320.181
0.169		0.1750.105
0.160.124
0.124		0.1240.054
00.064
0.076		0.07--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PPAR-delta were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PPAR-delta were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.490.904.810.520.954.67
Standard deviation0.030.050.230.030.050.21
C V (%)6.125.564.785.775.264.50

Recovery

The recovery of Human PPAR-delta spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample TypeRange (%)Average Recovery (%)
Serum (n=5)93-10698
EDTA plasma (n=5)86-9892
Cell culture media (n=5)90-10596

Linearity

Samples were spiked with high concentrations of Human PPAR-delta and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5)EDTA plasma (n=5)Cell culture media (n=5)
1:2Range (%)84-9985-9695-109
Average (%)9191103
1:4Range (%)94-10882-9582-95
Average (%)998989
1:8Range (%)94-10586-9988-99
Average (%)999293
1:16Range (%)89-10181-9185-96
Average (%)958690

An unopened kit can be stored at 4°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
Reference Standard 2 vials
Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 µL
Concentrated HRP Conjugate (100×) 1 vial, 120 µL -20°C(shading light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 4°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 4°C(shading light)
Stop Solution 1 vial, 10 mL 4°C
Plate Sealer 5 pieces
Product Description 1 copy
Certificate of Analysis 1 copy
  1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
  2. Aliquot 100µl of standard solutions into the standard wells.
  3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
  4. Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
  5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
  6. Aspirate the liquid from each well, do not wash. Immediately add 100µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix. Incubate for 1 hour at 37°C.
  7. Aspirate or decant the solution from the plate and add 350µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
  8. Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
  9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
  10. Add 90µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37°C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
  11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
  12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

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