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Human PEBP4(Phosphatidylethanolamine Binding Protein 4) ELISA Kit (HUES03439)

SKU:
HUES03439
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q96S96
Sensitivity:
0.09ng/mL
Range:
0.16-10ng/mL
ELISA Type:
Sandwich
Reactivity:
Human
Sample Type:
Serum, plasma and other biological fluids
Research Area:
Signal Transduction
€599
Frequently bought together:

Description

system_update_altDatasheetsystem_update_altMSDS
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 PEBP4 in samples. No significant cross-reactivity or interference between Human PEBP4 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 PEBP4. 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 PEBP4 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 PEBP4, 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 PEBP4. The concentration of Human PEBP4 in samples can be calculated by comparing the OD of the samples to the standard curve.

UniProt Protein Function:PEBP4: Seems to promote cellular resistance to TNF-induced apoptosis by inhibiting activation of the Raf-1/MEK/ERK pathway, JNK and phosphatidylethanolamine externalization. Belongs to the phosphatidylethanolamine-binding protein family.
UniProt Protein Details:

Protein type:Secreted, signal peptide; Lipid-binding; Secreted

Chromosomal Location of Human Ortholog: 8p21. 3

Molecular Function:protein binding

NCBI Summary:The phosphatidylethanolamine (PE)-binding proteins, including PEBP4, are an evolutionarily conserved family of proteins with pivotal biologic functions, such as lipid binding and inhibition of serine proteases (Wang et al. , 2004 [PubMed 15302887]). [supplied by OMIM, Dec 2008]
UniProt Code:Q96S96
NCBI GenInfo Identifier:143811436
NCBI Gene ID:157310
NCBI Accession:Q96S96. 3
UniProt Secondary Accession:Q96S96,Q5EVA1, Q8WW74,
UniProt Related Accession:Q96S96
Molecular Weight:25,733 Da
NCBI Full Name:Phosphatidylethanolamine-binding protein 4
NCBI Synonym Full Names:phosphatidylethanolamine binding protein 4
NCBI Official Symbol:PEBP4
NCBI Official Synonym Symbols:CORK1; CORK-1; PEBP-4; hPEBP4; PRO4408; GWTM1933; HEL-S-300
NCBI Protein Information:phosphatidylethanolamine-binding protein 4
UniProt Protein Name:Phosphatidylethanolamine-binding protein 4
UniProt Synonym Protein Names:Protein cousin-of-RKIP 1
Protein Family:Phosphatidylethanolamine-binding protein
UniProt Gene Name:PEBP4
UniProt Entry Name:PEBP4_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
102.57
2.59		2.582.493
51.596
1.636		1.6161.529
2.50.997
0.963		0.980.893
1.250.473
0.497		0.4850.398
0.630.266
0.264		0.2650.178
0.320.203
0.181		0.1920.105
0.160.138
0.144		0.1410.054
00.083
0.091		0.087--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PEBP4 were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PEBP4 were tested on 3 different plates, 20 replicates in each plate.

Intra-assay PrecisionInter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)0.460.833.550.420.833.80
Standard deviation0.030.040.150.030.030.13
C V (%)6.524.824.237.143.613.42

Recovery

The recovery of Human PEBP4 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)83-9690
EDTA plasma (n=5)95-107101
Cell culture media (n=5)85-9992

Linearity

Samples were spiked with high concentrations of Human PEBP4 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 (%)88-10093-10690-101
Average (%)949996
1:4Range (%)90-10486-10182-93
Average (%)969287
1:8Range (%)91-10781-9381-93
Average (%)988787
1:16Range (%)89-10184-9488-99
Average (%)948994

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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