Human Cardiovascular ELISA Kits
Human PGF (Placental Growth Factor) ELISA Kit (HUES02569)
- SKU:
- HUES02569
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- P49763
- Sensitivity:
- 9.38pg/mL
- Range:
- 15.63-1000pg/mL
- ELISA Type:
- Sandwich
- Synonyms:
- PLGF, PGFL, PlGF-2, SHGC-10760, D12S1900
- Reactivity:
- Human
- Sample Type:
- Serum, plasma and other biological fluids
- Research Area:
- Cardiovascular
Description
| Assay type: | Sandwich |
| Format: | 96T |
| Assay time: | 4.5h |
| Reactivity: | Human |
| Detection Method: | Colormetric |
| Detection Range: | 15.63-1000 pg/mL |
| Sensitivity: | 9.38 pg/mL |
| Sample Volume Required Per Well: | 100µL |
| Sample Type: | Serum, plasma and other biological fluids |
| Specificity: | This kit recognizes Human PGF in samples. No significant cross-reactivity or interference between Human PGF 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 PGF. 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 PGF 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 PGF, 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 PGF. The concentration of Human PGF in samples can be calculated by comparing the OD of the samples to the standard curve.
| UniProt Protein Function: | PGF: Growth factor active in angiogenesis and endothelial cell growth, stimulating their proliferation and migration. It binds to the receptor FLT1/VEGFR-1. Isoform PlGF-2 binds NRP1/neuropilin-1 and NRP2/neuropilin-2 in a heparin-dependent manner. Also promotes cell tumor growth. Belongs to the PDGF/VEGF growth factor family. 3 isoforms of the human protein are produced by alternative splicing. |
| UniProt Protein Details: | Protein type:Secreted; Secreted, signal peptide Chromosomal Location of Human Ortholog: 14q24. 3 Cellular Component: extracellular space; membrane; extracellular region Molecular Function:heparin binding; protein binding; protein homodimerization activity; growth factor activity; protein heterodimerization activity Biological Process: response to drug; organ regeneration; female pregnancy; signal transduction; cellular response to hormone stimulus; positive regulation of angiogenesis; cell-cell signaling; ureteric bud branching; positive regulation of cell division; positive regulation of cell proliferation; response to hypoxia; positive regulation of endothelial cell proliferation; sprouting angiogenesis; vascular endothelial growth factor receptor signaling pathway; cell differentiation |
| NCBI Summary: | This gene encodes a growth factor found in placenta which is homologous to vascular endothelial growth factor. Alternatively spliced transcripts encoding different isoforms have been found for this gene. [provided by RefSeq, Jun 2011] |
| UniProt Code: | P49763 |
| NCBI GenInfo Identifier: | 17380553 |
| NCBI Gene ID: | 5228 |
| NCBI Accession: | P49763. 2 |
| UniProt Secondary Accession: | P49763,Q07101, Q9BV78, Q9Y6S8, |
| UniProt Related Accession: | P49763 |
| Molecular Weight: | 19,338 Da |
| NCBI Full Name: | Placenta growth factor |
| NCBI Synonym Full Names: | placental growth factor |
| NCBI Official Symbol: | PGF |
| NCBI Official Synonym Symbols: | PGFL; PLGF; PlGF-2; D12S1900; SHGC-10760 |
| NCBI Protein Information: | placenta growth factor; placental growth factor, vascular endothelial growth factor-related protein |
| UniProt Protein Name: | Placenta growth factor |
| Protein Family: | Placenta growth factor |
| UniProt Gene Name: | PGF |
| UniProt Entry Name: | PLGF_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 (pg/mL) | O.D | Average | Corrected |
| 1000 | 2.522 2.554 | 2.538 | 2.46 |
| 500 | 1.568 1.614 | 1.591 | 1.513 |
| 250 | 0.919 0.897 | 0.908 | 0.83 |
| 125 | 0.468 0.49 | 0.479 | 0.401 |
| 62.5 | 0.265 0.247 | 0.256 | 0.178 |
| 31.25 | 0.18 0.178 | 0.179 | 0.101 |
| 15.63 | 0.121 0.139 | 0.13 | 0.052 |
| 0 | 0.076 0.08 | 0.078 | -- |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human PGF were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human PGF were tested on 3 different plates, 20 replicates in each plate.
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 47.69 | 110.91 | 352.01 | 48.61 | 111.35 | 363.77 |
| Standard deviation | 2.39 | 5.69 | 11.76 | 2.55 | 5.45 | 12.04 |
| C V (%) | 5.01 | 5.13 | 3.34 | 5.25 | 4.89 | 3.31 |
Recovery
The recovery of Human PGF spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.
| Sample Type | Range (%) | Average Recovery (%) |
| Serum (n=5) | 92-105 | 97 |
| EDTA plasma (n=5) | 93-105 | 99 |
| Cell culture media (n=5) | 94-107 | 101 |
Linearity
Samples were spiked with high concentrations of Human PGF 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) | ||
| 1:2 | Range (%) | 94-108 | 98-112 | 92-104 |
| Average (%) | 102 | 105 | 98 | |
| 1:4 | Range (%) | 87-104 | 83-93 | 85-97 |
| Average (%) | 95 | 88 | 90 | |
| 1:8 | Range (%) | 92-106 | 82-94 | 85-97 |
| Average (%) | 97 | 87 | 91 | |
| 1:16 | Range (%) | 91-104 | 84-97 | 85-100 |
| Average (%) | 98 | 91 | 92 |
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 |
- 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.
- Aliquot 100µl of standard solutions into the standard wells.
- Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
- Add 100µl of properly diluted sample (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
- Cover the plate with the sealer provided in the kit and incubate for 90 min at 37°C.
- 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.
- 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.
- Add 100µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37°C.
- Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
- 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.
- Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
- Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.
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