Human PlGF DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human PlGF. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.
The components listed above may be purchased separately:
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4
Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: R&D Systems (Catalog # DY990)
Plate Sealers: ELISA Plate Sealers (Catalog # DY992)
Scientific Data
Product Datasheets
Preparation and Storage
Background: PlGF
PlGF (placenta growth factor) is a homodimeric growth factor that competes with VEGF for binding to VEGF R1/Flt-1. It therefore increases the availability of VEGF to bind to VEGF R2/KDR/Flk-1 and trigger angiogenesis. It can form heterodimers with some forms of VEGF and decrease the angiogenic effect of VEGF on VEGF R2. Circulating PlGF levels increase during pregnancy, reaching a peak in mid-gestation; this increase is attenuated in preeclampsia. PlGF induces monocyte activation and migration as well as production of inflammatory cytokines and VEGF. These activities facilitate wound, bone fracture, and cardiac repair, but also contribute to inflammation in active sickle cell disease and atherosclerosis. PlGF can also inhibit TIMP3 expression in the spleen, leading to immune triggering of hypertension.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human PlGF DuoSet ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
17
Citations: Showing 1 - 10
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Platelet and mitochondrial RNA is decreased in plasma-derived extracellular vesicles in women with preeclampsia-an exploratory study
Authors: Lekva, T;Sundaram, AYF;Roland, MCP;Åsheim, J;Michelsen, AE;Norwitz, ER;Aukrust, P;Gilfillan, GD;Ueland, T;
BMC medicine
Species: Human
Sample Types: Plasma
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Combination of Maternal Serum ESM-1 and PLGF with Uterine Artery Doppler PI for Predicting Preeclampsia
Authors: X Xie, D Chen, X Yang, Y Cao, Y Guo, W Cheng
Journal of Clinical Medicine, 2023-01-06;12(2):.
Species: Human
Sample Types: Serum
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Hydroxychloroquine reduces soluble Flt-1 secretion from human cytotrophoblast, but does not mitigate markers of endothelial dysfunction in vitro
Authors: E Kadife, N Hannan, A Harper, N Binder, S Beard, FC Brownfoot
PLoS ONE, 2022-11-23;17(11):e0271560.
Species: Human
Sample Types: Cell Culture Supernates
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Reduced Interleukin-17-Expressing Cells in Cutaneous Melanoma
Authors: A Tosi, L Nardinocch, ML Carbone, L Capriotti, E Pagani, S Mastroeni, C Fortes, F Scopelliti, C Cattani, F Passarelli, A Rosato, S D'Atri, CM Failla, A Cavani
Biomedicines, 2021-12-16;9(12):.
Species: Human
Sample Types: Cell Culture Supernates
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A prospective cohort study providing insights for markers of adverse pregnancy outcome in older mothers
Authors: SC Lean, RL Jones, SA Roberts, AEP Heazell
BMC pregnancy and childbirth, 2021-10-20;21(1):706.
Species: Human
Sample Types: Serum
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Hyperglycaemia up-regulates placental growth factor (PlGF) expression and secretion in endothelial cells via suppression of PI3 kinase-Akt signalling and activation of FOXO1
Authors: S Sissaoui, S Egginton, L Ting, A Ahmed, PW Hewett
Scientific Reports, 2021-08-11;11(1):16344.
Species: Human
Sample Types: Cell Culture Supernates
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Fluid shear stress regulates placental growth factor expression via heme oxygenase 1 and iron
Authors: NA Rashdan, B Zhai, PC Lovern
Scientific Reports, 2021-07-21;11(1):14912.
Species: Human
Sample Types: Cell Culture Supernates
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No association between early antiretroviral therapy during pregnancy and plasma levels of angiogenic factors: a cohort study
Authors: A Djeha, S Girard, H Trottier, F Kakkar, H Soudeyns, M Boucher, N Lapointe, I Boucoiran
BMC Pregnancy Childbirth, 2019-12-09;19(1):482.
Species: Human
Sample Types: Serum
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Sulfasalazine reduces placental secretion of antiangiogenic factors, up-regulates the secretion of placental growth factor and rescues endothelial dysfunction
Authors: FC Brownfoot, NJ Hannan, P Cannon, V Nguyen, R Hastie, LJ Parry, S Senadheera, L Tuohey, S Tong, TJ Kaitu'u-Li
EBioMedicine, 2019-02-26;0(0):.
Species: Human
Sample Types: Tissue Culture Supernates
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Placental Dysfunction Underlies Increased Risk of Fetal Growth Restriction and Stillbirth in Advanced Maternal Age Women
Authors: SC Lean, AEP Heazell, MR Dilworth, TA Mills, RL Jones
Sci Rep, 2017-08-29;7(1):9677.
Species: Human
Sample Types: Serum
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Angiogenic proteins, placental weight and perinatal outcomes among pregnant women in Tanzania
PLoS ONE, 2016-12-09;11(12):e0167716.
Species: Human
Sample Types: Plasma
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Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
PLoS ONE, 2015-08-06;10(8):e0134619.
Species: Human
Sample Types: Plasma
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Serum concentrations of soluble Flt-1 are decreased among women with a viable fetus and no symptoms of miscarriage destined for pregnancy loss.
Authors: Kaitu'u-Lino TJ, Whitehead CL, Ngian GL, Permezel M, Tong S
PLoS ONE, 2012-02-28;7(2):e32509.
Species: Human
Sample Types: Serum
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Periodic assessment of plasma sFlt-1 and PlGF concentrations and its association with placental morphometry in gestational hypertension (GH) - a prospective follow-up study.
Authors: Jeevaratnam K, Nadarajah VD, Judson JP
BMC Pregnancy Childbirth, 2010-09-28;10(0):58.
Species: Human
Sample Types: Plasma
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Secretion of angiogenic growth factors by villous cytotrophoblast and extravillous trophoblast in early human pregnancy.
Authors: Lash GE, Naruse K, Innes BA, Robson SC, Searle RF, Bulmer JN
Placenta, 2010-03-24;31(6):545-8.
Species: Human
Sample Types: Cell Culture Supernates
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Impact of heterogeneity of human peripheral blood monocyte subsets on myocardial salvage in patients with primary acute myocardial infarction.
Authors: Tsujioka H, Imanishi T, Ikejima H, Kuroi A, Takarada S, Tanimoto T, Kitabata H, Okochi K, Arita Y, Ishibashi K, Komukai K, Kataiwa H, Nakamura N, Hirata K, Tanaka A, Akasaka T
J. Am. Coll. Cardiol., 2009-07-07;54(2):130-8.
Species: Human
Sample Types: Plasma
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Hypoxia-induced increase in soluble Flt-1 production correlates with enhanced oxidative stress in trophoblast cells from the human placenta.
Authors: Li H, Gu B, Zhang Y, Lewis DF, Wang Y
Placenta, 2005-02-01;26(2):210-7.
Species: Human
Sample Types: Cell Culture Supernates
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We assayed the supernatant from cultured human thyroid explants
