Human P-Selectin/CD62P 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 P-Selectin/CD62P. 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: P-Selectin/CD62P
The Selectin family is comprised of three members, E-Selectin, L-Selectin, and P-Selectin. E-Selectin [endothelial leukocyte adhesion molecule-1 (ELAM-1), CD62E] is transiently expressed on vascular endothelial cells in response to IL-1 beta and TNF-alpha. The human and rat proteins share approximately 67% amino acid sequence identity. The mouse and rat proteins share approximately 78% amino acid sequence identity. L-Selectin (Leukocyte Selectin, LAM-1, LECAM-1, LECCAM-1, TQ1, Leu-8, MEL-14 antigen, DREG, lymph node homing receptor, CD62L) is expressed constitutively on a wide variety of leukocytes. Two forms of L-Selectin have been reported, apparently arising as a result of post-translational modifications. Human and mouse L-Selectin share 76% amino acid sequence identity. Human P-Selectin (GMP-140, LECAM-3, PADGEM, CD62P) is expressed by activated platelets and endothelial cells. The extracellular domains of human and mouse P-Selectin share approximately 73% amino acid sequence identity.
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 P-Selectin/CD62P 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.
20
Citations: Showing 1 - 10
Filter your results:
Filter by:
-
Baseline circulating biomarkers, their changes, and subsequent suicidal ideation and depression severity at 6 months: A prospective analysis in patients with mood disorders
Authors: Lengvenyte, A;Cognasse, F;Hamzeh-Cognasse, H;Sénèque, M;Strumila, R;Olié, E;Courtet, P;
Psychoneuroendocrinology
Species: Human
Sample Types: Plasma
-
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
-
CD36 mediates SARS-CoV-2-envelope-protein-induced platelet activation and thrombosis
Authors: Tang, Z;Xu, Y;Tan, Y;Shi, H;Jin, P;Li, Y;Teng, J;Liu, H;Pan, H;Hu, Q;Cheng, X;Ye, J;Su, Y;Sun, Y;Meng, J;Zhou, Z;Chi, H;Wang, X;Liu, J;Lu, Y;Liu, F;Dai, J;Yang, C;Chen, S;Liu, T;
Nature communications
Species: Human
Sample Types: Plasma
-
Exploring dysregulated immune response genes and endothelial dysfunction biomarkers as predictors of severe COVID-19
Authors: Reis-Goes, FS;Silva, NN;Gondim, TM;Figueiredo, RG;Evangelista, GAO;Marchioro, SB;Costa, RS;Torres, AJL;Meyer, RJ;Trindade, SC;Fortuna, V;
International immunopharmacology
Species: Human
Sample Types: Plasma
-
Association of PCSK9 with inflammation and platelet activation markers and recurrent cardiovascular risks in STEMI patients undergoing primary PCI with or without diabetes
Authors: L Song, X Zhao, R Chen, J Li, J Zhou, C Liu, P Zhou, Y Wang, Y Chen, H Zhao, H Yan
Cardiovascular Diabetology, 2022-05-20;21(1):80.
Species: Human
Sample Types: Plasma
-
Prediction of emergency cerclage outcomes in women with cervical insufficiency: The role of inflammatory, angiogenic, and extracellular matrix-related proteins in amniotic fluid
Authors: KN Lee, KH Park, YM Kim, I Cho, TE Kim
PLoS ONE, 2022-05-10;17(5):e0268291.
Species: Human
Sample Types: Amniotic Fluid
-
The effect of transdermal gender-affirming hormone therapy on markers of inflammation and hemostasis
Authors: MH Schutte, R Kleemann, NM Nota, CM Wiepjes, JM Snabel, G T'Sjoen, A Thijs, M den Heijer
PLoS ONE, 2022-03-15;17(3):e0261312.
Species: Human
Sample Types: Plasma
-
The effect of moderate wine consumption on cytokine secretion by peripheral blood mononuclear cells: A randomized clinical study in coronary heart disease patients
Authors: E Fragopoulo, C Argyrou, M Detopoulou, S Tsitsou, S Seremeti, M Yannakouli, S Antonopoul, G Kolovou, P Kalogeropo
Cytokine, 2021-07-08;146(0):155629.
Species: Human
Sample Types: Plasma
-
Elevation of C-reactive protein, P-selectin and Resistin as potential inflammatory biomarkers of urogenital Schistosomiasis exposure in preschool children
Authors: TN Chimponda, C Mushayi, DNM Osakunor, A Vengesai, E Enwono, S Amanfo, J Murray, C Tshuma, F Mutapi, T Mduluza
BMC Infect. Dis., 2019-12-19;19(1):1071.
Species: Human
Sample Types: Serum
-
Blood inflammatory and endothelial markers in women with von Willebrand disease
Authors: I Govorov, K Bremme, A Larsson, M Holmström, E Komlichenk, R Chaireti, M Mints
PLoS ONE, 2019-01-10;14(1):e0210544.
Species: Human
Sample Types: Plasma
-
Preemptively and non-preemptively transplanted patients show a comparable hypercoagulable state prior to kidney transplantation compared to living kidney donors
Authors: GJ Nieuwenhui, TAJ van den Be, SJL Bakker, MC van den He, MMRF Struys, T Lisman, RA Pol
PLoS ONE, 2018-07-16;13(7):e0200537.
Species: Human
Sample Types: Plasma
-
Biomarkers of Host Response Predict Primary End-Point Radiological Pneumonia in Tanzanian Children with Clinical Pneumonia: A Prospective Cohort Study.
Authors: Erdman L, D'Acremont V, Hayford K, Rajwans N, Kilowoko M, Kyungu E, Hongoa P, Alamo L, Streiner D, Genton B, Kain K
PLoS ONE, 2015-09-14;10(9):e0137592.
Species: Human
Sample Types: Plasma
-
Highly invasive melanoma cells activate the vascular endothelium via an MMP-2/integrin alphavbeta5-induced secretion of VEGF-A.
Am. J. Pathol., 2012-05-31;181(2):693-705.
Species: Human
Sample Types: Cell Culture Supernates
-
Up-regulation of platelet activation in hemophilia A.
Authors: van Bladel ER, Roest M
Haematologica, 2011-03-21;0(0):.
Species: Human
Sample Types: Plasma
-
Combinations of Host Biomarkers Predict Mortality among Ugandan Children with Severe Malaria: A Retrospective Case-Control Study.
Authors: Erdman LK, Dhabangi A, Musoke C, Conroy AL, Hawkes M, Higgins S, Rajwans N, Wolofsky KT, Streiner DL, Liles WC, Cserti-Gazdewich CM, Kain KC
PLoS ONE, 2011-02-25;6(2):e17440.
Species: Human
Sample Types: Plasma
-
Thrombocytopenia in early malaria is associated with GP1b shedding in absence of systemic platelet activation and consumptive coagulopathy.
Authors: de Mast Q, de Groot PG, van Heerde WL, Roestenberg M, van Velzen JF, Verbruggen B, Roest M, McCall M, Nieman AE, Westendorp J, Syafruddin D, Fijnheer R, van Dongen-Lases EC, Sauerwein RW, van der Ven AJ
Br. J. Haematol., 2010-10-19;151(5):495-503.
Species: Human
Sample Types: Plasma
-
High platelet reactivity is associated with myocardial infarction in premenopausal women: a population-based case-control study.
Authors: Snoep JD, Roest M, Barendrecht AD
J. Thromb. Haemost., 2010-01-30;8(5):906-13.
Species: Human
Sample Types: Plasma
-
Delay of acute intracellular pH recovery after acidosis decreases endothelial cell activation.
Authors: Huck V, Niemeyer A, Goerge T, Schnaeker EM, Ossig R, Rogge P, Schneider MF, Oberleithner H, Schneider SW
J. Cell. Physiol., 2007-05-01;211(2):399-409.
Species: Human
Sample Types: Cell Culture Supernates
-
Cardiovascular risk factors are reduced with a low dose of acarbose in obese patients with polycystic ovary syndrome.
Authors: Araujo Penna I, Canella PR, Vieira CS, Silva de Sa MF, dos Reis RM, Ferriani RA
Fertil. Steril., 2007-04-06;88(2):519-22.
Species: Human
Sample Types: Serum
-
Plasma cytokine and P-selectin levels in advanced malignancy: prognostic value and impact of low-molecular weight heparin administration.
Authors: Di Nisio M, Niers TM, Reitsma PH, Buller HR
Cancer, 2005-11-15;104(10):2275-81.
Species: Human
Sample Types: Plasma
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human P-Selectin/CD62P DuoSet ELISA
Average Rating: 5 (Based on 1 Review)
Have you used Human P-Selectin/CD62P DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by:
