Mouse sP-Selectin/CD62P Quantikine ELISA Kit

  (16 citations)     
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Product Details
Assay Procedure
Citations (16)
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  • Assay Type
    Solid Phase Sandwich ELISA
  • Format
    96-well strip plate
  • Assay Length
    4.5 hours
  • Sample Type & Volume Required Per Well
    Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (10 uL)
  • Sensitivity
    0.018 ng/mL
  • Assay Range
    0.3 - 20 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma)
  • Specificity
    Natural and recombinant mouse sP-Selectin
  • Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Product Summary
The Quantikine Mouse soluble P-Selectin (sP-Selectin) Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse sP-Selectin levels in cell culture supernates, serum, and EDTA plasma. It contains NS0-expressed recombinant mouse sP-Selectin and antibodies raised against the recombinant factor. Results obtained for naturally occurring mouse sP-Selectin showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values of natural mouse sP-Selectin.

Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Serum, EDTA Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 32 36 30
Mean 1.27 2.92 12.9 1.27 2.9 12.6
Standard Deviation 0.1 0.23 1.13 0.08 0.24 1.13
CV% 7.9 7.9 8.8 6.3 8.3 9

Recovery

The recovery of mouse sP-Selectin spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 100 89-110
EDTA Plasma (n=6) 94 80-112
Serum (n=10) 98 81-115
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse sP-Selectin in each matrix were diluted with Calibrator Diluent and then assayed.
 P-Selectin/CD62P [HRP]
Product Datasheets

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
  • Entrez Gene IDs:
    6403 (Human); 20344 (Mouse)
  • Alternate Names:
    CD62P antigen; CD62P; FLJ45155; GMP140; GRMP; PADGEM; PADGEMantigen CD62); PSEL; P-Selectin; selectin P (granule membrane protein 140kDa, antigen CD62); SELP
Related Research Areas
Assay Procedure
Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, Control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
  7.   Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
Citations:

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.

16 Citations: Showing 1 - 10
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Species
Sample Type
  1. Sirt3 deficiency does not affect venous thrombosis or NETosis despite mild elevation of intracellular ROS in platelets and neutrophils in mice
    Authors: H Hayashi, D Cherpokova, K Martinod, T Witsch, SL Wong, M Gallant, SM Cifuni, LP Guarente, DD Wagner
    PLoS ONE, 2017;12(12):e0188341.
    Species: Mouse
    Sample Type: Plasma
  2. Molecular magnetic resonance imaging discloses endothelial activation after transient ischaemic attack
    Authors: Aurélien Quenault
    Brain, 2017;140(0):146-157.
    Species: Mouse
    Sample Type: Plasma
  3. Milano summer particulate matter (PM10) triggers lung inflammation and extra pulmonary adverse events in mice.
    Authors: Farina F, Sancini G, Battaglia C, Tinaglia V, Mantecca P, Camatini M, Palestini P
    PLoS ONE, 2013;8(2):e56636.
    Species: Mouse
    Sample Type: Plasma
  4. Early protection from burn-induced acute lung injury by deletion of preprotachykinin-A gene.
    Authors: Sio SW, Moochhala S, Lu J
    Am. J. Respir. Crit. Care Med., 2010;181(1):36-46.
    Species: Mouse
    Sample Type: Tissue Homogenates
  5. Attenuation of transplant arteriosclerosis with clopidogrel is associated with a reduction of infiltrating dendritic cells and macrophages in murine aortic allografts.
    Authors: Abele S, Spriewald BM, Ramsperger-Gleixner M, Wollin M, Hiemann NE, Nieswandt B, Weyand M, Ensminger SM
    Transplantation, 2009;87(2):207-16.
    Species: Mouse
    Sample Type: Serum
  6. The effects of NMDA receptor antagonists over intestinal ischemia/reperfusion injury in rats.
    Authors: Camara-Lemarroy CR, Guzman-de la Garza FJ, Alarcon-Galvan G, Cordero-Perez P, Fernandez-Garza NE
    Eur. J. Pharmacol., 2009;621(1):78-85.
    Species: Rat
    Sample Type: Serum
  7. Biodistribution and safety profile of recombinant adeno-associated virus serotype 6 vectors following intravenous delivery.
    Authors: Stone D, Liu Y, Li ZY, Strauss R, Finn EE, Allen JM, Chamberlain JS, Lieber A
    J. Virol., 2008;82(15):7711-5.
    Species: Mouse
    Sample Type: Serum
  8. High-dose erythropoietin alters platelet reactivity and bleeding time in rodents in contrast to the neuroprotective variant carbamyl-erythropoietin (CEPO).
    Authors: Kirkeby A, Torup L, Bochsen L, Kjalke M, Abel K, Theilgaard-Monch K, Johansson PI, Bjorn SE, Gerwien J, Leist M
    Thromb. Haemost., 2008;99(4):720-8.
    Species: Mouse
    Sample Type: Plasma
  9. Vascular dysfunction in a murine model of severe hemolysis.
    Authors: Frei AC, Guo Y, Jones DW, Pritchard KA, Fagan KA, Hogg N, Wandersee NJ
    Blood, 2008;112(2):398-405.
    Species: Mouse
    Sample Type: Plasma
  10. Reduced plaque formation induced by rosiglitazone in an STZ-diabetes mouse model of atherosclerosis is associated with downregulation of adhesion molecules.
    Authors: Tikellis C, Jandeleit-Dahm KA, Sheehy K, Murphy A, Chin-Dusting J, Kling D, Sebokova E, Cooper ME, Mizrahi J, Woollard KJ
    Atherosclerosis, 2008;199(1):55-64.
    Species: Mouse
    Sample Type: Plasma
  11. Elimination of platelet factor 4 (PF4) from platelets reduces atherosclerosis in C57Bl/6 and apoE-/- mice.
    Authors: Sachais BS, Turrentine T, Dawicki McKenna JM, Rux AH, Rader D, Kowalska MA
    Thromb. Haemost., 2007;98(5):1108-13.
    Species: Mouse
    Sample Type: Platelet-Poor Plasma
  12. Comparison of adenoviruses from species B, C, e, and f after intravenous delivery.
    Authors: Stone D, Liu Y, Li ZY, Tuve S, Strauss R, Lieber A
    Mol. Ther., 2007;15(12):2146-53.
    Species: Mouse
    Sample Type: Serum
  13. Rapid conditional targeted ablation of cells expressing human CD59 in transgenic mice by intermedilysin.
    Authors: Hu W, Ferris SP, Tweten RK, Wu G, Radaeva S, Gao B, Bronson RT, Halperin JA, Qin X
    Nat. Med., 2007;14(1):98-103.
    Species: Mouse
    Sample Type: Plasma
  14. Endogenous hydrogen sulfide regulates leukocyte trafficking in cecal ligation and puncture-induced sepsis.
    Authors: Zhang H, Zhi L, Moochhala SM, Moore PK, Bhatia M
    J. Leukoc. Biol., 2007;82(4):894-905.
    Species: Mouse
    Sample Type: Tissue Homogenates
  15. Cell-based angiopoietin-1 gene therapy for acute lung injury.
    Authors: McCarter SD, Mei SH, Lai PF, Zhang QW, Parker CH, Suen RS, Hood RD, Zhao YD, Deng Y, Han RN, Dumont DJ, Stewart DJ
    Am. J. Respir. Crit. Care Med., 2007;175(10):1014-26.
    Species: Mouse
    Sample Type: Plasma
  16. A fibrinogen deficiency accelerates the initiation of LDL cholesterol-driven atherosclerosis via thrombin generation and platelet activation in genetically predisposed mice.
    Authors: Iwaki T, Sandoval-Cooper MJ, Brechmann M, Ploplis VA, Castellino FJ
    Blood, 2006;107(10):3883-91.
    Species: Mouse
    Sample Type: Plasma
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