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    products/elisa/human-cd31-elisa-kitpecam-ab190814.pdf

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RecombinantSimpleStep

Human CD31 ELISA Kit (PECAM) (ab190814)

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  • SDS
  • Protocol Booklet
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Other - Human CD31 ELISA Kit (PECAM) (ab190814)
  • Human CD31 Standard Curve Comparison
  • Human CD31 Standard Curve Comparison
  • Example of human CD31 standard curve in Sample Diluent NS + Enhancer.
  • Example of human CD31 standard curve in 1X Cell Extraction Buffer PTR + Enhancer.
  • Interpolated concentrations of native CD31 in human serum and plasma samples.
  • . Interpolated concentrations of native CD31 in human saliva, urine, and THP-1 cell culture supernatant samples.
  • Interpolated concentrations of native CD31 in THP-1 cell extract samples based on a 125 µg/mL extract load.
  • Serum from ten individual healthy human female donors was measured in duplicate.
  • Sandwich ELISA - Human CD31 ELISA Kit (PECAM) (ab190814)

Key features and details

  • One-wash 90 minute protocol
  • Sensitivity: 44 pg/ml
  • Range: 226.6 pg/ml - 14500 pg/ml
  • Sample type: Cell culture extracts, Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Saliva, Serum, Tissue Extracts, Urine
  • Detection method: Colorimetric
  • Assay type: Sandwich (quantitative)
  • Reacts with: Human, Rhesus monkey

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Overview

  • Product name

    Human CD31 ELISA Kit (PECAM)
    See all CD31 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    cell extract 8 3.4%
    Inter-assay
    Sample n Mean SD CV%
    cell extract 3 3.3%
  • Sample type

    Cell culture supernatant, Saliva, Urine, Serum, Cell culture extracts, Tissue Extracts, Hep Plasma, EDTA Plasma, Cit plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    44 pg/ml
  • Range

    226.6 pg/ml - 14500 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Urine 80 76% - 84%
    Serum 113 109% - 119%
    Hep Plasma 74 71% - 80%
    EDTA Plasma 103 102% - 106%
    Cit plasma 91 88% - 95%
  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human, Rhesus monkey
    Does not react with: Cow
  • Product overview

    Human CD31 ELISA Kit (PECAM) (ab190814) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of CD31 (PECAM) protein in cell culture extracts, cell culture supernatant, cit plasma, edta plasma, hep plasma, saliva, serum, tissue extracts, and urine. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human CD31 (PECAM) with 44 pg/ml sensitivity.


    SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:


            - Single-wash protocol reduces assay time to 90 minutes or less
            - High sensitivity, specificity and reproducibility from superior antibodies
            - Fully validated in biological samples
            - 96-wells plate breakable into 12 x 8 wells strips


    A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.

  • Notes

    CD31, also known as PECAM-1, is a cell adhesion molecule which is required for leukocyte trans-endothelial migration (TEM) under most inflammatory conditions. CD31 is a 130-kDa glycoprotein constitutively expressed on endothelial cells, platelets, neutrophils, monocytes, lymphocytes and basophils. CD31 molecules is expressed in large amounts in the intracellular junctions of endothelial cells. Experiments using antibody results in blocking CD31 homophilic interaction and inhibition of leukocytes transmigration. Expression of CD31 on the cell surface is regulated by receptor shedding resulting in generation of a soluble 100-kDa extracellular domain form and a truncated 28-kDa fragment containing the transmembrane and cytoplasmic domain of CD31.

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Human CD31 Capture Antibody 1 x 600µl
    10X Human CD31 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 2 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent 4BI 1 x 6ml
    Denaturant 1 x 500µl
    Human CD31 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS (ab193972) 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

    • Cardiovascular
    • Blood
    • Platelets
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Cell Adhesion Molecules
    • Endothelial
    • Cardiovascular
    • Angiogenesis
    • Angiogenic Factors
    • Immunology
    • Cell Type Markers
    • CD
    • Adhesion
    • Stem Cells
    • Mesenchymal Stem Cells
    • Surface Molecules
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Other
    • Cancer
    • Tumor immunology
    • CD markers
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Cell adhesion molecules
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Neutrophil Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Thrombocytic Lineage
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    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers
  • Function

    Induces susceptibility to atherosclerosis (By similarity). Cell adhesion molecule which is required for leukocyte transendothelial migration (TEM) under most inflammatory conditions. Tyr-690 plays a critical role in TEM and is required for efficient trafficking of PECAM1 to and from the lateral border recycling compartment (LBRC) and is also essential for the LBRC membrane to be targeted around migrating leukocytes. Prevents phagocyte ingestion of closely apposed viable cells by transmitting 'detachment' signals, and changes function on apoptosis, promoting tethering of dying cells to phagocytes (the encounter of a viable cell with a phagocyte via the homophilic interaction of PECAM1 on both cell surfaces leads to the viable cell's active repulsion from the phagocyte. During apoptosis, the inside-out signaling of PECAM1 is somehow disabled so that the apoptotic cell does not actively reject the phagocyte anymore. The lack of this repulsion signal together with the interaction of the eat-me signals and their respective receptors causes the attachment of the apoptotic cell to the phagocyte, thus triggering the process of engulfment). Isoform Delta15 is unable to protect against apoptosis. Modulates BDKRB2 activation. Regulates bradykinin- and hyperosmotic shock-induced ERK1/2 activation in human umbilical cord vein cells (HUVEC).
  • Tissue specificity

    Expressed on platelets and leukocytes and is primarily concentrated at the borders between endothelial cells. Isoform Long predominates in all tissues examined. Isoform Delta12 is detected only in trachea. Isoform Delta14-15 is only detected in lung. Isoform Delta14 is detected in all tissues examined with the strongest expression in heart. Isoform Delta15 is expressed in brain, testis, ovary, cell surface of platelets, human umbilical vein endothelial cells (HUVECs), Jurkat T-cell leukemia, human erythroleukemia (HEL) and U937 histiocytic lymphoma cell lines (at protein level).
  • Sequence similarities

    Contains 6 Ig-like C2-type (immunoglobulin-like) domains.
  • Domain

    The Ig-like C2-type domains 2 and 3 contribute to formation of the complex with BDKRB2 and in regulation of its activity.
  • Post-translational
    modifications

    Phosphorylated on Ser and Tyr residues after cellular activation. In endothelial cells Fyn mediates mechanical-force (stretch or pull) induced tyrosine phosphorylation.
  • Cellular localization

    Membrane. Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells and Cell junction. Localizes to the lateral border recycling compartment (LBRC) and recycles from the LBRC to the junction in resting endothelial cells.
  • Target information above from: UniProt accession P16284 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Alternative names

    • Adhesion molecule
    • CD31
    • CD31 antigen
    • CD31 EndoCAM
    • EndoCAM
    • FLJ34100
    • FLJ58394
    • GPIIA
    • GPIIA'
    • PECA1
    • PECA1_HUMAN
    • PECAM
    • Pecam 1
    • PECAM 1 CD31 EndoCAM
    • PECAM-1
    • Pecam1
    • Platelet and endothelial cell adhesion molecule 1
    • Platelet endothelial cell adhesion molecule
    • Platelet/endothelial cell adhesion molecule 1
    see all
  • Database links

    • Entrez Gene: 5175 Human
    • Omim: 173445 Human
    • SwissProt: P16284 Human
    • Unigene: 376675 Human
    • Unigene: 514412 Human

    Associated products

    • Alternative Versions

      • Human CD31 Antibody Pair - BSA and Azide free (PECAM) (ab256599)
    • ELISA kits

      • Human CD31 ELISA Kit (PECAM1) (ab45910)

    Images

    • Other - Human CD31 ELISA Kit (PECAM) (ab190814)
      Other - Human CD31 ELISA Kit (PECAM) (ab190814)

      SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

       

    • Human CD31 Standard Curve Comparison
      Human CD31 Standard Curve Comparison

      Standard Curve comparison between human CD31 SimpleStep ELISA kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows increased sensitivity.

    • Human CD31 Standard Curve Comparison
      Human CD31 Standard Curve Comparison

      Standard Curve comparison between human CD31 new SimpleStep ELISA kit and previous ELISA kit. The new SimpleStep ELISA kit shows increased sensitivity.

    • Example of human CD31 standard curve in Sample Diluent NS + Enhancer.
      Example of human CD31 standard curve in Sample Diluent NS + Enhancer.

      The CD31 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

    • Example of human CD31 standard curve in 1X Cell Extraction Buffer PTR + Enhancer.
      Example of human CD31 standard curve in 1X Cell Extraction Buffer PTR + Enhancer.

      The CD31 standard curve was prepared as described in Section 10. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.

       

    • Interpolated concentrations of native CD31 in human serum and plasma samples.
      Interpolated concentrations of native CD31 in human serum and plasma samples.

      The concentrations of CD31 were measured in duplicates, interpolated from the CD31 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 36%, plasma (citrate) 9%, plasma (heparin) 9% and plasma (EDTA) 9%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD31 concentration was determined to be 6.60 ng/mL in neat serum, 5.04 ng/mL in neat plasma (citrate), 5.67 ng/mL in neat plasma (heparin), and 6.44 ng/mL in neat plasma (EDTA).

    • . Interpolated concentrations of native CD31 in human saliva, urine, and THP-1 cell culture supernatant samples.
      . Interpolated concentrations of native CD31 in human saliva, urine, and THP-1 cell culture supernatant samples.

      THP-1 cells were cultured in the presence (stimulated) or absence (mock) of 5 µg/mL LPS and 50nM PMA for 24 hours. The concentrations of CD31 were measured in duplicates, interpolated from the CD31 standard curves and corrected for sample dilution. Undiluted samples are as follows: saliva 36%, urine 36%, THP-1 stimulated supernatant 18%, and THP-1 mock supernatant 18%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD31 concentration was determined to be 6.85 ng/mL in neat saliva, 0.81 ng/mL in neat urine, 2.27 ng/mL in neat stimulated THP-1 supernatant, and 0.64 ng/mL in neat mock THP-1 supernatant.

    • Interpolated concentrations of native CD31 in THP-1 cell extract samples based on a 125 µg/mL extract load.
      Interpolated concentrations of native CD31 in THP-1 cell extract samples based on a 125 µg/mL extract load.

      THP-1 cells were cultured in the presence (stimulated) or absence (unstimulated) of 5 µg/mL LPS and 50nM PMA for 24 hours. The concentrations of CD31 were measured in duplicate and interpolated from the CD31 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD31 concentration was determined to be 16.0 ng/mL in stimulated THP-1 cell extract and 12.5 ng/mL in unstimulated THP-1 cell extract.

       

    • Serum from ten individual healthy human female donors was measured in duplicate.
      Serum from ten individual healthy human female donors was measured in duplicate.

      Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean CD31 concentration was determined to be 6.34 ng/mL with a range of 4.39 – 11.1 ng/mL

    • Sandwich ELISA - Human CD31 ELISA Kit (PECAM) (ab190814)
      Sandwich ELISA - Human CD31 ELISA Kit (PECAM) (ab190814)
      To learn more about the advantages of recombinant antibodies see here.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab190814? Please let us know so that we can cite the reference in this datasheet.

    ab190814 has been referenced in 1 publication.

    • Meng D  et al. Effects of VEGFR1+ hematopoietic progenitor cells on pre-metastatic niche formation and in vivo metastasis of breast cancer cells. J Cancer Res Clin Oncol 145:411-427 (2019). PubMed: 30483898

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