Overview

  • Product name

    Mouse PLGF ELISA Kit
    See all PLGF kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 8 4.27%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 6.5%
  • Sample type

    Cell culture supernatant, Serum, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.526 pg/ml
  • Range

    4.69 pg/ml - 300 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 103.8 101.3% - 107.1%
    Cell culture media 98.2 96.2% - 100.6%
    Citrate Plasma 110.7 100.6% - 118.8%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
    Does not react with: Rabbit, Goat, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig
  • Product overview

    PLGF in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PLGF in mouse serum, plasma, and cell culture supernatants.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Sensitivity:
    Samples diluted in Sample Diluent NS – 0.829 pg/mL
    Samples diluted in Sample Diluent 25BS – 0.526 pg/mL
    Samples diluted in Sample Diluent 75BP – 1.559 pg/mL

  • Notes

    Placental growth factor (PLGF) is an angiogenic factor that is variably glycosylated and secreted. The PLGF protein is a 55-60 kDa disulfide linked homodimer and it belongs to the small family of PDGF/VEGF-related angiogenic growth factors, all of which contain 8 conserved cysteines. Due to alternative splicing, there are three human PLGF mRNAs encoding human PLGF-1, -2, and -3 isoform precursors. There is a unique 21 aa insert at the C-terminus of the PLGF-2 isoform and this isoform is the heparin binding form of PLGF. In mouse tissues there is currently only one identified PLGF mRNA form encoding the equivalent of human PLGF-2. Mouse PLGF is a 158 aa polypeptide that contains an 18 aa signal sequence and a 140 aa mature segment. Human and rat PLGF share 66% and 91% aa sequence identity, respectively, to mouse PLGF.

    Homodimeric mouse PLGF binds the tyrosine kinase receptor VEGF R1/Flt-1. This receptor is a 180 kDa type I transmembrane glycoprotein with high affinity for PLGF. In addition to forming homodimers, PLGF also forms heterodimers with VEGF-A isoforms, forming bioactive molecules. When heterodimerized with VEGF, PLGF will activate VEGF R2.

    A limited number of functions have been directly attributed to PLGF. It is known to induce monocyte activation and to provide a stimulus for monocyte migration. PLGF also upregulates VEGF production in PBMCs and induces tissue factor secretion in both monocytes and HUVECs. Furthermore, PLGF is also reported to serve as a growth and chemotactic factor for endothelial cells. Finally, PLGF is known to be expressed by trophoblasts and may protect against growth factor withdrawal-induced trophoblast apoptosis.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse PLGF Capture Antibody 1 x 600µl
    10X Mouse PLGF Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Mouse PLGF Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent 25BS 1 x 20ml
    Sample Diluent 75BP 1 x 20ml
    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

  • Function

    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.
  • Tissue specificity

    While the three isoforms are present in most placental tissues, PlGF-2 is specific to early (8 week) placenta and only PlGF-1 is found in the colon and mammary carcinomas.
  • Sequence similarities

    Belongs to the PDGF/VEGF growth factor family.
  • Domain

    Isoform PlGF-2 contains a basic insert which acts as a cell retention signal.
  • Post-translational
    modifications

    N-glycosylated.
  • Cellular localization

    Secreted. The three isoforms are secreted but PlGF-2 appears to remain cell attached unless released by heparin.
  • Information by UniProt
  • Alternative names

    • AI854365
    • D12S1900
    • MGC93298
    • Pgf
    • PGFL
    • PIGF
    • Placenta growth factor
    • Placental growth factor
    • Placental growth factor vascular endothelial growth factor related protein
    • PlGF
    • PlGF 2
    • PLGF_HUMAN
    • PlGF2
    • SHGC 10760
    • Vascular endothelial growth factor related protein
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab197748 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • 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.

     

  • Standard curve comparison between mouse PLGF SimpleStep ELISA® kit and traditional ELISA kit from leading competitor. SimpleStep ELISA kit shows a 2-fold sensitivity.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD) are graphed.

  • L929, J774A.1 and RAW264.7 cells were cultured in HGDMEM with 10% fetal calf serum or Horse serum, 2mM L-glutamine and 100 µg/mL Kanamycin. L929 and J774A.1 cells were treated for 72 hours in the presence and absence of 1.5% PHA and 10ng/mL of PMA. RAW264.7 cells were starved for 24 hours and treated in the presence and absence of 5 µg/mL of LPS. Samples were loaded on the plate undiluted and the concentrations of PLGF were interpolated from a calibration curve diluted in NS. The mean PLGF concentration was determined to be 178.9 pg/mL in stimulated L929 cells, 15.8 pg/mL in unstimulated L929 cells, 32.8 pg/mL in stimulated J774A.1 cells, 0.4 pg/mL in unstimulated J774A.1 cells, 4.4 pg/mL in stimulated RAW264.7 cells, and 2.3 pg/mL in unstimulated RAW264.7 cells.

Protocols

References

ab197748 has not yet been referenced specifically in any publications.

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