mVEGF QKit SixPak (1 PACK)
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mVEGF QKit SixPak (1 PACK)

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品牌: R&D Systems
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    产品介绍
    产品信息
    产品详情

    Product Summary

    The Quantikine Mouse VEGF Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse VEGF in cell culture supernates, tissue homogenates, mouse serum, and plasma. It contains Sf 21-expressed mouse VEGF and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant mouse VEGF accurately. Results obtained using natural mouse VEGF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse VEGF.

    Recovery

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

    Sample TypeAverage % RecoveryRange %
    Cell Culture Supernates (n=9)10395-114
    EDTA Plasma (n=5)9182-103
    Heparin Plasma (n=4)8987-94
    Serum (n=9)10183-115
    Tissue Homogenates (n=3)9176-107

    Linearity

    To assess the linearity of the assay, samples containing and/or spiked with various concentrations of mouse VEGF in each matrix were diluted with Calibrator Diluent and then assayed.
    Mouse VEGF ELISA Linearity

    Scientific Data

    Mouse VEGF ELISA Standard Curve

    Mouse VEGF Quantikine ELISA Kit Increased production of proinflammatory cytokines and VEGF in the plasma and chondrocytes in high fat diet (HFD)-feeding mice.(a~d) levels of plasma IL-6 (a), TNF-alpha (b), IL-1 beta (c) and VEGF (d) in the C57BL/6 male mice fed with a normal diet (ND) or HFD for 8 weeks. (n = 5, **P < 0.01). (e~h) Relative mRNA expression of IL-6 (e), TNF-alpha (f), IL-1 beta (g) and VEGF (h) in the chondrocytes isolated from the 8-week ND or HFD-feeding mice. (n = 5, **P < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26271607), licensed under a CC-BY license. Not internally tested by R&D Systems.

    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.

    Mouse VEGF Quantikine ELISA Kit Summary

    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), Tissue Homogenates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL)
    Sensitivity
    3 pg/mL
    Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma)
    Specificity
    This kit recognizes both the 164 and 120 amino acid residue forms of mouse VEGF
    Cross-reactivity
    < 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
    Interference
    Interference observed with 1 or more available related molecules.
    背景
    别名
    MVCD1,VAS,vascular endothelial growth factor A,Vascular permeability factor,Vasculotropin,VEGF,VEGFA,VEGF-A,VEGFMGC70609,VPF,VPFvascular endothelial growth factor,Vascular Endothelial Growth Factor
    背景

    Background: VEGF

    Vascular endothelial growth factor (VEGF or VEGF-A), also known as vascular permeability factor (VPF), is a potent mediator of both angiogenesis and vasculogenesis in the fetus and in adults (1-3). It is a member of the PDGF family that is characterized by the presence of eight conserved cysteine residues in a cystine knot structure and the formation of anti-parallel disulfide-linked dimers (4). Alternately spliced isoforms of 120, 164 and 188 amino acids (aa) have been found in mice, while 121, 145, 165, 183, 189, and 206 aa isoforms have been identified in humans (2, 4). In humans, VEGF165 appears to be the most abundant and potent isoform, followed by VEGF121 and VEGF189 (3, 4). The same pattern may exist in mice. Isoforms other than VEGF120 and VEGF121 contain basic heparin-binding regions and are not freely diffusible (4). Mouse VEGF164 shares 97% aa sequence identity with corresponding regions of rat VEGF. It also shares 89% aa sequence identity with human and porcine VEGF, 88% with bovine VEGF, and 90% with feline, equine, and canine VEGF. VEGF is expressed in multiple cells and tissues including skeletal and cardiac muscle (5, 6), hepatocytes (7), osteoblasts (8), neutrophils (9), macrophages (10), keratinocytes (11), brown adipose tissue (12), CD34+ stem cells (13), endothelial cells (14), fibroblasts, and vascular smooth muscle cells (15). VEGF expression is induced by hypoxia and cytokines such as IL-1, IL-6, IL-8, Oncostatin M, and TNF-alpha (3, 4, 9, 16). The isoforms are differentially expressed during development and in the adult (3). 

    VEGF dimers bind to two related receptor tyrosine kinases, VEGF R1 (also called Flt-1) and VEGF R2 (Flk-1/KDR), and induce their homodimerization and autophosphorylation (3, 4, 7, 17, 18). These receptors have seven extracellular immunoglobulin-like domains and an intracellular split tyrosine kinase domain. They are expressed on vascular endothelial cells and a range of non-endothelial cells. Although VEGF affinity is highest for binding to VEGF R1, VEGF R2 appears to be the primary mediator of VEGF angiogenic activity (3, 4). VEGF165 also binds the semaphorin receptor, neuropilin-1, which promotes complex formation with VEGF R2 (19). 
    VEGF is best known for its role in vasculogenesis. During embryogenesis, VEGF regulates the proliferation, migration, and survival of endothelial cells (3, 4), thus regulating blood vessel density and size but playing no role in determining vascular patterns. VEGF promotes bone formation through osteoblast and chondroblast recruitment and is also a monocyte chemoattractant (20-22). In postnatal life, VEGF maintains endothelial cell integrity and is a potent mitogen for micro- and macro-vascular endothelial cells. In adults, VEGF functions mainly in wound healing and the female reproductive cycle (3). In diseased tissues, VEGF promotes vascular permeability. It is thus thought to contribute to tumor metastasis by promoting both extravasation and tumor angiogenesis (23, 24). Various strategies have been employed therapeutically to antagonize VEGF-mediated tumor angiogenesis (25). Circulating VEGF levels correlate with disease activity in autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and systemic lupus erythematosus (26).
    Long Name:
    Vascular Endothelial Growth Factor
    Entrez Gene IDs:
    7422 (Human); 22339 (Mouse); 83785 (Rat); 281572 (Bovine); 403802 (Canine); 493845 (Feline); 30682 (Zebrafish)
    Alternate Names:
    MVCD1; VAS; vascular endothelial growth factor A; Vascular permeability factor; Vasculotropin; VEGF; VEGFA; VEGF-A; VEGFMGC70609; VPF; VPFvascular endothelial growth factor
    制备和贮存
    保存方式

    Preparation and Storage

    Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
    Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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    货号:
    SMMV00
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