Culture supernatant was collected from each growth condition, and keratinocyte growth factor (KGF-7) concentration was determined using the Quantikine human KGF/FGF-7 Immunoassay (R&D Systems Inc., Minneapolis, MN, USA). An ExoQuick-TC® ULTRA EV Kit and ExoELISA-ULTRA CD63 Kit (System Biosciences, Palo Alto, CA, USA) were used for extracellular vesicle isolation and quantification, respectively. For the semiquantitative detection of 23 human proteins in cell culture media, the human cytokine antibody array C1 (AAH-CYT-1; RayBiotech, Norcross, GA, USA), was used (Supplementary Figure S1 ). All determinations were performed according to the manufacturer’s instructions.
Quantikine human kgf fgf 7 immunoassay
Manufactured by R&D Systems
Sourced in United States
The Quantikine human KGF/FGF-7 Immunoassay is a quantitative sandwich enzyme immunoassay designed to measure human keratinocyte growth factor (KGF/FGF-7) levels in cell culture supernates, serum, and plasma. The assay employs an antibody specific for human KGF/FGF-7 coated on a microplate. Standards and samples are pipetted into the wells, and any KGF/FGF-7 present is bound by the immobilized antibody.
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2 protocols using quantikine human kgf fgf 7 immunoassay
1
Extracellular Vesicle Isolation and Quantification
2
Protein Expression and Cytokine Analysis
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Whole cell lysates were prepared using MCL1 lysis buffer in the presence of protease and phosphatase cocktail inhibitor (Sigma-Aldrich, St Louis, MO) following the manufacter's protocol. Protein lysates were subjected to SDS-PAGE on 10% polyacrylamide gel and transferred onto PVDF membranes. Membranes were probed for the protein levels of E-cadherin, and α-tubulin as loading control, using speci c primary antibodies (#4065, Cell Signaling Tech. Inc., Danvers, MA, and #sc-5286, Santa Cruz Biotechnology, Inc., Dallas, TX, respectively). Speci c bands for each protein were visualized by WesternBright™ kit (Advansta, San Jose, CA).
Enzyme-linked Immunosorbent Assay (ELISA) and human cytokine array.
Culture supernatant was collected from each growth condition, and keratinocyte growth factor (KGF-7) concentration was determined using Quantikine human KGF/FGF-7 Immunoassay (R&D Systems Inc., Minneapolis, MN). ExoQuick-TC® ULTRA EV Kit and ExoELISA-ULTRA CD63 Kit (System Biosciences, Palo Alto, CA) were used for extracellular vesicles isolation and quanti cation, respectively. For the semiquantitative detection of 23 human protein in cell culture media, the human cytokine antibody array C1 (AAH-CYT-1; RayBiotech, Norcross, GA), was used (Supplementary Figure 2). All determinations were performed according to the manufacturer's instructions.
Enzyme-linked Immunosorbent Assay (ELISA) and human cytokine array.
Culture supernatant was collected from each growth condition, and keratinocyte growth factor (KGF-7) concentration was determined using Quantikine human KGF/FGF-7 Immunoassay (R&D Systems Inc., Minneapolis, MN). ExoQuick-TC® ULTRA EV Kit and ExoELISA-ULTRA CD63 Kit (System Biosciences, Palo Alto, CA) were used for extracellular vesicles isolation and quanti cation, respectively. For the semiquantitative detection of 23 human protein in cell culture media, the human cytokine antibody array C1 (AAH-CYT-1; RayBiotech, Norcross, GA), was used (Supplementary Figure 2). All determinations were performed according to the manufacturer's instructions.
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