ELISAs were performed following manufacturers’ instructions using the Human/Mouse/Rat Activin-A Quantikine ELISA Kit (#DAC00B, R&D Systems), the Human IL-6 Quantikine ELISA Kit (#D6050, R&D Systems) and the Human G-CSF Quantikine ELISA Kit (#DCS50, R&D Systems).
Human g csf quantikine elisa kit
Manufactured by R&D Systems
Sourced in United States
The Human G-CSF Quantikine ELISA Kit is a quantitative sandwich enzyme immunoassay designed to measure human Granulocyte Colony-Stimulating Factor (G-CSF) levels in cell culture supernates, serum, and plasma.
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Lab products found in correlation
Human il 6 quantikine elisa kit, by R&D Systems (1 mentions)
Human mouse rat activin a quantikine elisa kit, by R&D Systems (1 mentions)
Imark microplate reader, by Bio-Rad (1 mentions)
Dnase 1, by Roche (1 mentions)
Human mpo elisa kit, by Hycult Biotech (1 mentions)
Human cell death detection elisaplus, by Roche (1 mentions)
Human gm csf quantikine elisa kit, by R&D Systems (1 mentions)
Gentlemacs dissociator, by Miltenyi Biotec (1 mentions)
Mouse quantikine elisa kit, by R&D Systems (1 mentions)
Endothelial cell growth medium 2, by PromoCell (1 mentions)
6 protocols using human g csf quantikine elisa kit
1
Quantitative ELISA Analyses of Activin-A, IL-6, and G-CSF
2
Quantifying Human G-CSF Levels
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The concentrations of human G-CSF were measured using a Human G-CSF Quantikine ELISA Kit (R&D Systems; #DCS50). Absorbance values were measured using a microplate reader (iMark Microplate Reader; Bio-Rad Laboratories, Inc., Hercules, CA).
3
Quantification of NET-associated Biomarkers
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The concentrations of NE and MPO were measured in sera and plasma by sandwich ELISA, utilizing, respectively, the Elastase/a1-PI Complex ELISA Kit (Calbiochem) and the human MPO ELISA Kit. Histone/DNA complexes in sera and plasma were measured using the Human Cell Death Detection ELISAPLUS (Roche Diagnostics); nucleosomes in cell culture supernatants were detected similarly after incubation with DNase I (10 U for 5 min) (Roche Diagnostics). To identify NET-associated MPO/DNA complexes, a modified capture ELISA was utilized (27 (link)). NET-associated MPO in culture supernatant was captured using the coated 96-well plate of the human MPO ELISA Kit (Hycult Biotech), and the NET-associated DNA backbone was detected using the anti-DNA-POD antibody of the Human Cell Death Detection ELISAPLUS (Roche Diagnostics). G-CSF serum and plasma protein concentrations were assessed with the Human G-CSF Quantikine ELISA Kit (R&D Systems).
4
Plasma G-CSF Quantification by ELISA
5
Quantification of G-CSF and GM-CSF
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The concentration of G-CSF or GM-CSF in all samples was evaluated using an ELISA kit (R&D Systems, USA, Human G-CSF Quantikine ELISA Kit and Human GM-CSF Quantikine ELISA Kit), following the manufacturer’s instructions. To ensure measurements within the detection range of the ELISA kits, GM-CSF samples were diluted at a 1:5 ratio using the dilution buffer provided with the ELISA kit. Each sample was analyzed in duplicate during the ELISA analysis.
6
Measuring Inflammatory Cytokines in Mouse and Human Cells
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ELISAs were performed as described previously16 (link). Using heparinized syringes, peripheral blood was obtained via the vena cava from mice. To dissociate femoral muscle, isolated muscle was cut into small pieces in PBS and dissociated in a gentleMACS C tube using a gentle MACS dissociator (Miltenyi Biotec, Bergisch Gladbach, Germany). Measurement of G-CSF, IL-6, IL-1β, and TNF-α levels was performed using mouse Quantikine ELISA kits in accordance with the manufacturer’s instructions (R&D Systems, MN, USA).
HUVECs were purchased from PromoCell, and the cells were cultured in Endothelial Cell Growth Medium 2 (PromoCell) in accordance with the manufacturer’s protocols. After treatment of the cells with α-toxin and PGN, the culture supernatants were harvested, and the G-CSF levels were measured using a human G-CSF Quantikine ELISA Kit (R&D Systems, MN, USA).
HUVECs were purchased from PromoCell, and the cells were cultured in Endothelial Cell Growth Medium 2 (PromoCell) in accordance with the manufacturer’s protocols. After treatment of the cells with α-toxin and PGN, the culture supernatants were harvested, and the G-CSF levels were measured using a human G-CSF Quantikine ELISA Kit (R&D Systems, MN, USA).
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