Snap‐frozen livers from three animals exposed to each condition were pooled and homogenized in lysis buffer, as described previously 16 . Lysates were analyzed according to the manufacturer's instructions, with a Human Cytokine Array C3 (RayBiotech, Norcross, GA, USA).
Human cytokine array c3
Manufactured by RayBiotech
Sourced in United States
The Human Cytokine Array C3 is a multiplex assay that can simultaneously detect the relative levels of 80 different human cytokines, chemokines, and soluble receptors in a single sample. The array is designed to provide a comprehensive analysis of the inflammatory response or immune status of a biological sample.
Automatically generated - may contain errors
Lab products found in correlation
Il 8 elisa kit, by R&D Systems (2 mentions)
Il 6 and, by R&D Systems (1 mentions)
Imagequant las 4000 system, by GE Healthcare (1 mentions)
Imagequant tl, by GE Healthcare (1 mentions)
Il 6 human uncoated elisa kit, by Thermo Fisher Scientific (1 mentions)
Elisa, by BD (1 mentions)
Elisa kit, by R&D Systems (1 mentions)
Image studio lite, by LI COR (1 mentions)
Las 4000 imager, by Fujifilm (1 mentions)
8 protocols using human cytokine array c3
1
Multiplexed Cytokine Analysis in Liver
2
Cytokine Profiling of Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokine antibody arrays (“Human Cytokine Array C3”, Raybiotech, Norcross, GA, USA) were performed after 7, 14, and 21 days of culture according to the manufacturer’s instructions. Briefly, 10-fold diluted culture supernatants coming from 3 independent donors or medium alone were incubated on antibody-coated membranes before detection with a streptavidin-horseradish peroxidase biotinylated-antibody complex and chemiluminescence detection on X-ray films for 2 min. The integrated optical density (IOD) of each dot was measured on digitalized autoradiograms with the ImageJ software (1.50i, National Institutes of Health, Bethesda, MD, USA) after subtraction of the non-specific signal. Each value corresponds to the ratio between the dot studied and the internal positive control (expressed as the average of six internal control dots). Values under the detection threshold (determined by the maximum value of the negative internal control dots) were considered as non-achievable (NA). Data were calculated as the average level of secretion (arbitrary units) for each mediator normalized with internal positive controls and internal negative control.
3
Profiling Cytokine Secretion in PBMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PBMCs were isolated from buffy coat preps and either induced or left un-induced as described above. Both induced and un-induced cells were maintained at 37°C in a humidified atmosphere with 5% CO2. At 48hrs post-induction, growth media from induced or un-induced cells were collected, centrifuged at 3000 rpm for 10 min to remove the cellular debris. Cytokine/chemokine profiles of conditioned media were determined by using a commercially available kit (RayBiotech INc., human cytokine array C3, code: AAH-CYT-3) according to the manufacturer's instructions.
4
Quantifying UVB-Induced Cytokine Profiles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Media from NHEKs stimulated with UVB radiation were harvested and centrifuged for 15 min at 4 °C. The supernatants were freeze-dried and used for multiple cytokine measurements with a human cytokine array C3 (Raybiotech, GA, USA), according to the manufacturer’s instructions. The IL-6 and IL-8 levels were quantified using IL-6 and IL-8 ELISA kits, respectively, according to the manufacturer’s instructions (R&D Systems).
5
Cytokine Profiling of Tumor-Associated Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokines from resting and TAM-like macrophages were analyzed by Human Cytokine Array C3 (Ray Biotech, GA, USA). Signals were measured using an ImageQuant LAS 4000 system and ImageQuant TL software (GE Healthcare, Uppsala, Sweden). The signal from an individual spot on a membrane exposed to TAM-like-CM was normalized by positive controls prior to normalization with corresponding spots on resting macrophage membrane.
6
Cytokine Quantification in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human cytokines were measured using human IL-6 uncoated ELISA kit (Invitrogen, ThermoFisher Scientific, UK), human TNF standard ABTS ELISA kit (Peprotech, London, UK), and human IL-8 ELISA MAX standard kit (Biolegend, San Diego, CA, USA) according to the manufacturers’ instructions. Cytokine arrays used were Human Cytokine Array C3 (RayBiotech, Norcross, GA, USA). The levels of mouse cytokines present in culture supernatants were quantified using an ELISA (BD Pharmingen, North Ryde, NSW, Australia), according to the manufacturer’s instructions.
7
Secretome Profiling of Activated Bone Marrow Mesenchymal Stem Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
aBMSCs and BMSCs were cultured to 75–80% confluent in T-25 flasks and then washed briefly with DPBS and incubated with 2.5 mL basal medium (MEM α without serum) for 24 h. The conditioned media (CM) were collected, centrifuged at 4 °C to remove cellular debris, and stored at − 80 °C until use. The secretion of cytokines and growth factors by aBMSCs and BMSCs was assessed by a Human Cytokine Array C3 (RayBiotech, Peachtree Corners, GA, USA; detecting 42 human cytokines: ENA-78, GCSF, GM-CSF, GRO, GRO-α, I-309, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 p40/p70, IL-13, IL-15, IFN-γ, MCP-1, MCP-2, MCP-3, MCSF, MDC, MIG, MIP-1δ, RANTES, SCF, SDF-1, TARC, TGF-β1, TNF-α, TNF-β, EGF, IGF-I, Angiogenin, Oncostatin M, Thrombopoietin, VEGF-A, PDGF BB, Leptin). The concentrations of IL-6 and MCP-1 in CM were evaluated by ELISA kits (R&D Systems, Minneapolis, MN, USA).
8
Profiling Cytokine Secretion in UVB-Stressed NHEKs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The medium was harvested from NHEKs, stimulated with UVB radiation, and centrifuged for 15 min at 4 °C. The supernatants were freeze-dried and used for multiple cytokine measurements with Human Cytokine Array C3 and Human Cytokine Array C4 Kits (RayBiotech, Norcross, GA, USA) according to the manufacturer’s instructions. The arrays were imaged with a Fujifilm LAS-4000 imager. Image analysis was performed using LI-COR Image Studio Lite. Each dot was assigned a value, and the average of two measurements was calculated for each array. The average density values for each cytokine in each condition were then averaged for each array to calculate the fold change. IL-6, IL-7, and IL-8 levels were quantified using IL-6, IL-7, and IL-8 ELISA kits, respectively, according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!