BV-2 cells were treated for 24 h and then supernatants from controls (dH2O only), from cells treated with GLE (0.5 mg/ml) only, LPS only (1.0 μg/ml), and GLE (0.5 mg/ml) + LPS (1.0 μg/ml) (where LPS was added after 1 h incubation with GLE), were used in the assay. Specific ELISAs (RayBiotech, Norcross, GA, USA) were performed using G-CSF (Cat# ELM-G-CSF), IL1α (Cat# ELM-IL1a), MCP-5 (Cat# ELM-MCP5), MIP3α (Cat# ELM-MIP3a), and RANTES (Cat# ELM-RANTES) following manufacturer’s instructions and as previously described (Mendonca et al., 2017 (link)). Briefly, 100 μl of supernatant from samples and standards were added to 96 well plates pre-coated with the capture antibody. After incubation, 100 μl of prepared biotinylated antibody mixture was added to each well and incubated for 1 h. The mixture was then decanted, and streptavidin solution (100 μl) was placed in each well and incubated. Substrate reagent (100 μl) was then added to each well for 30 min, followed by the addition of stop solution (50 μl). Data were quantified by optical density at 450 nm.
G csf
Manufactured by RayBiotech
Sourced in United States
G-CSF is a recombinant granulocyte colony-stimulating factor. It is a protein that stimulates the production and release of neutrophils, a type of white blood cell, from the bone marrow.
Automatically generated - may contain errors
Lab products found in correlation
Rantes, by RayBiotech (2 mentions)
Specific elisas, by RayBiotech (2 mentions)
Mcp 5, by RayBiotech (2 mentions)
Mip3α, by RayBiotech (2 mentions)
Mip3a, by RayBiotech (2 mentions)
Gm csf, by RayBiotech (2 mentions)
Basic fibroblast growth factor (bfgf), by Cusabio (1 mentions)
Angii, by RayBiotech (1 mentions)
Il 10, by RayBiotech (1 mentions)
4 protocols using g csf
1
ELISA Analysis of Inflammatory Factors in BV-2 Cells
2
ELISA Analysis of Inflammatory Factors in BV-2 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BV-2 cells were treated for 24 h and then supernatants from controls (dH2O only), from cells treated with GLE (0.5 mg/ml) only, LPS only (1.0 μg/ml), and GLE (0.5 mg/ml) + LPS (1.0 μg/ml) (where LPS was added after 1 h incubation with GLE), were used in the assay. Specific ELISAs (RayBiotech, Norcross, GA, USA) were performed using G-CSF (Cat# ELM-G-CSF), IL1α (Cat# ELM-IL1a), MCP-5 (Cat# ELM-MCP5), MIP3α (Cat# ELM-MIP3a), and RANTES (Cat# ELM-RANTES) following manufacturer’s instructions and as previously described (Mendonca et al., 2017 (link)). Briefly, 100 μl of supernatant from samples and standards were added to 96 well plates pre-coated with the capture antibody. After incubation, 100 μl of prepared biotinylated antibody mixture was added to each well and incubated for 1 h. The mixture was then decanted, and streptavidin solution (100 μl) was placed in each well and incubated. Substrate reagent (100 μl) was then added to each well for 30 min, followed by the addition of stop solution (50 μl). Data were quantified by optical density at 450 nm.
3
Quantification of Secreted Factors in Tumor Microenvironment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Snap-frozen tumor samples were crushed into fragments <2 mm in diameter and resuspended with 1:2 weight/volume of 2×PBS and rotated at 4°C for 2 h. The samples were vortexed then spun for 3 min at 5,000 RPM then the tumor extracellular fluid was collected36 (link). Collected cultured cells and diluted to 2 × 106/ml with PBS. Broke up cells by ultrasonic and spun for 10 min at 10000 g then the cell lysate was collected. Serum samples and/or tumor extracellular fluid and/or cell lysate from the control and SH-treated groups were applied to ELISA kits, following the manufacturer 's protocol to quantify bFGF (CUSABIO, China), PF-4 (CUSABIO, China), G-CSF (RayBiotech, USA) and GM-CSF (RayBiotech, USA).
4
Quantification of Signaling Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AngII, IL-10, GM-CSF, G-CSF, Eotaxin-2, and CXCL11 ELISA kits were all obtained from Ray Biotech, Inc. (Norcross, GA, USA). TNFSF14 ELISA kits was obtained from CUSABIO Inc. (Wuhan, China). The ELISA for them was carried out according to the manufacturer’s instructions. Two independent experiments were performed, each in triplicate. For AngII ELISA, the microplate in the kit, which is pre-coated with anti-rabbit secondary antibody, was incubated with an anti-AngII antibody, under such conditions that both biotinylated AngII peptide and a peptide standard or targeted peptide in the samples interacted competitively with the AngII antibody. Unbound biotinylated AngII peptide was then allowed to interact with streptavidin-horseradish peroxidase (SA-HRP), which catalyzes a color development reaction. The intensity of the colorimetric signal is directly proportional to the amount of the biotinylated peptide-SA-HRP complex and inversely proportional to the amount of the AngII peptide in the standard or samples. A standard curve of known concentration of AngII peptide was established, and the concentration of AngII peptide in the samples was then calculated by interpolation onto the standard curve. All groups of tumor cells were all seeded at a density of 200,000 cells/well in 1 ml medium in 24-well plates.
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!