Ben-Men-I cells were seeded at a concentration of 6 × 104/mL in DMEM medium and incubated with unlabeled apoptotic cells. After 24 h, the culture supernatants were harvested, centrifuged at 1,000 rpm for 5 min, and assayed for IL-6, IL-8 (Orgenium Laboratories), IL-1RA (R&D, DRA00B), IL-16 (RayBiotech, ELH-IL16-001), MIF (RayBiotech, ELH-MIF-001), or CXCL1 (R&D, DRG00) by ELISA. Cytokines present in apoptotic body preparations were measured and background subtracted. The assays were performed according to the manufacturers’ instructions. Absorbance was read on a Spectramax GEMINI XS microplate reader (450 nm). For multiplexed detection of cytokines/chemokines, the human cytokine array panel A (R&D, ARY005) was used according to manufacturers’ recommendations.
Drg00
Manufactured by R&D Systems
Sourced in United States
DRG00 is a lab equipment product. It is designed for general laboratory use. The core function of DRG00 is to perform basic laboratory tasks.
Automatically generated - may contain errors
Lab products found in correlation
Ary005, by R&D Systems (1 mentions)
Ab46027, by Abcam (1 mentions)
Ab173194, by Abcam (1 mentions)
Ab46048, by Abcam (1 mentions)
St51011, by R&D Systems (1 mentions)
Ab214570, by Abcam (1 mentions)
Mak263, by Merck Group (1 mentions)
Quantikine elisa kit, by R&D Systems (1 mentions)
Cobas 8000, by Roche (1 mentions)
Il 10, by BD (1 mentions)
Interleukin 4 (il 4), by BD (1 mentions)
Dugb00, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by BD (1 mentions)
Interleukin 2 (il 2), by BD (1 mentions)
6 protocols using drg00
1
Cytokine Profiling of Apoptotic Cell-Stimulated Macrophages
2
Serum Biomarker Profiling via ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum concentrations of interleukin (IL) 6 (Abcam, United States, ab46027), interferon (IFN) γ (Abcam, United States, ab46048), interferon-inducible protein (IP) 10 (Abcam, United States, ab173194), interferon α 1 (Abcam, United States, ab213479), IL-10 (Abcamn United States, ab100549), and HMGB-1 (IBL, Germany, ST51011), and the serum receptor for advanced glycation end-product (sRAGE, R&D, United States, DRG00) were detected via commercial ELISA kits following the instructions provided by the manufacturers (detailed in Supplementary materials ).
3
Quantifying RAGE, LGALS3BP, and PTX3
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
4
Quantifying Plasma Biomarkers in Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total plasma sRAGE was measured via enzyme-linked immunosorbent assay (ELISA; DRG00, R&D Systems, Minneapolis, MN, USA). This total sRAGE quantification approach included the cleaved (cRAGE) and endogenous secretory (esRAGE) isoforms. To quantify plasma esRAGE specifically, another ELISA was utilized (K1009-1AS, One International, Mountain View, CA, USA). Plasma cRAGE was calculated by subtracting esRAGE from total sRAGE [33 (link),34 (link),35 (link)]. Proinflammatory cytokines (IL-6, IL-1β, and TNFα) were analyzed in duplicate by 5-plex electrochemiluminescence immunoassay (MSD, Rockville, MD, USA). IL-1Ra was measured using a Quantikine ELISA Kit (R&D Systems). Urinary creatinine was measured via absorption photometry (Cobas 8000, Roche Diagnostics, Basel, Switzerland), and free 8-iso prostaglandin F2α (8-iso PGF2α), a marker for oxidative stress [36 (link)], was assessed in triplicate using an 8-isoprostane ELISA (Cayman Chemicals, Ann Arbor, MI, USA). To evaluate urine glucose concentration [37 ], 24-h urine collections were assessed via the glucose oxidase colorimetric method (MAK263, Sigma-Aldrich, St. Louis, MO, USA).
5
Cytokine and Lung Marker Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum samples were diluted 1:5 times for ELISA analysis. Standards were prepared according to the manufacturer’s protocol. ELISA kits were used to prepare the samples according to each antibody. Cytokines analyzed include IL2 (0017212, BD Biosciences), IL6 (550799, BD Biosciences), IL8 (550999, BD Biosciences), IL4 (550614, BD Biosciences), TNF-α (550610, BD Biosciences), and IL10 (550613, BD Biosciences) and Lung markers include Uteroglobin (DUGB00, R&D Systems), SP-D (DSFPD0, R&D Systems), and sRAGE (DRG00, R&D Systems). Samples were processed in a BSL2 cabinet with appropriate PPEs.
6
Quantification of Soluble RAGE Isoforms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma total sRAGE concentrations were measured using a commercially available ELISA (R&D ELISA, DRG00). The AsOne International ELISA (K-1009-1) measures only esRAGE. From this, we calculated cell surface cleaved (c) sRAGE (total sRAGE – esRAGE) = cRAGE. For total sRAGE, each unique sample was analyzed in duplicate (50 µL/replicate). The sensitivity of the assay was 16.14 pg/mL, with an assay range of 78–5000 pg/mL. There was <0.5% cross-reactivity with other molecules. The intra-assay precision (CV) was ∼5.7% and the interassay precision (CV) was ∼7.7%. For esRAGE, each unique sample was analyzed in duplicate (20 µL/replicate). The linear range of the esRAGE measurements was 0.05–3.2 ng/mL. The inter- and intra-assay CVs for esRAGE were <10% and <4.4%, respectively. For every experiment, standards were included as per the manufacturer's instructions.
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!