ELISpot was performed as previously described [35 (link)] and according to the manufacturer’s instructions (R&D Systems, SEL485 and SEL404, Minneapolis, MN, USA). Briefly, rPfCelTOS (10 µg/mL) was used as the stimulating antigen. Hamster anti-mouse CD3e (1 µg/mL [BD Biosciences, 553057, San Jose, CA, USA]) was used as a positive control for cell stimulation. The negative control was culture media in place of a stimulating antigen. Splenocytes were plated at 2 × 105 cell/well. Plates were incubated for 48 h at 37 °C with 5% carbon dioxide. Spot counting was performed using an AID ELISpot Reader (Autoimmun Diagnostika, Strassberg, Germany).
Sel404
Manufactured by R&D Systems
Sourced in United States
The SEL404 is a laboratory equipment product. It is designed for performing specific tasks in a research or analytical setting. The core function of the SEL404 is to enable users to carry out essential procedures or analyses required for their work. Further details about the intended use or specific capabilities of the SEL404 are not available.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sel404
1
ELISpot Assay for PfCelTOS Antigen
2
Quantifying hFIX-Specific Cytokine Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ELISpot assays were performed for hFIX-specific IL-4 and IFN-γ responses using mouse IL-4 (SEL404) and IFN-γ development module (SEL485) according to manufacturer's protocol (R&D system, Minneapolis, MN, USA). Splenocytes were isolated from primed BALB/c and C3H/HeJ haemophilia B mice. 106 splenocytes were cultured in 200 μL of RPMI 1640 with 10% FBS, 1% penicillin/streptomycin, 15 mM Hepes (pH7.2) and 55 μM 2-beta-mercaptoethanol, with or without the stimulation of 10 μg mL−1 hFIX protein for 14 to 16 h (IFN-γ) or 48 h (IL-4) at 37°C in a 5% CO2 incubator. Staphylococcal Enterotoxin B (1 μg 100 μL−1; Sigma-Aldrich, St. Louis, MO, USA), and PMA-Phorbol 12-myristate 13-acetate (0.05 g mL−1)/Ionomycin (1 μg mL−1; Sigma-Aldrich), were used as positive controls. Spots were analysed and counted with the CTL-ImmunoSpotH S5 UV analyser (Cellular Technology, Shaker Heights, OH, 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!