To evaluate epitope-specific T cell responses, splenocytes were collected from vaccinated mice 7 days after their last immunizations. After splenocytes were purified with lympholyte-M (CL5031, Cedarlane), 0.25 million splenocytes were plated in each well of a 96-well ELISPOT plate (Millipore, MSIPS4510), in 200 μl per well. The cells were either left unstimulated as negative control or stimulated with peptides (1 × 10−3 mM Trp2, or 1 × 10−3 mM Td) or ConA (positive control, C5275, Sigma) for 20 hours at 37°C in a humidified CO2 incubator. IFNγ ELISPOT pairs (551881, BD), streptavidin–alkaline phosphatase (3310-10-1000, Mabtech), and Sigmafast BCIP/NBT substrate (B5655, Sigma) were used to detect and develop antigen-specific spots. Plates were enumerated by ZellNet Consulting using a Zeiss KS ELISPOT system.
Ks elispot system
Manufactured by Zeiss
The KS ELISPOT system is a laboratory equipment used for the detection and enumeration of antigen-specific cells that secrete cytokines or other soluble proteins. It utilizes the ELISPOT (Enzyme-Linked ImmunoSpot) technique, which allows for the sensitive and quantitative analysis of individual cells producing a specific protein of interest. The core function of the KS ELISPOT system is to facilitate the accurate and reproducible measurement of immune responses in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin alkaline phosphatase, by Mabtech (4 mentions)
Msips4510, by Merck Group (4 mentions)
Sigmafast bcip nbt substrate, by Merck Group (2 mentions)
Ifnγ elispot pairs, by BD (2 mentions)
Cas9 protein, by Mabtech (2 mentions)
Mouse ifn γ elispot pairs 3321 3 250, by Mabtech (2 mentions)
Sigmafast bcip nbt, by Merck Group (2 mentions)
Cas9 protein, by Applied Biological Materials (2 mentions)
Cl5031, by Cedarlane (1 mentions)
C5275, by Merck Group (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
Ack lysis buffer, by Thermo Fisher Scientific (1 mentions)
Lympholyte m, by Cedarlane (1 mentions)
4 protocols using ks elispot system
1
Evaluating Epitope-Specific T Cell Responses in Vaccinated Mice
2
Quantifying T-cell Responses to Cas9 Protein
3
Quantifying T-cell Responses to Cas9 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
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T-cell ELISPOTS were performed as previously described [44 (link)]. Briefly, splenocytes from AAV9-SaCas9-injected mice were isolated and purified using Lympholyte M (Cederlane). 250,000 cells were mixed with either Cas9 protein (Applied Biological Materials) or cell media only as a negative control, and subsequently plated in a 96-well ELISPOT plate (Millipore, MSIPS4510) in 100 µL per well. Stimulation was performed with 0.02 µg/µL of Cas9 protein at 37ºC in a humidified incubator, 7% (v/v) CO2, for 40 h. Mouse IFN-γ ELISPOT pairs (3321–3-250 and 3321–6-250) and Streptavidin-alkaline phosphatase (3310–10) were purchased from Mabtech. Spots were developed using substrate Sigmafast BCIP/NBT (Sigma, B5655). Plates were shipped to Zellnet Consulting and enumerated using a Zeiss KS ELISPOT system. Full plates images are reported in Extended Data Fig. 4 .
4
Quantifying Antigen-Specific Immune Responses
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