Cytokine levels produced by four iPSC‐derived NK cells were detected using a human IL24 (Abclonal, #RK00112), GM‐CSF (Abclonal, # RK04505), TNF‐α, and IFN‐γ ELISA kit (Invitrogen #KHC4021, #BMS2034) according to manufacturer instructions. A total of 1 × 106 iNK cells were seeded with the stimulus of PMA (TopScience; #TQ0198) and ionomycin (TopScience; #T7285). Abiding by the incubation of 37°C, 24 h, cell‐free culture supernatants were used for detection. ELISA was performed with two technical replicates for all samples. Measurements were performed in triplicates.
Khc4021
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The KHC4021 is a compact centrifuge designed for general laboratory use. It features a maximum speed of 4,000 rpm and can accommodate a rotor with a capacity of up to 12 x 1.5/2.0 mL microtubes. The centrifuge is operated through a simple control panel and has a timer function.
Automatically generated - may contain errors
Lab products found in correlation
Bms2221hs, by Thermo Fisher Scientific (2 mentions)
Prestoblue cell viability reagent, by Thermo Fisher Scientific (2 mentions)
Ek0525, by Boster Bio (1 mentions)
K113 100, by R&D Systems (1 mentions)
Anti his tag antibody, by Thermo Fisher Scientific (1 mentions)
Celltiter glo, by Promega (1 mentions)
6 protocols using khc4021
1
Quantification of Cytokine Secretion in iPSC-NK Cells
2
ELISA for Quantifying IFN-γ
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Release of IFN γ was assayed by a sandwich ELISA Kit (KHC4021, Invitrogen Corp., Camarillo, CA, USA). The detection of IFN-γ in cell culture supernatant was performed according to the manufacturer instructions. Human IFN-γ was diluted (1000–6 pg/mL) and used as a standard to establish the standard curve.
3
Cytotoxic T Cell-Mediated Cancer Killing
4
Quantification of Serum Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5
Cytotoxic T Cell-Mediated Cancer Killing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2.5 × 104 synNotch CD4+ or CD8+ primary T cells or murine cell lines were co-cultured with target cancer cells at a 1:1 ratio, with the addition of ICEp at 0.075 OD550 × 200 μL medium (100 μL T cell medium + 100 μL cancer cell medium) (unless otherwise specified). Dual antigen-positive (GFP and HER2) target cells (A375 and K562) and GFP-positive K562 cells were used as positive controls. After mixing, cells were centrifuged for 2 minutes at 300 x g to initiate interaction between the cells. After 24 hours, the co-culture from CD8+ T cells were stained with anti-CD69 antibody (BD #562884), analyzed by flow cytometry (BD LSR II), and imaged through the spinning disk confocal microscope (Nikon Yokogawa CSU-22). After 48 hours, cytokine concentration in the supernatant was measured by IL-2 ELISA (for CD4+ T cells, eBiosciences #BMS2221HS) and Interferon-γ (IFN-γ) ELISA (for CD8+ T cells, ThermoFisher #KHC4021).
To quantify target cell killing, 2.5 × 104 A375 cells were seeded on a flat-bottom 96-well tissue culture plate (Falcon #353072) and cultured for 8 hours, followed by the addition of 2.5 × 104 CD8+ T cells and ICEp at 0.075 OD550 × 200 μL final (unless otherwise specified). After 1–3 days, cells were gently washed with PBS twice and analyzed for cell viability using PrestoBlue Cell Viability Reagent (Invitrogen #A13262).
To quantify target cell killing, 2.5 × 104 A375 cells were seeded on a flat-bottom 96-well tissue culture plate (Falcon #353072) and cultured for 8 hours, followed by the addition of 2.5 × 104 CD8+ T cells and ICEp at 0.075 OD550 × 200 μL final (unless otherwise specified). After 1–3 days, cells were gently washed with PBS twice and analyzed for cell viability using PrestoBlue Cell Viability Reagent (Invitrogen #A13262).
6
Evaluating Cytokine-Induced Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To challenge cells with cytokines, cells were incubated with Interferon-γ (IFNγ; Cell Signaling Technology 80385S), FasL (AdipoGen AG-40B-0130-3010), TRAIL (R&D Systems 375-TL-010), or TNF-α (AdipoGen AG-40B-0019-3010) for 24 h. TRAIL was crosslinked by incubating with anti-His Tag antibody (Thermo Fisher Scientific MA121315, 1:500) for 15 min at room temperature. Cell viability was measured using CellTiter-Glo (Promega G7571) and protein was harvested for western blots. For evaluating Caspase 8 activity, cells were incubated with FasL or crosslinked TRAIL for 3 h and harvested for Caspase 8 colorimetric assay (R&D Systems K113-100). IFNγ in the cell culture media of the T cell cytotoxic assay was quantified using an ELISA kit (Thermo Fisher Scientific KHC4021).
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!