The spleens were collected and processed into single‐cell suspensions. NK cells were isolated from splenocytes to >90% purity, starting with cells from two pooled spleens, using the EasySep Mouse NK Cell Isolation Kit (Stemcell Technologies, Vancouver, CA) following the manufacturer's instructions. Target YAC‐1 cells were labeled with 100 μCi of Chromium‐51 (51Cr; Perkin Elmer, Waltham, MA, USA) for 75 min. A total of 10 000 labeled target cells were added to the wells of V‐bottomed plates, followed by effector NK cells at various effector‐to‐target ratios. Spontaneous 51Cr release was determined in YAC‐1 cultures (six wells) incubated in the medium alone. Hydrochloric acid (1 M) was added to three wells to maximize 51Cr release. After 4 h of incubation at 37°C and 5% CO2, supernatants were collected and 51Cr release was measured on the Wallac Wizard gamma Counter (Perkin Elmer, Waltham, MA, USA). We determined the percentage of labeled target cells that were lysed specifically by NK cells with the following formula:
Wallac wizard gamma counter
Manufactured by PerkinElmer
Sourced in United States
The Wallac Wizard Gamma Counter is a high-performance instrument designed for the quantification and analysis of gamma-emitting radioactive samples. It utilizes a sophisticated detection system to accurately measure the radioactivity levels in various types of samples, making it a versatile tool for a wide range of applications in research and clinical settings.
Automatically generated - may contain errors
6 protocols using wallac wizard gamma counter
1
NK Cell-Mediated Cytotoxicity Assay
2
PET Imaging of Tumor Response to RXC004
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experiment was performed at Medicines Discovery Catapult. SNU-1411 (1 × 107 cells; NOD-SCID mice) or HPAF-II (1 × 107 cells; NOD-SCID mice) were implanted subcutaneously and treatment with RXC004 (5 mg/kg once daily) or vehicle commenced when tumors reached approximately 200 mm3. Mice were imaged prior to dosing, day 3 and day 7 after dosing. A 10 MBq injection of 18FDG (PETNET), with a 45-minute washout was followed by a 20-minute PET scan using a Siemens Inveon PET scanner. Tissues removed on day 7 for biodistribution analysis were assessed using a PerkinElmer Wallac Wizard Gamma counter.
3
Carbohydrate-Binding Protein Affinity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plates of removable polystyrene wells coated with streptavidin (Thermo Scientific Pierce) were washed 3× with binding buffer (150 mM NaCl, 25 mM Tris–Cl, pH 7.8, 2.5 mM CaCl2), incubated for 2 h at 4°C with biotin-tagged CRDs at 5 μg/mL in 0.1% (w/v) BSA in binding buffer and again washed 3× with binding buffer. Serial 3-fold dilutions of ligands were made and buffer and reporter ligand were added so that the final concentrations corresponded to 0.5 μg/mL 125I-Man31-BSA in 0.1% BSA in binding buffer. After incubation for 2 h at 4°C, wells were washed 3× with binding buffer and counted in a Wallac WIZARD gamma counter (PerkinElmer Life Sciences). Inhibition constants were obtained by fitting to a simple binding curve (Stambach and Taylor 2003 (link)) using SigmaPlot. Ratios of different competing ligands were calculated for assays performed together on one assay plate. Values reported are means ± SD for 3–4 replicate experiments.
4
Biodistribution of 18F-CP18 in Mice
5
Biodistribution of 18F-Flotegatide in Atherosclerosis
6
Cytotoxicity Assay for Antibody-Mediated Cell Killing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Target cells (1 × 106) were labeled in their culture medium for 90 min at 37 °C with 100 µCi 51Cr (PerkinElmer, Waltham, MA, USA) and finally diluted to 0.1 × 106 cells/mL after several washing steps. Neutrophils were either left untreated or were pre-incubated with 10 µg/mL anti-SIRPα at room temperature for the indicated conditions. Co-incubation of target and effector cells was carried out at a target:effector (T:E) ratio of 1:50 (i.e., 5000:250,000 cells), unless specified otherwise, for 4 h at 37 °C and 5% CO2 in the absence or presence of 0.5 µg/mL dinutuximab. Spontaneous and maximum 51Cr release were determined by incubating the target cells without effector cells and by treating them with a 0.1% triton X-100 (Sigma Aldrich, St. Louis, MO, USA), respectively. After incubation, supernatant was harvested and analyzed for radioactivity in a Wallac Wizard gamma counter or a MicroBeta2 plate reader (PerkinElmer). The percentage of cytotoxicity was calculated as: [(experimental counts per minute (CPM)-spontaneous CPM)/(maximum CPM-spontaneous CPM)] × 100%. All conditions were performed in duplo or triplicate.
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!