Mice were dosed with 7.5 μg of luciferase mRNA complexed with bAC CART or LNP. Blood samples were collected from the mice 24 h after treatment. Around 500 μl of blood per sample were submitted to the Stanford Diagnostics Lab for hematological analysis. Assessed parameters included Complete Blood Count (CBC) with white blood cell differential, electrolytes, liver function tests, kidney function tests, and glucose. Blood samples collected 24 h after CART/mRNA injection were also processed to isolate serum for Luminex analysis conducted by the Human Immune Monitoring Center at Stanford University. This analysis was performed using the Mouse 48 plex Procarta kit (Thermo Fisher/Life Technologies) following manufacturer instructions.
Mouse 48 plex procarta kit
Manufactured by Thermo Fisher Scientific
The Mouse 48-plex Procarta kits are multiplexed assays designed to measure the levels of 48 different mouse protein analytes in a single sample. The kits utilize Luminex xMAP technology to enable the simultaneous quantification of multiple targets.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using mouse 48 plex procarta kit
1
Comprehensive Evaluation of Systemic Response to mRNA Therapy
2
Multiplex Cytokine Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This assay was performed by the Human Immune Monitoring Center at Stanford University as previously described (47 (link)). Mouse 48 plex Procarta kit (Thermo-Fisher/Life Technologies) was used according to the manufacturer's instructions but with some modifications. The samples were added to a plate containing beads that were linked with antibodies and incubated overnight at 4°C with shaking. Then, a biotinylated detection antibody was added for 60 min at room temperature with shaking. After washing the plate, streptavidin-PE was added for 30 min at room temperature, and the plate was washed again. Reading buffer was added to the wells, and each sample was measured in duplicate. The plates were read using a Luminex 200 or a FM3D FlexMap instrument with a lower bound of 50 beads per sample per cytokine. Custom Assay Chex control beads were added to all wells.
3
Multiplex Cytokine Assay for Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assay was performed by the Human Immune Monitoring Center at Stanford University. Mouse 48-plex Procarta kits (EPX480-20834-901) were purchased from Thermo Fisher and used according to the manufacturer’s recommendations with modifications as described below. Beads were added to a 96-well plate and washed in a BioTek ELx405 washer. Samples were added to the plate containing the mixed antibody-linked beads and incubated overnight at 4 °C with shaking. Cold (4 °C) and room temperature incubation steps were performed on an orbital shaker at 500–600 rpm. Following the overnight incubation, plates were washed in a BioTek ELx405 washer and biotinylated detection antibody was added for 60 min at room temperature with shaking. Plates were washed as described above and streptavidin-PE was added. After incubation for 30 min at room temperature, a wash was performed as above and reading buffer was added to the wells. Each sample was measured in singlets. Plates were read on a FM3D FlexMap instrument with a lower bound of 50 beads per sample per cytokine/chemokine. Custom Assay Chex control beads were purchased from Radix BioSolutions, and were added to all wells.
4
Cytokine Analysis of Inner Ear Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The inner ears (n = 3) were dissected out and the surrounding soft tissues were removed at 7 days. Inner ears were washed with fresh PBS > 20 times to clean the surface of the cochleae. Each wash procedure was performed carefully in sterilized dishes, which were changed at each wash step. Samples were homogenized mechanically in lysis buffer. This buffer included 1% Triton-X 100 (9002–93-, Sigma-Aldrich, St. Louis, MO, USA), 0.5% NP-40 (FNN0021, Thermo Fisher Scientific, Waltham, MA, USA), 25 mM Tris–HCl pH 7.5 (1185-53-1, Millipore Sigma, Burlington, MA, USA), 100 mM NaCl (764714-5, Sigma-Aldrich), Halt protease inhibitor cocktail (78430, Thermo Fisher Scientific) and phenylmethanesulfonyl fluoride (32998-6, Millipore Sigma). Samples were stored at − 80 °C and cytokine analysis was performed at the Human Immune Monitoring Core (Stanford University) as previous described [16 (link)]. Briefly, Mouse 48-plex Procarta kits (EPX480-20834901, Thermo Fisher Scientific) were employed and plates were read using FM3D FlexMap instrument with a lower bound of 50 beads per sample per cytokine. Custom Assay Chex control beads were added to all samples. Each sample was tested in triplicate. MFI was averaged over duplicate wells for each cytokine per sample on each plate.
5
Multiplex Cytokine Assay for Mice
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!