One million MC38 cells were implanted subcutaneously in the right hind flank of mice. Tumor growth was monitored using electronic calipers and calculated according to the formula: V = length × width2 × 0.52. For T cell phenotypic analysis by flow cytometry, spleens and inguinal lymph nodes were harvested 17 days post-treatment initiation. Splenic and tdLN cells were stained with anti-mouse antibodies for flow cytometry analysis. To deplete T cells, mice received intraperitoneal injection of 200 μg anti-CD4 (BioXcell BE0003) and 200 μg anti-CD8 (BioXcell BE0061) antibodies in PBS, a second dose of the antibodies was administered two days later. To asses cytotoxicity, MC38 cells were thawed, seeded at a density of 3 × 104 in a flat bottom 96-well plate and treated overnight at 37°C and 5% CO2 with AZD-7762 (MCE HY-10992) at the indicated concentrations. Cell viability was measured with PrestoBlue (Invitrogen).
Be0003
Manufactured by BioXCell
The BE0003 is a centrifuge designed for general laboratory applications. It features a maximum speed of 6,000 rpm and a maximum relative centrifugal force of 4,500 x g. The centrifuge can accommodate rotors with a capacity of up to 4 x 100 mL.
Automatically generated - may contain errors
4 protocols using be0003
1
Subcutaneous Tumor Growth Monitoring and T Cell Analysis
2
Tumor Models and Immune Depletion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For tumor models, 2 × 105 tumor cells were injected s.c. unless otherwise specified. Tumor growth was monitored daily and measured every 3–4 days using digital callipers. Tumor volume was calculated by multiplying the longest tumor diameter and the square of its perpendicular diameter. For depletion experiments, mice were injected intraperitoneally with 100 μg isotype control (Bio X cell; BE0085 and BE0088), anti-NK1.1 (Bio X cell; BE0036), anti-CD4 (Bio X cell; BE0003) or anti-CD8β (Bio X cell; BE0223) at indicated days. shRNA sequences used to inhibit mouse CD8+ T Rig-I expression were GTTAACCCACAGTTGATCCAAATGATACTCGAGTATCATTTGGATCAACTGTGGTTTTTTCTCGAG (Rig-I-sh1) and GTTAACCAAGCATTCAGAGACTATATCCTCGAGGATATAGTCTCTGAATGCTTGTTTTTTCTCGAG (Rig-I-sh2). For STAT5 inhibitor treatment, mice were given 3 doses of i.p. injection of SH-4-54 (10 mg/kg) or STAT5-IN-1 (20 mg/kg) or vehicle control every 3 days from when the tumor was visible.
3
Immune Cell Depletion Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Depletion antibodies for CD4+ (BE0003), CD8+ (BE0061), or NK1.1 (BE0036) were obtained from BioXCell. Mice were depleted prior to starting the experiment via four once-daily administrations of 100 µg depletion antibodies given intraperitoneally. This depletion was maintained throughout the study by twice weekly administrations of 100 µg of antibody. The depletion protocol was verified via flow cytometry (Fig. S1).
4
Modulating Tumor Immunity with Antibodies
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!