Flat-bottom 96-well plates (Corning) were coated overnight with goat anti-human Ig (1 mg/ml; Southern Biotech) at 4°C, washed with PBS/0.05%Tween-20, and blocked with PBS/5% FCS for 2 hours at room temperature. Samples were added for 1.5 hours. After washing, peroxidase-conjugated goat anti-human IgG (Thermo Fisher Scientific) or rabbit anti-human IgM (Jackson ImmunoResearch) was used to detect bound antibody. TMB Substrate (Thermo Fisher Scientific) was used to reveal peroxidase activity. Reactions were stopped with sulfuric acid and optical densities were measured at 450 nm using a BioTek Synergy 2 reader (Winooski). Concentrations were calculated using standard curves that were generated for each assay.
Rabbit anti human igm
Manufactured by Jackson ImmunoResearch
Sourced in United States
Rabbit anti-human IgM is a laboratory reagent used to detect and measure the presence of IgM antibodies in human samples. It is produced by immunizing rabbits with human IgM and purifying the resulting antibodies.
Automatically generated - may contain errors
Lab products found in correlation
96 well flat bottom plate, by Corning (2 mentions)
Synergy 2 reader, by Agilent Technologies (2 mentions)
Peroxidase conjugated goat anti human igg, by Thermo Fisher Scientific (2 mentions)
Goat anti human ig, by Southern Biotech (2 mentions)
Tmb substrate, by Thermo Fisher Scientific (1 mentions)
3 3 5 5 tetramethylbenzidine substrate, by Thermo Fisher Scientific (1 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Ibrutinib, by Selleck Chemicals (1 mentions)
Myc inhibitor 10058 f4, by Selleck Chemicals (1 mentions)
3 protocols using rabbit anti human igm
1
ELISA for Detecting Human Antibodies
2
Quantification of IgM and IgG in Cultured Memory B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IgM and IgG levels were determined in supernatants of memory B cells cultured for 6 days using ELISA. After overnight coating with goat anti-human Ig (1 mg/ml; Southern Biotech, Birmingham, USA) at 4 °C, flat-bottom 96-well plates (Corning, Tewksbury, USA) were washed with PBS/0.05% Tween-20 and subsequently blocked with PBS/5%FCS for 2 h at RT. Samples were added for 1.5 h at room temperature. After washing, peroxidase-conjugated goat anti-human IgG (Thermo Fisher Scientific) or rabbit anti-human IgM (Jackson, Uden, The Netherlands) were used to detect bound antibody. 3,3′,5,5′-Tetramethylbenzidine substrate (Thermo Fisher Scientific) was used to reveal peroxidase activity. Reactions were stopped with sulfuric acid and optical densities were measured at 450 nm using a BioTek Synergy 2 reader (Winooski, USA). Concentrations were calculated using standard curves for IgM and IgG.
3
BCR-Dependent and BCR-Non-Dependent DLBCL Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BCR-dependent DLBCL cell lines (TMD8, HBL1) and BCR-non-dependent/OxPhos-DLBCL cell lines (Toledo, LY4) [10, 15] were used in this study. TMD8 and HBL1 were kindly provided by Dr. Lynn Y. Wang (University of Chicago, Chicago, IL, USA). Toledo and LY4 were purchased from American Tissue Culture Collection (ATCC). All cells were maintained at 37 °C in a 5% CO 2 incubator and grown in 1640 medium (Gibco, Grand Island, NE, USA) supplemented with 10% FBS (Gibco, Grand Island, NE, USA). For BCR cross-linking and inhibition, 3 × 10 6 cells pre-treated with 4 μM R406 (Santa Cruz Biotechnology, Dallas, Texas, USA) or 1 uM ibrutinib (Selleck, Houston, TX, USA) for 60 min or equivalent DMSO vehicle, were stimulated by 10 μg/mL rabbit anti-human IgM (Jackson ImmunoResearch Laboratories, West Grove, PA USA) and goat anti-human IgG (Jackson ImmunoResearch Laboratories, West Grove, PA USA). MYC inhibitor 10058-F4 (Selleck, Houston, TX, USA) was used at 50uM.
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!