PMA (Sigma-Aldrich, P8139) was added to media at 25 ng ml−1 at 37 °C with 5% CO2 for denoted time. JQ1 (Tocris, 4499) was added to media at 0.5 μM at 37 °C with 5% CO2 for 4 h unless denoted otherwise. Triptolide (Tocris, 3253; Selleckchem, S3604) was added to media at 1 μM at 37 °C with 5% CO2 for 4 h unless denoted otherwise. Flavopiridol (Selleckchem, S1230) was added to media at 1 μM at 37 °C with 5% CO2 for 1 h unless denoted otherwise. Dox (MP Biomedicals, 198955) was added to media at 1 μg ml−1 at 37 °C with 5% CO2 for 24 h unless denoted otherwise. The splicing inhibitor, Plad B (ref. 54 (link); Tocris, 6070), was added to media at 5 μM at 37 °C with 5% CO2 for 2 h unless noted otherwise.
Plad b
Manufactured by Bio-Techne
Plad B is a laboratory equipment product from Bio-Techne. It is a device used for performing electrophoresis, a fundamental technique in molecular biology and biochemistry to separate and analyze macromolecules such as proteins or nucleic acids.
Automatically generated - may contain errors
Lab products found in correlation
Triptolide, by Selleck Chemicals (1 mentions)
Flavopiridol, by Selleck Chemicals (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Ifn α2, by BioLegend (1 mentions)
3p hprna, by InvivoGen (1 mentions)
Tlrl hprna, by InvivoGen (1 mentions)
Lyec 1, by InvivoGen (1 mentions)
Tnf α, by BioLegend (1 mentions)
2 protocols using plad b
1
Cell Treatment with Chemical Modulators
2
Cell-Based Cytokine Induction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plad B was purchased from Tocris (6070), and spliceostatin analog PF-06437837 (thailanstatin A methyl ester) was synthesized by Pfizer as previously reported (22 (link)). Cells were seeded in 96-well plates at 5000 cells/well (4T1), 10,000 cells/well (A549, CT26, and MC38 HEK), and 20,000 cells/well (B16F10) overnight in 75 μl media. The next day, 75 μl of a 2× solution of the indicated compound was added to wells, and cells were incubated at 37 °C until the indicated time points. Assays with THP1 cells were performed similarly except plating cells (100,000 cells) immediately before the addition of indicated compounds. Supernatant was removed and used in subsequent assays. 3p-hpRNA (Invivogen; tlrl-hprna) was prepared as a 2× solution in complex with the transfection reagent LyoVec (Invivogen; lyec-1) according to the manufacturer's instructions. After complexing, 75 μl of the 2× concentrated 3p-hpRNA/LyoVec was added to cells until the indicated time points. Positive control cytokines used were IFN-α2 (Biolegend; 592702) and TNFα (Biolegend; 570102).
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!