UN 3316 9
188.6 °F - closed cup
87 °C - closed cup
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
The use of detergent will lead to maximum protein yield. However we find that the hypotonic lysis is extremely effective and in addition for very fragile cells we offer an isotonic lysis option.
Several references describe the use of the N-XTRACT kit in sample preparation for EMSA work.1. Xie, J., et al., β2-Microglobulin as a negative regulator of the immune system: high concentrations of the protein inhibit in vitro generation of functional dendritic cells. Blood, 101(10), 4005-4012 (2003).2. Marambaud, P., et al., A CBP Binding Transcriptional Repressor Produced by the PS1/ε-Cleavage of N-Cadherin Is Inhibitedby PS1 FAD Mutations. Cell, 114, 635-645 (2003).3. Chen, G., et al., Phosphorylated FADD induces NF-κB, perturbs cell cycle, and is associated with poor outcome in lung adenocarcinomas. Proc. Nat. Acad. Sci. USA, 102(35), 12507-12512 (2005).
We have used the kit with several tissues: Mouse lung and brain, Rat liver and Rabbit muscle. For these tissues we usually used the kit with 100 mg tissue. After extraction with 140 μl Extraction buffer, we obtained an average yield of protein concentration as follows: 3.2 mg/ml for brain, 4.1 mg/ml for lung, 3.3 mg/ml for muscle and 10 mg/ml for liver.
We recommend the use of a glass tissue homogenizer, with a type B pestle, or alternatively a syringe with a narrow-gauge (No. 27) hypodermic needle may be used.
It is important to keep in mind that this kit will provide a crude nuclear fraction. Depending on the care taken at the step where the nuclei are pelleted and the supernatant containing the cytoplasmic fraction is removed, we have found that the cytoplasmic contamination of the nuclear pellet may be a few percent.
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
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