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文献和实验Mouse B cell isolation with magnetic beads
and incubate on ice for 15min; l Negative : 1.Resuspend cells with MACS to 400 ul and 80 ul B cell Biotin each spleen,incubate on ice for 15 min; 2. Add 300 MACS and 200 anti-biotin Beads each spleen, incubate on ice for 15 min.
Dual- and Triple-Color Fluorospot
in the analysis, e.g., IL-10, rat mAb 9D7 is used for coating and tag (different from FITC and biotin)-labeled rat mAb 12G8 is used for detection. All mAbs from Mabtech, Nacka Strand, Sweden. 4. Secondary reagents for amplification: Mouse anti-FITC mAb
or HP1s associated with 5' biotin labeled siRNA EF52. Schematic methodology is shown for detection from cell lysates of biotin labeled siRNAs in complex with flag-tagged DNMTs or HP1s (alpha, beta, or gamma). B) Triple immunoprecipitation assay
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