Native ChIP was performed with 1 × 107of cells. Cells were resuspended in cold douncing buffer (10 mM Tris-Cl pH7.5, 4 mM MgCl2, 1 mM CaCl2, 1X PIC) and homogenized 20 times with 25 gauge 5/8 inch needle. 0.8 uL of MNase (Thermo Scientific™, 88216) was added to ...
Samples were incubated for 2 h at 4 °C, and subsequently washed with the binding buffer at least five times. The pulled‐down proteins by Glutathione Sepharose 4B beads were detected by immunoblotting with anti-MBP antibody (Sigma, M6295). Co-immunoprecipitation Total protein from the ...
classLineBuffer(LineSingle):'''LineBuffer defines an interface to an "array.array" (or list) in whichindex 0 points to the item which is active for input and output.Positive indices fetch values from the past (left hand side)Negative indices fetch values from the future (if the array...
After binding to concanavalin A–coated magnetic beads (ConA beads), bead-bound cells were incubated with anti-V5 antibody (Abcam, ab15828, 5ug/1:10) or normal rabbit IgG (CST, 2729, 0.5ug/1:100) for 2 h at room temperature. After brief wash with dig-wash buffer, cells were then...
Samples were then washed four times with blocking buffer, followed by incubation with secondary antibodies in blocking buffer at room temperature for 1 h. Slides were then mounted with Prolong Diamond Antifade Mountant (Thermo Fisher) and analyzed after 24 h of curing. Of note, for visualization...
The matrix beads were washed three times with immunoprecipitation buffer and twice with 20 mM Tris-HCl, pH 7.5, 150 mM NaCl, and 5% glycerol. All of the wash steps were performed at 4°C for 10 min on a rotary shaker followed by centrifugation at 1500 rpm. The immunoprecipitated proteins...
The binding of Rat2 cells nuclear proteins to the P1 minimal promoter region was confirmed by EMSA analysis. We also showed the existence of a minimal promoter region within the sequence -720/-537 upstream the ATG for transcript T2 and a strong positive regulation region in the exon E-3. ...
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Notch signaling is highly conserved in the metazoa and is critical for many cell fate decisions. Notch activation occurs following ligand binding to Notch extracellular domain. In vitro binding assays have identified epidermal growth factor (EGF) repeats
The core computational unit, which includes control, arithmetic, registers and typically some cache, is replicated some number of times and connected to memory via a network. As a result, all modern processors require parallel code in order to achieve good utilization of their computational power....