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泳道1含分子量标准(分子量标准物VII,BoehringerMannheim);泳道2含有引物T7:476-30(SEQIDNO:11)和引物485-26:(SEQIDNO:12)扩增约2.1kb的PCR片段之后得到的PCR产物;泳道3含分子量标准(分子量标记物II,BoehringerMannheim)。图6显示了包含部分GlcNAcT-V编码顺序的几种序列,此处命名为扩增物顺序(amplimersequence),...
After 15 min, the mixtures were incubated with 5 μl of 100 mM iodoacetamide at room temperature for 15 min (34). The mixtures were diluted with 140 μl of water (34), and then each was incubated with 9 μl of 0.2 mg/ml of sequencing grade trypsin (Boehringer Mannheim) at 37°C ...
with or without 25 μM aurine-tricarboxylic acid (ATA), the endonuclease inhibitor (Sigma, St. Louis, MO, USA), were collected, from the same wells used for MTS determination, and LDH activity evaluated by a LDH cytotoxicity detection kit (Boehringer-Mannheim, Mannheim, Germany), according to...
N-glycans were released from glycoproteins in the serum samples by digestion with N-glycosidase F (PNGase F; Roche Diagnostics GmbH, Mannheim, Germany) in-gel blocks as described previously [15]. Briefly, serum samples were reduced and alkylated and then set in sodium dodecyl sulfate gel blocks...
Inflammation in the air pouch was induced by local injection of recombinant murine TNF-α (20 ng) (Boehringer Mannheim, Indianapolis, Ind.) dissolved in 100 μl of sterile PBS. While the mice were anesthetized with isoflurane, the air pouches were lavaged twice with 3 ml of sterile PBS 4...
PCR reactions were done using the Taq polymerase system (Boehringer Mannheim, Mannheim, Germany). PCR reactions were performed in a total volume of 50 μl, containing 1× Taq PCR reaction buffer, 0.25 mM DNTP, 0.5 μM of primers MOUSE-GFRα4-sp2 and MOUSE-GFRα4-ap2, 1 μl of Taq...