The Bacterial Cell Lysis Kit has been developed for the extraction of biologically active, soluble proteins and inclusion bodies from bacterial cells. The Bacterial Lysis Buffer is a proprietary improvement on the lysozyme based lysis method, which allows extraction of soluble proteins and concurrent removal of nucleic acids (DNA & RNA) released during cell lysis. The kit’s lysis eliminates viscosity build-up, allowing effective clarification with lower centrifugal forces.

Bacterial Cell Lysis Buffer is based on organic buffering agents andutilizes a mild non-ionic detergent and a proprietary combination ofvarious salts and agents to enhance extraction and stability ofproteins. Depending on the application, additional agents such asreducing agents chelating agent, and protease inhibitors may be addedinto Bacterial Cell Lysis Buffer. This reagent has been tested for usewith several widely used bacteria including E. coli strains. Bacterial Cell Lysis Buffer eliminates the need for laborious mechanical lysis of bacterial cells and removal of DNA/RNA with nuclease treatments. The proprietary combination of this reagent provides a simple and versatile method of bacterial protein extraction and isolation of inclusion bodies.

The Bacterial Cell Lysis Kit (GB-176; 100, 250 or 500 preps) includes a separate tube of modified lysozyme preparation that contains nucleases and results in optimal lysis and minimal contamination. This kit contains all of the reagents needed to perform protein extraction.For extracting soluble proteins, one prep is suitable for extracting approximately 300 mg wet cell pellets for every 1 ml Bacterial Cell Lysis Buffer. When the kits are used for extracting soluble proteins from spheroplasts, one prep is suitable for extracting approximately 90 mg wet cell pellets for every 40 μl Bacterial Suspension Buffer. Bacterial Lysis Buffer is also available separately for further downstream applications.

Storage/Handling: Store at 4°C. Do NOT freeze. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.

小鼠 1.       首先确定针的入口点。 2.       在膝关节的上部横跨腹部画一条想像的线。 3.       针应该沿着动物右侧这条线并贴近中间插入。（对于雌鼠，针的入口点位于最后一个乳头稍靠中间的部位。） 4.       针从小鼠的右侧插入避开盲肠，盲肠是腹部左侧一个大的充满液体的器官。小肠（位于右侧）不大可能被针刺穿。 5.       针的插入从插入点离尾部或横向太远如上显示将是危险的，可能会注射到后腿里，这样将会造成肌肉组织受伤。 6.       为完成腹腔注射，必须很好地控制好小鼠以防在操作过程中移动。一旦针插入到腹部避免器官受损伤。 7.       控制好小鼠并倾斜以便使其头向下，腹部露出来。 8.       用70%乙醇对注射位点进行彻底消毒。 9.       对于腹部注射我们建议用25-27 gauge直径的针，以30度角度插入腹部。 10.   针应插入大约半厘米（4-5mm）的深度。 11.   轻轻地吸出以确认针没有穿透血管，肠或膀胱。    a.       褐绿色吸出物表示针插进了小肠。    b.       黄色吸出物暗示针插进了膀胱。 12.   如果任何液体被吸出，表示您的溶液被污染了必须弃掉并用一个新的注射器和针头重新操作。 13.   如果没有液体被吸出，你可以开始注射。 14.   取出针头并记录下注射的时间，以确定信号达峰时间。 大鼠 按照小鼠腹腔注射操作流程。对于大鼠我们建议用一个25 gauge 直径的针头注射。 注意 l  确保针是倾斜的。 l  使动物的头倾斜向下为了使肠内物向下移动，以保持腹部四分区下部为空腔。 l  为了精确注射深度防止深层注射，我们建议放一根导管在针头上留出4-5mm为合适的深度。 l  确保注射体积，我们建议小鼠不超过2ml，大鼠不超过5ml（请参看附录关于不同动物和位点的不同注射体积。） l  给大鼠注射时，用布盖在大鼠的头部防止小鼠和研究人员紧张。 静脉注射