多聚赖氨酸的配置、保存、使用.doc

多聚赖氨酸的配置、保存、使用 一、常用的多聚赖氨酸包被可以用三蒸水，或者PBS，浓度一般为用0.1?mg/ml进行包被。 二、其他常用包被方法比较(以各种免疫分析为例)snap 三、我配成1mg/ml的储存液，用Mini-Q(超纯水)配制，放在－20储存，使用时稀释10倍（也用超纯水），即终浓度100ug/ml，过滤后使用。工作液过滤使用，如一次用不完，放在4储存。 多聚赖氨酸在水溶液中易分解，所以一次不要配太多工作液。 四、我现在要配多聚赖氨酸,书上写的用?PBS配,但没写PH,浓度，向各位请教。 ??????答：PH应该在7.0~7.4，PBS：取氯化钠7.650?g，无水磷酸?氢二钠0.724?g，磷酸二氢钾0.210g?溶于蒸馏水1000?mL?中，以1N?氢氧化钠?溶液调pH?值为7.0~7.4，经121灭菌15?分钟 五、我准备用多聚赖氨酸涂片培养细胞,不知道多聚赖氨酸涂过的玻片如何灭菌？能否干烤灭菌？ ???????答：Sigma的产品说明书上写的很明白的。 ???????另外，有本书上是这么写的，供参考： Poly-lysine-coated?tissue?culture?surfaces ????????To?coat?coverslips:?Prepare?a?stock?solution?by?dissolving?25?mg/ml?polylysine?in?4.73?ml?water?(both?poly-L-lysine?and?poly-D-lysine?are?used?to?coat?tissue?culture?surfaces;?check?specific?protocol?for?choice?of?isomer)?and?filter?sterilize?through?a?0.22-μm?filter.?Store?in?100-μl?aliquots?at??20°C.?When?ready?to?use,?dilute?one?aliquot?in?40?ml?water?to?prepare?13?μg/ml?working?solution.?Sterilize?coverslips?by?autoclaving?prior?to?coating.?Dip?coverslips?in?the?working?solution,?then?incubate?15?min?to?several?hours?in?a?humidified?37°C,?5%?CO2?incubator.?Allow?surface?to?dry. ???????To?coat?culture?dishes?or?8-well?chamber?slides:?Prepare?a?stock?solution?by?dissolving?100?mg?poly-lysine?in?100?ml?water?(both?poly-L-lysine?and?poly-D-lysine?are?used?to?coat?tissue?culture?surfaces;?check?specific?protocol?for?choice?of?isomer)?and?filter?sterilize?through?a?0.22-μm?filter.?Store?in?5-ml?aliquots?at??20°C.?When?ready?to?use,?dilute?1?part?stock?solution?with?9?parts?water?to?prepare?100?μg/ml?working?solution.?Fill?tissue?culture?dishes?or?slide?wells?with?the?working?solution?and?incubate?1?hr?in?a?humidified?37°C,?5%?CO2?incubator,?then?remove?solution?by?vacuum?aspiration?and?allow?surface?to?dry. Store?coated?tissue?culture?ware?up?to?3?months?at?4°C.?Use?diluted?solutions?only?once,?but?unused?diluted?aliquots?can?be?stored?up?to?3?months?at?4°C. ???????你可以配置好PLL后单独将其过滤除菌，分装，待用。玻片也是事先泡洗洁精，泡酸，双蒸水洗，烘干后高压灭菌，待用。培养的前一天包被培养板时先把玻片放