生物化学技术

问：我想做肌肉组织的乙酰胆碱酯酶染色，没有详细的步骤及注意事项，请熟悉的前辈们给予指点，不胜感激！ 答：1、动物麻醉后4％多聚甲醛（0.1M PB配制）固定。 2、取肌肉组织，用灌注液后固定3h。 3、若做冰冻切片，10％，20％，30％蔗糖脱水，用0.1M PB配制蔗糖。若做石蜡切片，梯度酒精脱水，二甲苯透明，石蜡包埋。 4、冰冻切片或石蜡切片。 5、石蜡切片经脱水后入0.01M PBS，冰冻切 ...

问：谁有a一醋酸萘酯酶（ANAE)染色法的具体步骤，我们这里查不到，还望各位大侠不吝赐教！ 答：中性非特异性脂酶(α—NAE)染色 酯酶染色的方法很多，根据与pH底物等不同作用，将常用的酯酶染色法分为两大类，即特异性酯酶染色和非特异性酯酶染色。后者又可分为3类：①中性非特异性酯酶染色法：α—乙酸菜酚酯酶、乙酸菜酚酯酶和乙酸菜酚AS-D酯酶。②酸性 ...

问：请教肝酯酶的生理功能及其测定方法？ 答：肝脂酶（hepaticlipase或hepatiltriglyceridase或hepaticendothelaillipase，HL或HTGL）属于与血液循环中内源性TG代谢有关的酶之一，与LPL在功能上有相似之处，然而却是两种不同性质的酶。 其特点是：①HL活性不需要ApoCⅡ作为激活剂；②SDS可抑制HL活性，而不受高盐浓度及鱼精蛋白的抑制；③主要 ...

问：磷酸二酯酶（phosphodiesterase，PDE）。请教：目前发现的PDE有多少个类型，共多少个亚型。其中阴茎海绵体组织中的PDE主要存在那几种类型及亚型。各种类型主要参与调节cGMP还是cAMP含量？（并给出出处－参考文献） 答：目前磷酸二酯酶可分类为11型，PDE1至PDE11。具体请参考下面文献： ...

问：本人想写一篇关于有机磷中毒的文章，可是胆碱酯酶的测定是用KU/ml来表达，而有机磷中毒的分级却用百分比来表示，其中有换算公式吗？ 答：中毒分级 a. 轻度中毒: 有头晕、头痛、恶心、呕吐、多汗、胸闷、视物模糊、无力等症状瞳孔可能缩小。全血胆碱酯酶活性一般为50%～70% 。 b. 中度中毒: 上述症状加重尚有肌束颤动、瞳孔缩小、轻度呼吸困难、流涎、腹痛、腹泻、步态蹒跚、意识清或模糊。全血胆碱酯 ...

问：抽提细胞总蛋白时，如何将脂类从蛋白中去除？如果目的蛋白可能是脂结合的膜蛋白。 答：作过脂阀研究的人都知道把脂类从细胞蛋白中分离出来非常困难。要用超级离心机，100000转以上离心几个甚至十几个小时，有点象提细胞膜蛋白的提取。还有一种分级提取的方法，也很烦琐。到目前似乎还没有简单的方法祛除脂类。 答：膜蛋白一般都有疏水性的基团，可以通过它的水溶性来去除脂质物质。为了提高疏水蛋白在水中的水溶性， ...

Srinivas Kodali Andrew Galgoci Sheo Singh Dr. Jun Wang Dr. jun_wang2@merck.com Merck Research Laboratories Related Journal & Article InformationManuscript: 2006-01-00990C Journal: Nature Article T ...

Lipid analysis During this week you will analyze the fatty acid composition of the individual lipid fractions recovered from the TLC plate. Gas chromatography is a very sensitive method for the separa ...

Lipid analysis Thin layer chromatography is based on the separation of a mixture of compounds as it migrates with the help of a suitable solvent through a thin layer of adsorbent material which has be ...

REFERENCES: Nikaido and Roseenberg (1983) J.Bact 153: 241-252; Yoshimura et al. (1983) J.Bact 258: 2308-2314. METHOD: 6.2 µmoles of lipid (ideally egg phophatidylcholine or dipalmitoyl phosphory ...

OBJECTIVE: Method for incorporating proteins into liposomes for liposome swelling assay or as an alternative antigen presentation method. REAGENTS: Dioleoyl Phosphatidyl Choline Buffer of choice / dis ...

EXTRACTION AND ANALYSIS OF LIPIDS The lipophorin molecule that we isolated from locust hemolymph functions as a re-usable lipid shuttle. HDLp has a density of approx. 1.08 g/ml and contains less lipid ...

Hancock Laboratory MethodsDepartment of Microbiology and ImmunologyUniversity of British Columbia British Columbia Canada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=41 OBJECTIVE: To methylate ...

Hancock Laboratory MethodsDepartment of Microbiology and ImmunologyUniversity of British Columbia British Columbia Canada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=59 REAGENTS: CHCl3:MeOH (2 ...

Contributor: Suprya JayadevDate: August 21 1991 NOTE - Use arachidonic acid from Biomol (# FA-003) 1) Make up the appropriate concentration of AA in 100% ethanol. 2) Add 1µl of phenol red soluti ...

Contributor: Suprya JayadevDate: Mar. 11 1991 Packing column: 1) To 20 g of 100-200 mesh Bio-Sil A silica gel add 80 mls of chloroform. 2) Place a small portion of glass wool at the base of the column ...

Contributor: Suprya JayadevDate: November 4 1991 1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum. 2) Pellet cells and wash 1 time with room temperature PBS. 3) Resuspend ...

Contributor: Suprya JayadevDate: Mar. 11 1991 I. Lower chloroform phase: 1) Dry on rotovapor system with house vacuum lines. It is not necessary to dry sample completely but sufficiently to yield a fi ...

Contributor: Suprya JayadevDate: Nov. 10 1993 1) Incubate cells with 1 µCi/ml of 3H-galactose for 72 hours.---> If treatment is for an extended period of time: treat in serum free media conta ...

Contributor: Suprya JayadevDate: Mar. 11 1993 Synthesis reaction: 1) Mix purified DMSM with cyclohexylamine and 14CH3I at a ratio of 1/1.1/1.3 in 5 ml of methanol. 2) Allow reaction to proceed at room ...