生物化学技术

CD44变异体V6、V7/8在非小细胞肺癌中的表达及临床意义　　第三军医大学学报2000年第22卷第7期孙焰　陈幸华　罗平　贺光友　　摘　要：目的　探讨CD44 v6、V7/8与非小细胞肺癌(Non-small cell lung cancer,NSCLC)患者的淋巴结转移及预后的关系。方法　采用免疫组化方法观察CD44 v6、V7/8在76例原发性非小细胞肺癌组织及转移淋巴结中的表达。结果　在肺癌中CD44 v6、V7/8表达鳞癌多于腺癌(P＜0.01)；在正常肺组织中无表达。 ...

免疫组织化学的概念： 免疫组化是利用抗原与抗体特异性结合的原理，通过化学反应使标记抗体的显色剂 (荧光素、酶、金属离子、同位素) 显色来确定组织细胞内抗原(多肽和蛋白质)，对其进行定位、定性及定量的研究，称为免疫组织化学。免疫组化实验中常用的抗体为单克隆抗体和多克隆抗体。免疫组化实验所用的组织和细胞标本有哪些？实验所用主要为组织标本和细胞标本两大类，前者包括石蜡切片（病理大片和组织芯片）和冰冻切片，后者包括组织印片、细胞爬片和 ...

免疫组化-石蜡切片的染色1、载玻片的处理：抗原修复过程中，由于高温、高压、辐射等诸多因素的影响，极易造成脱片。为保证试验的正常进行，可选用我公司提供的ZLI-9001 APES、ZLI- 9003 HistogripTM或ZLI-9005 Poly-L-Lysine等几种试剂，对已清洗的载玻片进行处理。具体方法如下：1.1 APES：现用现配。将洗净的玻片放入以1:50比例丙酮稀释的APES中，停留20~30秒钟，取出稍停片刻，再入纯丙酮溶液 ...

免疫组化的经验总结--操作要点和技巧1 固定：最好用4%的多聚甲醛固定液。对于冰冻切片，甲醛固定有时比冰冻丙酮好；但对于不同的组织和抗原，可选用不同的固定液。 有时候商品化的抗体会有比较适合而推荐的固定液，请于购置前注意说明书。 Bouin S固定液：饱和苦味酸750ml，甲醛250ml，冰醋酸50ml，其对组织的穿透力较强，固定较好，结构完整，但因偏酸，对抗原有一定损害，且组织收缩明显，不适于组织标本的长期保存。 PLP液：即高碘酸钠-赖 ...

Your antigen should at least have a molecular weight of 20 KDa for development of a strong immune response in animals. If it is a small protein, it can be linked to a larger protein by chemical methods or cloned and expressed as a fusion protein with a larger protein. If it is a peptide or small molecule, a carrier-hap ...

make an Antibody against a Peptides If your peptide is synthesized with a terminal cysteine residue (i.e., a free sulfhydryl group, -SH), you can use any maleimide-activated carrier protein to create an immunogen. One-step chemical reactions are easy with Pierce Sulfo-SMCC Pre-Activated C ...

The production of monoclonal antibody by hybridoma fusion: Immortalization of sensitized B lymphocytes from immune mice.

Immunization Protocol for Monoclonal Antibody ProductionAntigens:Proteins: 10-100 µgCells: 0.5 - 5.0 x107Adjuvant:Complete Freunds AdjuvantNoneSchedule:Day Manipulation Adjuvant Site 0Primary ImmunizationCFAs.c.14Boost #1IFAs.c.24Boost #2IFAs.c.35 ...

天然疫苗和人工疫苗 本综述概述了细胞和抗体介导的免疫保护及免疫记忆的基本机制,以此为基础,用抗原在免疫保护中的作用来评估疫苗研究的成功和困难,人类在出生后12~48个月期间,来自母乳和血清的获得性抗体可避免儿童期常见病和其它感染性疾病.基于此事实,形成了这样一种概念:获得性抗体骤减了婴幼儿的各种感染,所以使它们变成了有效的疫苗.如果没有这种被动保护条件下的"天然疫苗",除非婴幼儿在早期有效接种了各种疫苗且免疫系统 ...

Choline Labeling and TNF Treatment of HL-60 CellsContributor: Suprya JayadevDate: Aug. 6, 19911) 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 final pellet in an appropriate volume of prewarmed serum f ...

一)制备工艺流程（1）脊髓灰质炎病毒活疫苗制备工艺猴 肾 细 胞 培 养←接种病毒（测0%正常细胞对照）收获病毒，合并，加MgCl2→无菌试验→←猴病毒检查（获肾细胞）（SV40）B病毒检查（兔肾细胞）→←病毒效价滴定←合并，过滤加MgCl2←无菌试验分亚批→←柯萨奇病毒检查（乳鼠）病毒滴定→←型特异性检查B病毒复检（家兔）→←T特征试验分装病毒滴定→←猴体残余致麻痹力试验（安全试验）病理切片结核杆菌检查→分装→←无菌试验病毒滴定→→保存于-20℃待检定合格后加 工 糖 ...

Sphingomyelin Quantitation Post-choline Labeling of HL60 CellsContributor: Suprya JayadevDate: Jun. 16, 1992ReagentsTMT buffer:0.19 M Tris-HCl (pH 7.4-7.6)12 mM MgCl20.2 % Triton X-100SMase solution:SMase from Streptomyces sp. @ 2 units/ml in 10 mM Tris-HCl (pH 7.4-7.6)Lipid Extra ...

Gel-elongation assay for type II fatty acid synthesisSrinivas Kodali Andrew Galgoci Sheo Singh Dr. Jun Wang Dr., jun_wang2@merck.com, Merck Research Laboratories Related Journal & Article InformationManuscript: 2006-01-00990CJournal: NatureArticle Title: Platensim ...

AA & Metabolite Quantitation of Cell Pellets Post-AA Labeling Contributor: Suprya JayadevDate: November 4, 1991Extraction:1) Following spin, save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediately upon removing ...

AA & Metabolite Quantitation of Media Post-AA Labeling Contributor: Suprya JayadevDate: November 4, 19941) Remove 2 500 µl aliquots of supernatant into scintillation vials, add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant into each of two pyrex glass tubes ...

Analysis of Oligosaccharide Ligands by High Performance Liquid Affinity ChromatographyWeiTong Wang~GlycoTech Corporation, Rockville, Maryland 20850 High performance liquid affinity chromatography (HPLAC) is a useful technique for investigation of the interacti ...

Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC PlatesRonald L. Schnaar~Professor, Johns Hopkins University Medical School, Baltimore, Maryland 21205This procedure allows detection of specific cell adhesion to glycolipids resolved on TLC p ...

A Method for Micro-Scale Isolation and Purification of GangliosidesStephan Ladisch~Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C. 22010This solvent partition method permits the isolation of gangliosides from sm ...

Many different molecules have been described to promote cell adhesion including several cell surface carbohydrate-binding proteins. Measuring cell adhesion in the convenient 96-well microtiter plate format is difficult due to the shear forces generated by washing the wells. This ...

Dynamic Flow Assay in a Parallel Plate Flow ChamberJohn T. Patton~GlycoTech Corporation, Rockville, Maryland 20850Flow assays allow visualization of cell adhesion under well-defined wall shear stress. The visualization of the different events of cell adhesion can be quantified by ...