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人小细胞肺癌细胞 NCI-H446齐一生物科技(上海)有限公司是一家专业生产和销售各种生化试剂,细胞株、原代细胞、菌种、医药中间体,标准品和对照品的大型化学科技公司.齐一生物科技(上海)有限公司:.Web:www.qiyibio。。com
人小细胞肺癌细胞 NCI-H446细胞培养方法
1、收到细胞,请查看瓶子是否有破裂,培养基是否漏出,是否浑浊,如有请尽快。
2、收到细胞,如包装完好,请在显微镜下观察细胞。,由于运输过程中的问题,细胞培养瓶中的贴壁细胞有可能从瓶壁中脱落下来,显微镜下观察会出现细胞悬浮的情况,出现此状态时,请不要打开细胞培养瓶,应立即将培养瓶置于细胞培养箱里静止3-5小时左右,让细胞先稳定下,再于显微镜下观察,此时多数细胞会重新贴附于瓶壁。如细胞仍不能贴壁,请用台盼蓝染色法鉴定细胞活力,如台盼蓝染色证实细胞活力正常请按悬浮细胞的方法处理
3. 收到细胞时如无异常情况 ,请在显微镜下观察细胞密度,如为贴壁细胞,未超过80%汇合度时,将培养瓶中培养基吸出,留下 5-10ML培养基继续培养:超过80%汇合度时,请按细胞培养条件传代培养。如为悬浮细胞,吸出培养液,1000转/分钟离心3分钟,吸出上清,管底细胞用新鲜培养基悬浮细胞后移回培养瓶。
4,将培养瓶置于37℃培养箱中培养,盖子微微拧松。吸出的培养基可以保存在灭菌过的瓶子里,存放于4℃冰箱,以备不时之需。
5 、24小时后,细胞形态已恢复并贴满瓶壁,就可以传代了。(贴壁细胞)将培养瓶里的培养基倒去,加3-5ml(以能覆盖细胞生长面为准)PBS或Hanks’液洗涤后弃去。加0.5-1ml 0.25%含EDTA的胰酶消化,消化时间以具体细胞为准,一般1-3分钟,不超过5分钟。可以放入37℃培养箱消化。轻轻晃动瓶壁,见细胞脱落下来,加入3-5ml培养基终止消化。用移液管轻轻吹打瓶壁上的细胞,使之*脱落,然后将溶液吸入离心管内离心,1000rpm/5min。弃上清,视细胞数量决定分瓶数,一般一传二,如细胞量多可一传三,有些细胞不易传得过稀,有些生长较快的细胞则可以多传几瓶,以具体细胞和经验为准。(悬浮细胞)用移液管轻轻吹打瓶壁,直接将溶液吸入离心管离心即可。
6,贴壁细胞 ,悬浮细胞。严格无菌操作。换液时,换新的细胞培养瓶和换新鲜的培养液,37度 5%CO2 培养
7. 收到细胞后,请镜下观察细胞,用恰当方式处理细胞。若悬浮的细胞较多,请离心收集细胞,接种到一个新的培养瓶中。弃掉原液,使用新鲜配制的培养基,使用进口胎牛血清. 刚接到细胞,若细胞不多时 血清浓度可以加到15%去培养。若细胞迏到80%左右 ,血清浓度还是在10%。
培养基参考
500ml基础培养基(含:必需与非必需氨基酸、维他命、有机与无机成分、激素、生长因子与微量元素)、细胞生长添加剂、抗生素/抗菌素溶液或青霉素/链霉素(P/S)溶液、FBS,适合在饱和湿度为5%CO2与95%空气的培养箱中使用
Rat Stromal cell derived factor 1β,SDF-1β ELISA Kit
Rat angiopoietin receptor Tie1,ANG-R-Tie1 ELISA kit
Rat tyrosine kinase with immunoglobulin-like and EGF-like domains 2,Tie-2 ELISA kit
Rat tissue inhibitors of metalloproteinase 2,TIMP-2 ELISA Kit
Rat tissue inhibitors of metalloproteinase 3,TIMP-3 ELISA Kit
Rat tissue inhibitors of metalloproteinase 4,TIMP-4 ELISA Kit
Rat Tumor necrosis factor soluble receptor Ⅰ,TNFsR-Ⅰ ELISA KIT
Rat Tumor necrosis factor soluble receptor Ⅱ,TNFsR-Ⅱ ELISA KIT
Rat Tumor necrosis factor β,TNF-β ELISA KIT
Mouse L-Selectin ELISA kit
Rat tumor necrosis factor-related apoptosis-inducing ligand 1,TRAIL R1 ELISA Kit
Rat tumor necrosis factor-related apoptosis-inducing ligand 3,TRAIL R3 ELISA Kit
Rat tumor necrosis factor-related apoptosis-inducing ligand 4,TRAIL R4 ELISA Kit
Rat soluble tumor necrosis factor-related apoptosis inducing ligand,sTRAIL ELISA KIT
Rat urokinase plasminogen activator,uPA ELISA kit
Rat Urokinase-type Plasminogen Activator Receptor,PLAUR/uPAR ELISA kit
Rat Interleukin 12,IL-12/P70 ELISA KIT
Rat Vascular Endothelial cell Growth Factor B,VEGF-B ELISA kit
Rat Interleukin 12,IL-12/P40 ELISA KIT
Rat Vascular Endothelial cell Growth Factor C,VEGF-C ELISA kit
Rat Interleukin 11,IL-11 ELISA KIT
Rat Insulin-like growth factor binding protein 4,IGFBP-4 ELISA kit
Rat Insulin-like growth factor binding protein 3,IGFBP-3 ELISA kit
Rat Vascular Endothelial cell Growth Factor D,VEGF-D ELISA kit
Rat Insulin-like growth factor binding protein 2,IGFBP-2 ELISA kit
Rat Vascuoar endothelial cell growth factor receptor 1,VEGFR-1/Flt1 ELISA Kit
Rat Insulin-like growth factor binding protein 1,IGFBP-1 ELISA Kit
Rat Vascuoar endothelial cell growth factor receptor 2,VEGFR-2/Flk-1 ELISA kit
Rat hepatocyte growth factor,HGF ELISA kit
Rat glucoprotein 130,gp130 ELISA KIT
Rat growth hormone,GH ELISA KIT
Rat Granulocyte Colony Stimulating Factor,G-CSF ELISA Kit
Rat neutrophil activating protein-2,NAP-2 ELISA Kit
Rat FMS-like tyrosine kinase 3,Flt3 ELISA Kit
Rat FMS-like tyrosine kinase 3 ligand,Flt-3L ELISA Kit
Rat basic fibroblast growth factor 9,bFGF-9 ELISA Kit
Rat basic fibroblast growth factor 4,bFGF-4 ELISA Kit
Rat basic fibroblast growth factor 6,bFGF-6 ELISA Kit
Rat acidic fibroblast growth factor 1,aFGF-1 ELISA Kit
Rat Factor-related Apoptosis ligand,FASL ELISA Kit
Rat Factor-related Apoptosis,FAS ELISA Kit
Rat Erythropoietin receptor,EPOR ELISA Kit
Rat Eotaxin 1 ELISA Kit
Rat eosinophil chemotactic factor,ECF ELISA Kit
Rat Endocrine gland vascular endothelial growth factor,EG-VEGF ELISA Kit
Rat 1,3-βD glucosidase,1,3-βD-Glu ELISA Kit
Rat soluble endothelial protein C receptor,sEPCR ELISA Kit
Rat Cluster of differentiation 30,CD30 ELISA Kit
Rat Epithelial-Cadherin,E-Cad ELISA Kit
Rat beta cellulin,BTC ELISA Kit
Rat Angiopoietin 1,ANG-1 ELISA Kit
Rat Angiogenin,ANG ELISA Kit
Rat keratinocyte autocrine factor/Amphiregulin,KAF/AR ELISA Kit
Rat Activated Leukocyte Cell Adhesion Molecule,ALCAM ELISA Kit
Rat Interleukin 9,IL-9 ELISA KIT
Rat Macrophage Migration Inhibitory Factor,MIF ELISA Kit
Rat transforming growth factor α,TGF-α ELISA Kit
Rat Progesterone,PROG ELISA kit
Rat estrogen,E ELISA Kit
Rat Vascuolar cell adhesion molecule 1,VCAM-1 ELISA kit
Rat Interleukin 8,IL-8 ELISA Kit
Rat adiponectin,ADP ELISA Kit
Rat free chorionic gonadotropin,f-βCG ELISA KIT
rat aldosterone,ALD ELISA Kit
rat Noradrenaline,NA ELISA Kit
rat Prostaglandin F,PG-F ELISA Kit
rat Prostaglandin E1,PG-E1 Elisa kit
rat Endothelin 1,ET-1 ELISA Kit
Rat bradykinin,BK ELISA Kit
rat Resistin ELISA Kit
rat prolactin,PRL ELISA Kit
rat growth hormone releasing hormone,GHRH ELISA Kit
rat follicle-stimulating hormone,FSH ELISA Kit
rat Estradiol receptor,ER ELISA Kit
Rat Pro Atrial Natriuretic Peptide,Pro-ANP ELISA Kit
Rat Big Endothelin,Big ET ELISA Kit
Rat Big Endothelin,Big ET ELISA Kit
Rat very late appearing antigen,VLA ELISA Kit
Rat Calcitonin,CT ELISA Kit
磷酸酶抑制剂复合物 III 10X
Bacterial Activity-Keeping Lysis Buffer细菌活性化蛋白制备裂解液
Menbrane Protein Dissolution Buffer膜蛋白溶解液
真菌和酵母用蛋白酶抑制剂复合物 100X
细菌用蛋白酶抑制剂复合物 100X
Tissue Total Protein Lysis Buffer组织蛋白裂解液
5x蛋白印迹膜再生液(强)
Inclusion body Purgation buffer包涵体洗涤液
Western and IP Cell lysis BufferWestern及IP细胞裂解液
5x蛋白印迹膜再生液(温和)
2D Sample Extraction Buffers I(major components: urea,NP-40 )2D电泳样品提取液 I
EZ-Buffers 10xHigh Molecules Western Transfer Buffer (no methanol)缓冲液
无蛋白封闭液 2% 封闭试剂
2D Sample Extraction Buffers II(major components: urea,CHAPS )2D电泳样品提取液 II
2D Sample Extraction Buffers V(major components: urea,thiourea, CHAPS )2D电泳样品提取液 V
2D Sample Extraction Buffers III(major components: urea,thiourea, CHAPS, ASB-16 )2D电泳样品提取液 III
2D Sample Extraction Buffers IV(major components :urea,thiourea ,CHAPS, SB3-10 )2D电泳样品提取液 IV
2D Sample Extraction Buffers VI(major components: urea,thiourea, CHAPS, ND SB201 )2D电泳样品提取液 VI
EZ-Buffers B 10XTBS Buffer缓冲液
EZ-Buffers C 10Xwestern transfer buffer缓冲液
无蛋白封闭液 1%封闭试剂in 1XTBS
EZ-Buffers E 10XPBST buffer with Tween缓冲液
EZ-Buffers D 10XPBS buffer solution缓冲液
EZ-Buffers I 2x Primary/secondary antibody dilution buffer soluition (in PBS)缓冲液
EZ-Buffers J 2xWestern block buffer in PBS with non-fat milk缓冲液
EL-P-NPP Chromogenic Reagent kit
EZ-Buffers K 5xAP( Alkaline phosphatase) buffer solution 缓冲液
EL-OPD Chromogenic Reagent kit
EL-TMB显色试剂
EL- OT Chromogenic Reagent kit
EL- ABTS Chromogenic Reagent kit
EZ-Buffers H 10XTBST buffer solution缓冲液
Protein Stains C: Quick& Sensitive Reversible Stain
Protein Stains A: Rapid Coomassie Stain Solution快速考马斯亮蓝染色试剂
Protein Stains B: Ultra-Sensitive Stain超敏染色液
SB3-10
SB3-10
SB3-12
SB3-12
SB3-14
SB3-14
SB3-16
SB3-16
SB3-18
SB3-18
Sulfo DST
Sulfo EGS
Sulfo-MBS
SMCC
Sulfo-SMCC
SPDP
Sulfo EMCS
EZ-Buffers Q 1xTris-Caps transfer buffer solution (Anode buffer 500ml + Cathode buffer 500ml)缓冲液
SefinoseTM 4B 琼脂糖凝胶
SefinoseTM 6B 琼脂糖凝胶
SefinoseTM CL-4B 琼脂糖凝胶
SefinoseTM CL-4B 琼脂糖凝胶
SefinoseTM CL-6B琼脂糖凝胶
SefinoseTM CL-6B琼脂糖凝胶
SefinoseTM 4 Fast Flow 琼脂糖凝胶
SefinoseTM 4 Fast Flow 琼脂糖凝胶
SefinoseTM 6 Fast Flow琼脂糖凝胶
SefinoseTM 6 Fast Flow琼脂糖凝胶
Q SefinoseTM FF琼脂糖凝胶
Q SefinoseTM FF琼脂糖凝胶
DEAE SefinoseTM琼脂糖凝胶
DEAE SefinoseTM琼脂糖凝胶
SP SefinoseTM FF琼脂糖凝胶
SP SefinoseTM FF琼脂糖凝胶
CM SefinoseTM琼脂糖凝胶
CM SefinoseTM琼脂糖凝胶
Q SefinoseTM琼脂糖凝胶
Q SefinoseTM琼脂糖凝胶
SP SefinoseTM琼脂糖凝胶
SP SefinoseTM琼脂糖凝胶
Q SefinoseTM HP琼脂糖凝胶
Q SefinoseTM HP琼脂糖凝胶
SP SefinoseTM HP琼脂糖凝胶
SP SefinoseTM HP琼脂糖凝胶
Butyl SefinoseTM 4 Fast Flow丁基疏水色谱柱料
Butyl SefinoseTM 4 Fast Flow丁基疏水色谱柱料
Phenyl Sefinose 6 Fast Flow (low sub,LS) 标准载量苯基疏水色谱柱料