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當前位置:齊一生物科技(上海)有限公司>>細胞生物>>原代細胞>> 人小細胞肺癌細胞 NCI-H446
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更新時間:2015-11-20 16:01:56瀏覽次數(shù):338次
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人小細胞肺癌細胞 NCI-H446細胞培養(yǎng)方法
1、收到細胞,請查看瓶子是否有破裂,培養(yǎng)基是否漏出,是否渾濁,如有請盡快。
2、收到細胞,如包裝完好,請在顯微鏡下觀察細胞。,由于運輸過程中的問題,細胞培養(yǎng)瓶中的貼壁細胞有可能從瓶壁中脫落下來,顯微鏡下觀察會出現(xiàn)細胞懸浮的情況,出現(xiàn)此狀態(tài)時,請不要打開細胞培養(yǎng)瓶,應立即將培養(yǎng)瓶置于細胞培養(yǎng)箱里靜止3-5小時左右,讓細胞先穩(wěn)定下,再于顯微鏡下觀察,此時多數(shù)細胞會重新貼附于瓶壁。如細胞仍不能貼壁,請用臺盼藍染色法鑒定細胞活力,如臺盼藍染色證實細胞活力正常請按懸浮細胞的方法處理
3. 收到細胞時如無異常情況 ,請在顯微鏡下觀察細胞密度,如為貼壁細胞,未超過80%匯合度時,將培養(yǎng)瓶中培養(yǎng)基吸出,留下 5-10ML培養(yǎng)基繼續(xù)培養(yǎng):超過80%匯合度時,請按細胞培養(yǎng)條件傳代培養(yǎng)。如為懸浮細胞,吸出培養(yǎng)液,1000轉(zhuǎn)/分鐘離心3分鐘,吸出上清,管底細胞用新鮮培養(yǎng)基懸浮細胞后移回培養(yǎng)瓶。
4,將培養(yǎng)瓶置于37℃培養(yǎng)箱中培養(yǎng),蓋子微微擰松。吸出的培養(yǎng)基可以保存在滅菌過的瓶子里,存放于4℃冰箱,以備不時之需。
5 、24小時后,細胞形態(tài)已恢復并貼滿瓶壁,就可以傳代了。(貼壁細胞)將培養(yǎng)瓶里的培養(yǎng)基倒去,加3-5ml(以能覆蓋細胞生長面為準)PBS或Hanks’液洗滌后棄去。加0.5-1ml 0.25%含EDTA的胰酶消化,消化時間以具體細胞為準,一般1-3分鐘,不超過5分鐘??梢苑湃?7℃培養(yǎng)箱消化。輕輕晃動瓶壁,見細胞脫落下來,加入3-5ml培養(yǎng)基終止消化。用移液管輕輕吹打瓶壁上的細胞,使之*脫落,然后將溶液吸入離心管內(nèi)離心,1000rpm/5min。棄上清,視細胞數(shù)量決定分瓶數(shù),一般一傳二,如細胞量多可一傳三,有些細胞不易傳得過稀,有些生長較快的細胞則可以多傳幾瓶,以具體細胞和經(jīng)驗為準。(懸浮細胞)用移液管輕輕吹打瓶壁,直接將溶液吸入離心管離心即可。
6,貼壁細胞 ,懸浮細胞。嚴格無菌操作。換液時,換新的細胞培養(yǎng)瓶和換新鮮的培養(yǎng)液,37度 5%CO2 培養(yǎng)
7. 收到細胞后,請鏡下觀察細胞,用恰當方式處理細胞。若懸浮的細胞較多,請離心收集細胞,接種到一個新的培養(yǎng)瓶中。棄掉原液,使用新鮮配制的培養(yǎng)基,使用進口胎牛血清. 剛接到細胞,若細胞不多時 血清濃度可以加到15%去培養(yǎng)。若細胞迏到80%左右 ,血清濃度還是在10%。
培養(yǎng)基參考
500ml基礎(chǔ)培養(yǎng)基(含:必需與非必需氨基酸、維他命、有機與無機成分、激素、生長因子與微量元素)、細胞生長添加劑、抗生素/抗菌素溶液或青霉素/鏈霉素(P/S)溶液、FBS,適合在飽和濕度為5%CO2與95%空氣的培養(yǎng)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
磷酸酶抑制劑復合物 III 10X
Bacterial Activity-Keeping Lysis Buffer細菌活性化蛋白制備裂解液
Menbrane Protein Dissolution Buffer膜蛋白溶解液
真菌和酵母用蛋白酶抑制劑復合物 100X
細菌用蛋白酶抑制劑復合物 100X
Tissue Total Protein Lysis Buffer組織蛋白裂解液
5x蛋白印跡膜再生液(強)
Inclusion body Purgation buffer包涵體洗滌液
Western and IP Cell lysis BufferWestern及IP細胞裂解液
5x蛋白印跡膜再生液(溫和)
2D Sample Extraction Buffers I(major components: urea,NP-40 )2D電泳樣品提取液 I
EZ-Buffers 10xHigh Molecules Western Transfer Buffer (no methanol)緩沖液
無蛋白封閉液 2% 封閉試劑
2D Sample Extraction Buffers II(major components: urea,CHAPS )2D電泳樣品提取液 II
2D Sample Extraction Buffers V(major components: urea,thiourea, CHAPS )2D電泳樣品提取液 V
2D Sample Extraction Buffers III(major components: urea,thiourea, CHAPS, ASB-16 )2D電泳樣品提取液 III
2D Sample Extraction Buffers IV(major components :urea,thiourea ,CHAPS, SB3-10 )2D電泳樣品提取液 IV
2D Sample Extraction Buffers VI(major components: urea,thiourea, CHAPS, ND SB201 )2D電泳樣品提取液 VI
EZ-Buffers B 10XTBS Buffer緩沖液
EZ-Buffers C 10Xwestern transfer buffer緩沖液
無蛋白封閉液 1%封閉試劑in 1XTBS
EZ-Buffers E 10XPBST buffer with Tween緩沖液
EZ-Buffers D 10XPBS buffer solution緩沖液
EZ-Buffers I 2x Primary/secondary antibody dilution buffer soluition (in PBS)緩沖液
EZ-Buffers J 2xWestern block buffer in PBS with non-fat milk緩沖液
EL-P-NPP Chromogenic Reagent kit
EZ-Buffers K 5xAP( Alkaline phosphatase) buffer solution 緩沖液
EL-OPD Chromogenic Reagent kit
EL-TMB顯色試劑
EL- OT Chromogenic Reagent kit
EL- ABTS Chromogenic Reagent kit
EZ-Buffers H 10XTBST buffer solution緩沖液
Protein Stains C: Quick& Sensitive Reversible Stain
Protein Stains A: Rapid Coomassie Stain Solution快速考馬斯亮藍染色試劑
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)緩沖液
SefinoseTM 4B 瓊脂糖凝膠
SefinoseTM 6B 瓊脂糖凝膠
SefinoseTM CL-4B 瓊脂糖凝膠
SefinoseTM CL-4B 瓊脂糖凝膠
SefinoseTM CL-6B瓊脂糖凝膠
SefinoseTM CL-6B瓊脂糖凝膠
SefinoseTM 4 Fast Flow 瓊脂糖凝膠
SefinoseTM 4 Fast Flow 瓊脂糖凝膠
SefinoseTM 6 Fast Flow瓊脂糖凝膠
SefinoseTM 6 Fast Flow瓊脂糖凝膠
Q SefinoseTM FF瓊脂糖凝膠
Q SefinoseTM FF瓊脂糖凝膠
DEAE SefinoseTM瓊脂糖凝膠
DEAE SefinoseTM瓊脂糖凝膠
SP SefinoseTM FF瓊脂糖凝膠
SP SefinoseTM FF瓊脂糖凝膠
CM SefinoseTM瓊脂糖凝膠
CM SefinoseTM瓊脂糖凝膠
Q SefinoseTM瓊脂糖凝膠
Q SefinoseTM瓊脂糖凝膠
SP SefinoseTM瓊脂糖凝膠
SP SefinoseTM瓊脂糖凝膠
Q SefinoseTM HP瓊脂糖凝膠
Q SefinoseTM HP瓊脂糖凝膠
SP SefinoseTM HP瓊脂糖凝膠
SP SefinoseTM HP瓊脂糖凝膠
Butyl SefinoseTM 4 Fast Flow丁基疏水色譜柱料
Butyl SefinoseTM 4 Fast Flow丁基疏水色譜柱料
Phenyl Sefinose 6 Fast Flow (low sub,LS) 標準載量苯基疏水色譜柱料
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