ELISPOTProtocol
實驗概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on the single cell level. The popularity of this assay has seen resurgence in recent years as researchers attempt to gain a better understanding of immune responses in a variety of applications. The following protocol is an example of a typ......閱讀全文
ELISPOT-protocol
實驗概要The procedure ?below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits ?have been designed for detection of various cytokines and g
ELISPOT-Protocol
實驗概要The ?Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method ?of measuring the antibody or cytokine production of immune cells on t
ELISPOT-Protocol
實驗概要The ?Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method ?of measuring the antibody or cytokine production of immune cells on t
ELISPOT
實驗材料 抗原試劑、試劑盒 PBSPRMI1640培養基氯化銨儀器、耗材 PVDF膜96孔板實驗步驟 1. PBS 溶解抗原為30 μg/ml,加100 μl/孔于PVDF 膜鋪底的96 孔滅菌板過夜;?2. 第二天吸去包被液后,加5%FCS 的PRMI 1640 培養基100 μl 封閉1 小時,
ELISPOT
隨著酶聯免疫分析技術在醫學及生物學領域的廣泛應用,使體外檢測各種細胞因子及抗體研究有了新的突破。在研究免疫應答機制時以往常用用酶聯免疫吸附法(ELISA)檢測體液中游離的細胞因子(CK)或抗體,但由于游離的循環抗體或CK的半哀期不同,使之在體液中不斷的被代謝或與靶器官結合,而不能確切的反映體內的抗體
The-Impact-of-Harmo...-(一)
實驗概要During more than ?25 years of application in immunological sciences, ELISPOT has been ?established as a routine, robust, versatile, and reliable a
Dual-ELISPOT
實驗概要We provide a ?protocol using an FITC-conjugated primary antibody and a biotinylated ?primary antibody. Those are in turn recognized by anti-FITC H
The-Impact-of-Harmo...-(二)
In ?addition, proficiency panel projects have demonstrated that even after ?an experiment was done and spots were counted, considerable variation ?can
ELISpot實驗操作流程
ELISPOT全名為酶聯免疫斑點檢測,英文:Enzyme-linked Immunospot Assay。它結合了細胞培養技術與酶聯免疫吸附技術(即ELISA技術),能夠在單細胞水平檢測細胞因子的分泌情況。其技術原理,一句話概括就是:用抗體捕獲培養中的細胞分泌的細胞因子,并以酶聯斑點顯色的方式將其表
NAi-protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
Immunoblot-Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
RNAi-protocol
?siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
PCR-protocol
PCR reactionProtocol for 50μl reaction - adjust amounts if necessary, for a 20μl reaction use the same volumes of primer and dNTP-mix, but adjust the
Immunoprecipitation-Protocol
實驗概要Immunoprecipitation ?is a procedure by which proteins or peptides that react specifically ?with an antibody are removed from solution and examined
ELISA-protocol
ELISA protocol:1.取5-10ul BMMY表達上清用0.05M NaHCO3稀釋到100ul鋪ELISA板,37度或室溫振蕩大于1小時。注意一定要做一個GS115空菌株表達上清作為陰性對照,最好還找一個帶有histag的蛋白作為陽性對照。2.TPBS洗板3次,方法:倒掉鋪板液,倒置于
RLGS-protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.????????
ELISPOT-(Enzymelinked-ImmunoSPOT)
IntroductionEnzyme-linked immunosorbent spot (ELISPOT) assays were originally developed to enumerate B cells secreting antigen-specific antibodies, bu
ELISPOT與ELISA的區別
1.ELISA通過顯色反應,在酶標儀上測定吸光度,與標準曲線比較得出可溶性蛋白總量。2.ELISPOT通過顯色反應,在細胞分泌這種可溶性蛋白的相應位置上顯現清晰可辨的斑點,可直接在顯微鏡下或通過儀器對斑點進行計數,1個斑點代表1個細胞,從而計算出分泌該蛋白的細胞的頻率。3.由于是單細胞水平檢測,EL
酶聯免疫斑點實驗(ELIspot)——酶聯免疫斑點實驗(ELIspot)
實驗材料血液樣品細胞樣品試劑、試劑盒70%乙醇PBS脫脂奶粉鏈霉親和素檢測抗體包被抗體儀器、耗材PVDF膜96孔培養板硝基纖維素底板免疫斑點板酶標板實驗步驟一、包被96孔板?用70%乙醇浸潤96孔板中的PVDF膜30 s?加入捕獲抗體(PBS稀釋),4℃過夜?倒空板中包被液,輕輕在紙上拍干,用PBS
ELISPOT(酶聯免疫斑點法)技術
酶聯免疫斑點技術(Enzyme Linked Immunospot Assay, ELISPOT)是細胞免疫學研究中最敏感的檢測方法之一,可以在單細胞水平對抗體分泌細胞(ASC)及細胞因子(CK) 分泌細胞進行檢測。由于是單細胞水平檢測,ELISPOT比ELISA 和有限稀釋法等具有更高的靈敏性,能
酶聯免疫斑點實驗(ELIspot)_酶聯免疫斑點實驗(ELIspot)2
實驗方法原理ELISPOT技術原理,一句話概括就是:用抗體捕獲培養中的細胞分泌的細胞因子,并以酶聯斑點顯色的方式將其表現出來。實驗材料細胞試劑、試劑盒PBSPBST乙醇包被抗體封閉液抗體稀釋液檢測抗體酶聯親和素AEC顯色液PHA刺激物U-Cytech無血清培養基儀器、耗材ELISPOT讀數儀超凈工作
Transformation-Protocol-for-Arabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial sus
Microarray-Hybridization-Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
Nuclear-Extraction-Protocol
實驗概要The procedure presented below describes a method for extracting nuclear from several cell lines of human origin.主要試劑Hypotonic Buffer Solution20 mM
Immunofluorescence-Microscopy-Protocol
實驗概要Immunofluorescence ?allows the imaging of a specific factor in cells or tissue sections ?through the use of a specific antibody chemically which i
RNA-Isolation-Protocol
Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagen
Yale-Immunofluorescence-Protocol
實驗概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要試劑Reagents1.?384-well view plates (Aurora)2.?HUVEC (pooled, L
Bacteria-Culture-Protocol
Bacteria Culture ProtocolBy 徐曉政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
Intracellular-Staining-Protocol
1.?Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min
Histone-blotting-protocol
實驗概要?Western blot detection of histone proteins.?實驗步驟?The ?following protocol refers to the western blot detection of histone ?proteins derived from p