PurificationofrecombinantsBRFM166L
Induction of BRF in bacteria and purification on Ni-NTA agaroseTransform BRF plasmid into strain BL21 DE3 (pLysS) or JM25 (DE3 containing the Arg tRNA over expression plasmid under lac iQ control Kanomycin resistant)Inoculate 6 ml of a saturated overnight culture of the transformed bacteria into 2X 1 Liter of YT Amp (50 ug/ml) (and Kan (30 ug/ml) if using strain JM25) (adding 1-2 drops of antifoam A before autoc......阅读全文
Purification-of-recombinant-sBRF-M166L
Induction of BRF in bacteria and purification on Ni-NTA agaroseTransform BRF plasmid into strain BL21 DE3 (pLysS) or JM25 (DE3 containing the Arg tRNA
Antibody-Purification
This protocol includes an ammonium sulfate cut, affigel blue chromatography and affinity chromatography.1. Solutions(1) Affigel Blue Prewash0.1 M acet
Purification-of-Demethylated-Sphingomyelin
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
Protein-A-Purification-of-Antibody
1. Reagents(1) Affi-gel Protein-A Agarose (BioRad #153-6153)(2) MAPS II Binding Buffer (BioRad # 153-6161)(3) 0.314 g/ml diH2O(4) MAPS II Elution Buff
Protein-purification;-actin
Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa, 374/375 amino acids; various
Purification-of-mAb-(IgG)
1. Materials(1) Antibody 7E3, 2L sup grown in flasks, frozen and thawed overnight.(2) BioRad Affi-Gel Protein A MAPS II Buffers cat. #1530-6160 ($161.
蛋白质磷酸化
Tyrosine Kinase Assay Using Synthetic Peptides (T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assay
Purification-of-GST-Fused-Proteins
Day 1Set up an overnight culture in 100 ml LMM broth or 100 ml terrific broth containing 100ul 100 mg/mlAmpDay 2Add 40-50 ml o/n culture to 1 lt terri
Antigen-Affinity-Purification-of-Antibodies
实验概要To acquire purified antibodies (This method typically yields >95% pure specific antibodies ).实验原理 Cytokines are signaling proteins necessary for
Protein-Expression-and-Purification-Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Column-Purification-of-Demethylated-Sphingomyelin
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 colu
Protein-G-Purification-of-Antibodies
1. Reagent and Materials(1) Hi-Trap Protein G Column (Pharmacia Biotech #17-0404-01)(2) 20 mM Sodium Phosphate Buffer, pH 7.01.084 g NaH2PO4, anhydrou
Comparative-assessment-of-glycosylation-of-recombinant-human-...(六)
(22) Wu, S. W.; Pu, T. H.; Viner, R.; Khoo, K. H. Novel LC-MS/MS product dependent parallel data acquisition function and data analysis workflow f
Production-of-Recombinant-Proteins-in-SuspensionCultured-Plant-Cells
Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, toda
Comparative-assessment-of-glycosylation-of-recombinant-human-...(七)
Figure 3 MS2 spectra of 2-AB-labeled N-glycan structures. Diagnostic ions are marked with corresponding fragment structures. (a) NeuGc1NeuAc1HexNA
Comparative-assessment-of-glycosylation-of-recombinant-human-...(五)
Supporting Information(1) RP-UPLC-Q-TOF analysis of hFSHs subunits; (2) molecular weights and possible glycan structures of hFSHs subunits; (3) pr
Comparative-assessment-of-glycosylation-of-recombinant-human-...(二)
Experimental sectionChemicals and reagentsOne lot of PuregonR-HP of 50 IU/0.5mL and two lots of PuregonR-HP of 100 IU/0.5mL (rhFSH) (Organon, Oss, N
Comparative-assessment-of-glycosylation-of-recombinant-human-...(三)
LC-MS data processingN-glycan data were processed using UNIFI 1.7 with Glycobase 3+ (Waters Corporation, Milford, MA) for N-glycan structure. The pe
Comparative-assessment-of-glycosylation-of-recombinant-human-...(一)
Comparative assessment of glycosylation of recombinant human FSH and highly purified FSHHong Wang, Xi Chen, Xiaoxi Zhang, Wei Zhang, Yan Li, Hongrui Y
Comparative-assessment-of-glycosylation-of-recombinant-human-...(四)
Site-specific characterization of N-glycansFor intact N-glycopeptide analysis, chymotryptic digests of both hFSHs were subjected under UPLC equipped
DNA-Purification-from-Agarose-Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Purification-of-human-mononuclear-cells-and-neutrophils
PurposeMaterials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS4
Characterization-and-Applications-of-PlantDerived-Recombinant-Antibodies
Expression of foreign proteins in plants has become a standard technique in plant molecular biology. Various plant species have been used to produ
重组质粒(dna-recombinant-plasmid)的连接
质粒具有稳定可靠和操作简便的优点。如果要克隆较小的DNA 片段( <10kb) 且结构简单,质粒要比其它任何载体都要好。在质粒载体上进行克隆,从原理上说是很简单的,先用限制性内切酶切割质粒DNA 和目的DNA 片段,然后体外使两者相连接,再用所得到重组质粒转化细菌,即可完成。但在实际工作中,
Purification-of-acidic-phosphatase-from-mustard-seedlings
Purification of acidic phosphatase from mustard seedlings Phosphate esters are widely distributed in any organism. Nucleic acids, metabolic intermedia
Purification-of-poly(A)+-RNA-fromnb
0. (Optional)Before using Oligotex-dT30, I recommend to wash Oligotex-dT30 to remove fine particle of latex. To wash the latex, transfer appropriate a
troponin蛋白纯化-Protein-purification:-troponins
Overview TROPONINS The calcium-dependent regulatory protein complex located on the thin actin filaments of muscle comprises of TnC (17.8 kDa), TnI (2
Purification-of-Antiserum-or-Ascites-by-Protein-A/G-Chromatography
1、Required Materials and Equipment(1) Protein A or G agarose gel column (10 ml or 5 ml of packed beads; see guidelines below for choice of protein A o
DNA-Purification-from-Blood-or-Body-Fluids
实验概要试剂盒,操作简单,稍有点贵,还可以接受。实验材料blood实验步骤Protocol: DNA Purification from Blood or Body Fluids(Spin Protocol)This protocol is for purification of total (ge
RNA-Collection--Purification-from-fibrous-tissue
实验概要Provides an easy and fast method for isolating total RNA from fibrous tissues which contains contractile proteins, connective tissue and colla