蛋白质技术

lution of GST-Fusion Protein from Glutathione Agarose1. Wash the GST-fusion protein glutathione-agarose complex once with 1% Triton X-100 in PBS.2. Resuspend the complex in 1% Triton X-100 in PBS.3. ...

Small Scale GST-Tag Purification Under Nature Conditions According to AMERSHAM-BIOSCIENCES-GST fusion protein Handbook (pdf)Aliquot of Cell Pellet after Induction The idea is to aliquot cells after i ...

His-tag protein purification using Ni-NTA magnetic beadsMaterials and reagentsExtraction buffer20 mM Tris-HCl pH 8.0 1 mM PMSF1 mg/ml lysozyme0.05% Triton X-100 ...

His-tag protein purification using Ni-NTA magnetic beadsMaterials and reagentsExtraction buffer20 mM Tris-HCl pH 8.0 1 mM PMSF1 mg/ml lysozyme0.05% Triton X-100 ...

Purification of Kar3 Motor Domain ProteinMaterials Induced cells (2 - 5 g pellet of pET/Kar3 in BL31(DE3)pLysS host cells) (See note #1)HEM buffer = 10 mM HEPES pH 7.21 mM MgCL21 mM DTT1 mM EGTAHEM + ...

Purification of Kar3 Motor Domain ProteinMaterials Induced cells (2 - 5 g pellet of pET/Kar3 in BL31(DE3)pLysS host cells) (See note #1)HEM buffer = 10 mM HEPES pH 7.21 mM MgCL21 mM DTT1 mM EGTAHEM + ...

Native purification of His-tagged Proteins from Sf9 cellsProtocol is for 150 ml of cell culture.1. Spin down cells and wash 1X in 10 ml ice cold PBS.2. Resuspend pellet in 20 ml lysis buffer: 50 mM Tr ...

Small scale His-Tag fusion protein purification under denaturative conditionsIntroduction High levels of expression of recombinant proteins in a bacterial system can lead to the formation of insoluble ...

Purification of KHC Motor Domain ProteinMaterials Induced cells (2 - 5 g pellet of pET/KHC in BL21(DE3)pLysS host cells)HEM buffer = 10 mM HEPES pH 7.21 mM DTT1 mM MgCL21 mM EGTAHEM + 25 mM NaCl HEM + ...

Purification of Ncd Motor Domain ProteinMaterials Induced cells (2 - 5 g pellet of pET/Ncd in BL21(DE3)pLysS host cells)HEM buffer = 10 mM HEPES pH 7.21 mM DTT1 mM MgCL21 mM EGTAHEM + 80 mM NaCl HEM + ...

Purification of Ncd Motor Domain ProteinMaterials Induced cells (2 - 5 g pellet of pET/Ncd in BL21(DE3)pLysS host cells)HEM buffer = 10 mM HEPES pH 7.21 mM DTT1 mM MgCL21 mM EGTAHEM + 80 mM NaCl HEM + ...

Protein purification; actin Overview ACTINThe most abundant muscle and non-muscle cytoskeletal protein. MW 42 kDa 374/375 amino acids; various isoforms; structure resolved at 2.8 Angstrom. 208 ...

Purification of CD21) Cells are pelleted (~3 - 4000 g for 10 min.) and resuspended in pre-chilled 50 mM malonate (pH 5.2-5.3) with 1mM EDTA (~40ml of buffer per litre of culture).2) Lyse the cells wit ...

OverviewTROPOMYOSIN An ubiquitous actin binding protein arranged in alpha-helical coiled-coil of 410 Angstrom length. MaterialSource: rabbit skeletal muscle Equipment: • hydroxylapatite column ...

Purification of ICAM-1by Mazen Ferzly 06/22/2000 PurposeThe purification of ICAM-1 on R6.5 anti-ICAM-1 column.Materials Triethylamine NaCl Octyl Glucoside ( n-octyl-b-D-glucopyranoside Sigma Cat. No. ...

OverviewSKELETAL MUSCLE MYOSINA two-headed hexomeric actin-activated ATPase consisting of two heavy chains of ~200 kDa and four light chains of ~20kDa. The tail portion consists of an alpha-helical c ...

Overview TROPONINS The calcium-dependent regulatory protein complex located on the thin actin filaments of muscle comprises of TnC (17.8 kDa) TnI (20.8 kDa) and TnT (30.5 kDa). These proteins are i ...

Home-made Taq Polymerase PurificationI also have the bacteria containing the clone. It appears to produce lots of Taq and is quite stable.The proceedure takes 4 days start to (15 000 units of Taq) fin ...

. Overview This protocol provides considerations and generalized procedures for the use of dialysis to alter the composition of a protein solution. It is a useful step in adjusting a protein samp ...

Grow a 2 ml culture 24 hr. at 30oC in selective media When culture is ready use it to inoculate about 55 ml (50 ml plus 5 for O.D.600 readings) of selective media. Grow to mid-logarithmicphase (O.D.60 ...