Recombinant production of active microbial transglutaminase in E. coli by using self-cleavable zymogen with mutated propeptide

Microbial transglutaminase from Streptomyces mobaraensis (MTG) has been extensively utilized in meals business and in addition in analysis and medical functions, since it may possibly site-specifically modify proteins by the cross-linking response of glutamine residue and the first amino group. The recombinant expression system of MTG in E. coli supplies higher accessibility for the researchers and thus can promote additional utilization of MTG. Herein, we report manufacturing of lively and soluble MTG in E. coli through the use of a chimeric protein of tobacco etch virus (TEV) protease and MTG zymogen.

A chimera of TEV protease and MTG zymogen with native propeptide resulted in lively MTG contaminated with cleaved propeptide because of the robust interplay between the propeptide and catalytic area of MTG. Introduction of mutations of K9R and Y11A to the propeptide facilitated dissociation of the cleaved propeptide from the catalytic area of MTG and lively MTG with none contamination of the propeptide was obtained. The particular exercise of the lively MTG was 22.7±2.6 U/mg. The profitable expression and purification of lively MTG through the use of the chimera protein of TEV protease and MTG zymogen with mutations within the propeptide can advance using MTG and the researches utilizing MTG mediated cross-linking reactions.

Technology of novel trimeric fragments of human SP-A and SP-D after recombinant soluble expression in Ecoli

Surfactant therapy for neonatal respiratory misery syndrome has dramatically improved survival of preterm infants. Nonetheless, this has resulted in a markedly elevated incidence of sequelae corresponding to neonatal persistent inflammatory lung illness. The present surfactant preparations in medical use lack the pure lung defence proteins surfactant proteins (SP)-A and D. These are identified to have anti-inflammatory and anti-infective properties important for sustaining wholesome non-inflamed lungs.

Supplementation of at the moment accessible animal derived surfactant therapeutics with these anti-inflammatory proteins within the first few days of life may stop the event of inflammatory lung illness in untimely infants. Nonetheless, present methods for manufacturing of recombinant variations of SP-A and SP-D require a posh solubilisation and refolding protocol limiting expression at scale for drug improvement. Utilizing a novel solubility tag, we describe the expression and purification of recombinant fragments of human (rfh) SP-A and SP-D utilizing Escherichia coli with out the necessity for refolding.

We obtained a imply (± SD) of 23.3 (± 5.4) mg and 86 mg (± 3.5) per litre yield of rfhSP-A and rfhSP-D, respectively. rfhSP-D was trimeric and 68% sure to a ManNAc-affinity column, giving a closing yield of 57.5 mg/litre of extremely pure protein, considerably increased than the three.Three mg/litre obtained via the usual refolding protocol. Additional optimisation of this novel lab primarily based methodology may doubtlessly make rfhSP-A and rfhSP-D manufacturing extra commercially possible to allow improvement of novel therapeutics for the therapy of lung an infection and irritation.

Recombinant production of active microbial transglutaminase in E. coli by using self-cleavable zymogen with mutated propeptide

Expression, solubilization, refolding and closing purification of recombinant proteins as expressed within the type of “classical inclusion our bodies” in Ecoli

Escherichia coli has been most generally used for manufacturing of the recombinant proteins. Over-expression of the recombinant proteins is the mainspring of the inclusion our bodies formation. The refolding of those proteins into bioactive types is cumbersome and partly time-consuming. Within the current examine, we reviewed and mentioned most points concerning the restoration of "classical inclusion our bodies" by specializing in our earlier experiences.

Performing correct strategies of expression, solubilization, refolding and closing purification of those proteins, would make it attainable to get well increased quantities of pro-teins into the native type with applicable conformation. Usually, offering gentle circumstances and correct refolding buffers, would result in get well greater than 40% of inclusion our bodies into bioactive and native conformation.

Efficient Course of Growth for Excessive-level Manufacturing, Purification, Formulation, and Characterization of Recombinant Mecasermin in Ecoli

Over-production recombinant Mecasermin was achieved by investigation impact of three components, temperature, inducer quantity and tradition media in three ranges in accordance with Taguchi statistical design in E. coli in a bench-scale bioreactor. In optimum circumstances (induction temperature 28°C, TB+G medium, with 0.1 mM IPTG as inducer) 0.84 g/L Mecasermin with expression ranges of 38% of whole protein and 4.13 g/L closing dry cell biomass was produced, that is likely one of the highest values recombinant protein has been reported within the batch system.

The cell disruption was executed by lysozyme pretreatment with sonication to the environment friendly purification of Mecasermin. The remoted and washed inclusion our bodies had been solubilized in Gdn-HCl at pH 5.Four and folded with glutathione and purified with gel filtration. The purified rhIGF-1 (Mecasermin) was formulated with arginine.

Pig Connective Tissue Growth Factor (CTGF) CLIA Kit

EKU09526-96T 96T
EUR 1142.6

Pig Connective Tissue Growth Factor (CTGF)CLIA Kit

SCA010Po-10x96wellstestplate 10x96-wells test plate
EUR 8096.83
Description: This is Double-antibody Sandwich Chemiluminescent immunoassay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF)CLIA Kit

SCA010Po-1x48wellstestplate 1x48-wells test plate
EUR 771.97
Description: This is Double-antibody Sandwich Chemiluminescent immunoassay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF)CLIA Kit

SCA010Po-1x96wellstestplate 1x96-wells test plate
EUR 1051.39
Description: This is Double-antibody Sandwich Chemiluminescent immunoassay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF)CLIA Kit

SCA010Po-5x96wellstestplate 5x96-wells test plate
EUR 4371.26
Description: This is Double-antibody Sandwich Chemiluminescent immunoassay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF) CLIA Kit

4-SCA010Po
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  • 10 plates of 96 wells
  • 5 plates of 96 wells
  • 1 plate of 96 wells
Description: Double-antibody Sandwich chemiluminescent immunoassay for detection of Pig Connective Tissue Growth Factor (CTGF)Serum, plasma and other biological fluids

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

abx573322-96tests 96 tests
EUR 990

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

20-abx154906
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  • 10 × 96 tests
  • 5 × 96 tests
  • 96 tests

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKN44549-48T 48T
EUR 409.22

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKN44549-5x96T 5x96T
EUR 2776.85

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKN44549-96T 96T
EUR 584.6

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKU03438-48T 48T
EUR 600.25

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKU03438-5x96T 5x96T
EUR 4073.13

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

EKU03438-96T 96T
EUR 857.5

Pig Connective tissue growth factor (CTGF) ELISA Kit

RK09315 96T
EUR 280

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

SEA010Po-10x96wellstestplate 10x96-wells test plate
EUR 6117.62
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma, tissue homogenates and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

SEA010Po-1x48wellstestplate 1x48-wells test plate
EUR 608.98
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma, tissue homogenates and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

SEA010Po-1x96wellstestplate 1x96-wells test plate
EUR 818.54
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma, tissue homogenates and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

SEA010Po-5x96wellstestplate 5x96-wells test plate
EUR 3323.45
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Pig Connective Tissue Growth Factor (CTGF) in serum, plasma, tissue homogenates and other biological fluids.

Pig Connective Tissue Growth Factor (CTGF) ELISA Kit

4-SEA010Po
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  • 10 plates of 96 wells
  • 5 plates of 96 wells
  • 1 plate of 96 wells
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Pig Connective Tissue Growth Factor (CTGF) in samples from Serum, plasma, tissue homogenates and other biological fluids. with no significant corss-reactivity with analogues from other species.

Pig CTGF/Connective tissue growth factor ELISA Kit

EK19168 96Т
EUR 650

Pig CTGF(Connective Tissue Growth Factor) ELISA Kit

ELK5960-48T 48T Ask for price
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Pig CTGF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Pig CTGF. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Pig CTGF, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Pig CTGF in the samples is then determined by comparing the OD of the samples to the standard curve.

Pig CTGF(Connective Tissue Growth Factor) ELISA Kit

ELK5960-96T 96T Ask for price
Description: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Pig CTGF. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Pig CTGF. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Pig CTGF, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Pig CTGF in the samples is then determined by comparing the OD of the samples to the standard curve.

Human Connective tissue growth factor (CTGF)

1-CSB-EP006147HU
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  • 100ug
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  • 1MG
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  • 50ug
Description: Recombinant Human Connective tissue growth factor(CTGF) expressed in E.coli

Human Connective tissue growth factor (CTGF)

1-CSB-EP006147HU1
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  • 100ug
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  • 1MG
  • 200ug
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  • 50ug
Description: Recombinant Human Connective tissue growth factor(CTGF),partial expressed in E.coli

Mouse Connective tissue growth factor (Ctgf)

1-CSB-EP006147MO
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  • 100ug
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  • 1MG
  • 200ug
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  • 50ug
Description: Recombinant Mouse Connective tissue growth factor (Ctgf),partial expressed in E.coli

Mouse Connective tissue growth factor (Ctgf)

1-CSB-EP006147MOe0
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  • 100ug
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Description: Recombinant Mouse Connective tissue growth factor(Ctgf),partial expressed in E.coli

Active Connective Tissue Growth Factor (CTGF)

APA010Ra01 10ug
EUR 182

Active Connective Tissue Growth Factor (CTGF)

4-APA010Ra01
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  • 100 ug
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  • 5 mg
Description: Recombinant Rat Connective Tissue Growth Factor expressed in: Available from E.coli, Yeast, Baculovirus and Mammalian cells

Active Connective Tissue Growth Factor (CTGF)

RPU55435-100ug 100ug
EUR 562.1

Active Connective Tissue Growth Factor (CTGF)

RPU55435-1mg 1mg
EUR 2489.5

Active Connective Tissue Growth Factor (CTGF)

RPU55435-50ug 50ug
EUR 451

Guinea pig Connective Tissue Growth Factor (CTGF) ELISA Kit

abx358103-96tests 96 tests
EUR 990

Guinea pig Connective Tissue Growth Factor (CTGF)ELISA Kit

NSL1253Gp 96T
EUR 528

Connective Tissue Growth Factor (CTGF) Antibody

20-abx109670
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103225
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103226
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103227
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103228
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103229
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103230
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx103231
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx104836
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  • 100 ug
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  • 1 mg
  • 200 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx131768
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  • 100 ug
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  • 200 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx123700
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  • 100 ul
  • 200 ul

Connective Tissue Growth Factor (CTGF) Antibody

20-abx125726
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  • 100 ul
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx125727
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  • 100 ul
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx125728
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  • 100 ul
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx132193
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx132350
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

20-abx132360
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  • 100 ug
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Connective Tissue Growth Factor (CTGF) Antibody

abx037800-100ug 100 ug
EUR 469.2

Connective Tissue Growth Factor (CTGF) Antibody

abx033675-400ul 400 ul
EUR 627.6

Connective Tissue Growth Factor (CTGF) Antibody

abx033675-80l 80 µl
EUR 343.2

Connective Tissue Growth Factor (CTGF) Antibody

abx033676-400ul 400 ul
EUR 627.6

Connective Tissue Growth Factor (CTGF) Antibody

abx033676-80l 80 µl
EUR 343.2

Connective Tissue Growth Factor (CTGF) Antibody

abx027387-400ul 400 ul
EUR 627.6

Connective Tissue Growth Factor (CTGF) Antibody

abx027387-80l 80 µl
EUR 343.2

Connective Tissue Growth Factor (CTGF) Antibody

abx025473-100ul 100 ul
EUR 627.6

Connective tissue growth factor (CTGF) Antibody

20-abx214776
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  • 100 ul
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Connective Tissue Growth Factor (CTGF) Antibody

abx232053-100ug 100 ug
EUR 610.8

Connective Tissue Growth Factor (CTGF) Antibody

abx232054-100ug 100 ug
EUR 610.8

Connective Tissue Growth Factor (CTGF) Antibody

20-abx171909
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  • 1 mg
  • 200 ug

Connective Tissue Growth Factor (CTGF) Antibody

20-abx171910
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  • 1 mg
  • 200 ug

Connective Tissue Growth Factor (CTGF) Antibody

20-abx225128
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  • 100 ul
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  • 50 ul

Connective Tissue Growth Factor (CTGF) Antibody

20-abx175995
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  • 1 mg
  • 200 ug

Connective Tissue Growth Factor (CTGF) Antibody

20-abx175996
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  • 1 mg
  • 200 ug

Connective Tissue Growth Factor (CTGF) Antibody

20-abx175997
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  • 1 mg
  • 200 ug

Connective Tissue Growth Factor (CTGF) Antibody

abx433502-200ul 200 ul
EUR 543.6

Connective Tissue Growth Factor (Ctgf) Antibody

20-abx319302
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  • 100 ug
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ELISA kit for Pig CTGF (Connective Tissue Growth Factor)

ELK5960 1 plate of 96 wells
EUR 631.2
Description: A sandwich ELISA kit for detection of Connective Tissue Growth Factor from Pig in samples from blood, serum, plasma, cell culture fluid and other biological fluids.

CTGF (untagged)-Human connective tissue growth factor (CTGF)

SC118973 10 µg Ask for price

Connective Tissue Growth Factor (CTGF) Polyclonal Antibody (Pig)

4-PAA010Po01
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  • 100ul
  • 10ml
  • 1ml
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  • 20ul
Description: A Rabbit polyclonal antibody against Pig Connective Tissue Growth Factor (CTGF)

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Bo01 10ug
EUR 192

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Bo01
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  • 100 ug
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  • 1 mg
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  • 50ug
  • 5 mg
Description: Recombinant Bovine Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Cp01 10ug
EUR 159

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Ga01 10ug
EUR 164

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Hu01 10ug
EUR 168

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Hu01
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  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Human Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Hu02 10ug
EUR 168

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Hu02
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  • 100 ug
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  • 1 mg
  • 200 ug
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  • 50ug
  • 5 mg
Description: Recombinant Human Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Hu03 10ug
EUR 172

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Hu04 10ug
EUR 170

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Mu01 10ug
EUR 180

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Mu01
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  • 100 ug
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  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Mouse Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Ov01 10ug
EUR 204

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Ov01
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  • 100 ug
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  • 1 mg
  • 200 ug
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  • 50ug
  • 5 mg
Description: Recombinant Sheep Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Po01 10ug
EUR 192

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Po01
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  • 100 ug
  • 10ug
  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Pig Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Ra01 10ug
EUR 140

Recombinant Connective Tissue Growth Factor (CTGF)

4-RPA010Ra01
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  • 100 ug
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  • 1 mg
  • 200 ug
  • 500 ug
  • 50ug
  • 5 mg
Description: Recombinant Rat Connective Tissue Growth Factor expressed in: E.coli

Recombinant Connective Tissue Growth Factor (CTGF)

RPA010Rb01 10ug
EUR 196

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58387-100ug 100ug
EUR 551.1

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58387-1mg 1mg
EUR 2558.4

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58387-50ug 50ug
EUR 428.5

Recombinant Connective Tissue Growth Factor (CTGF)

RPU56456-100ug 100ug
EUR 414.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU56456-1mg 1mg
EUR 1922

Recombinant Connective Tissue Growth Factor (CTGF)

RPU56456-50ug 50ug
EUR 295.7

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58764-100ug 100ug
EUR 658.6

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58764-1mg 1mg
EUR 3057.6

Recombinant Connective Tissue Growth Factor (CTGF)

RPU58764-50ug 50ug
EUR 485.1

Recombinant Connective Tissue Growth Factor (CTGF)

RPU57939-100ug 100ug
EUR 535.1

Recombinant Connective Tissue Growth Factor (CTGF)

RPU57939-1mg 1mg
EUR 2483.5

Recombinant Connective Tissue Growth Factor (CTGF)

RPU57939-50ug 50ug
EUR 415.9

Recombinant Connective Tissue Growth Factor (CTGF)

RPU54283-100ug 100ug
EUR 591.8

Recombinant Connective Tissue Growth Factor (CTGF)

RPU54283-1mg 1mg
EUR 2620.8

Recombinant Connective Tissue Growth Factor (CTGF)

RPU54283-50ug 50ug
EUR 475.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40981-100ug 100ug
EUR 517

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40981-1mg 1mg
EUR 2293.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40981-50ug 50ug
EUR 415.8

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40982-100ug 100ug
EUR 554.4

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40982-1mg 1mg
EUR 2457

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40982-50ug 50ug
EUR 445.5

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40983-100ug 100ug
EUR 591.8

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40983-1mg 1mg
EUR 2620.8

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40983-50ug 50ug
EUR 475.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40984-100ug 100ug
EUR 431.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40984-1mg 1mg
EUR 1911

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40984-50ug 50ug
EUR 346.5

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40985-100ug 100ug
EUR 517

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40985-1mg 1mg
EUR 2293.2

Recombinant Connective Tissue Growth Factor (CTGF)

RPU40985-50ug 50ug
EUR 415.8

Mecasermin protein saved stability at 4°C for as much as 2 years. The quantitative and qualitative management indicated that Mecasermin is expressed appropriately (with out the preliminary methionine by Mass spectrometry), pure (with out endotoxin and different protein impurities), appropriate folding (FTIR, RF-HPLC), monomer type (SEC-HPLC) and lively (bioactivity check). Additionally, the purification outcomes revealed that expression in low temperature outcomes the environment friendly purification of the over-produced Mecasermin with a excessive amount and high quality.

 

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