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Expression and purification of functional recombinant CUL2•RBX1 from E. coli

by Jacob
Cullin-2 (CUL2) primarily based cullin-RING ligases (CRL2s) comprise a household of ubiquitin E3 ligases that exist solely in multi-cellular organisms and are essential for mobile processes equivalent to embryogenesis and viral pathogenesis. CUL2 is the scaffold protein that binds one of the interchangeable substrate receptor modules, which consists of adaptor proteins and the substrate receptor protein. The VHL protein is a substrate receptor identified to focus on hypoxia-inducible issue α (HIF1α) for ubiquitination and degradation.
Because of its essential function within the ubiquitination of necessary mobile components equivalent to HIF1α, CRL2s have been investigated for his or her organic features and the event of novel therapeutics towards illnesses. Given the significance of CRL2s in organic and biomedical analysis, strategies that effectively produce functional CUL2 proteins will significantly facilitate research on the mechanism and regulation of CRL2s.
Here, we report two cost-effective programs for the expression and purification of recombinant human CUL2 from E. coli cells. The purified CUL2 proteins have been ~ 95% pure, may bind their substrate receptor modules, and have been enzymatically lively in transferring ubiquitin or ubiquitin-like protein to the corresponding substrate in in vitro assays. The introduced methodological developments will assist advance analysis in CRL2 perform and regulation.

Milligram scale expression, refolding, and purification of Bombyx mori cocoonase utilizing a recombinant E. coli system

Silk is one of probably the most versatile biomaterials with signature properties of excellent mechanical energy and flexibility. A possible avenue for growing extra environmentally pleasant silk manufacturing is to make use of the silk moth (Bombyx mori) cocoonase, this may on the similar time improve the likelihood for utilizing the byproduct, sericin, as a uncooked materials for different functions.
Cocoonase is a serine protease utilized by the silk moth to melt the cocoon to allow its escape after accomplished metamorphosis. Cocoonase selectively degrades the glue protein of the cocoon, sericin, with out affecting the silk-fiber made of the protein fibroin. Cocoonase might be recombinantly produced in E. coli, nevertheless, it’s solely discovered as insoluble inclusion our bodies.
To clear up this drawback and to have the ability to make the most of the advantages related to an E. coli primarily based expression system, we’ve developed a protocol that allows the manufacturing of soluble and functional protease within the milligram/liter scale. The core of the protocol is refolding of the protein in a buffer with a redox potential that’s optimized for formation of native and intramolecular di-sulfide bridges.
The redox potential was balanced with outlined concentrations of decreased and oxidized glutathione. This E coli primarily based manufacturing protocol will, along with construction dedication, additionally allow modification of cocoonase each in phrases of catalytic perform and stability. These components might be helpful parts within the growth of alternate silk manufacturing methodology.

Expression of Bacillus amyloliquefaciens transglutaminase in recombinant E. coli below the management of a bicistronic plasmid system in DO-stat fed-batch bioreactor cultivations

We studied the expression of Bacillus amyloliquefaciens transglutaminase cloned in Escherichia coli BL21(DE3)pLysS harboring the plasmid pBAD/3C/bTGase, a bicistronic expression system, in bioreactor cultivation. Batch and fed-batch managed as DO-stat methods have been employed for the manufacturing of the recombinant enzyme. In 30 h-batch cultivations utilizing Terrific broth (TB), 6 g/L of biomass and 3.12 U/mgprotein of transglutaminase exercise have been obtained.
Expression and purification of functional recombinant CUL2•RBX1 from E. coli
DO-stat fed-batch cultivations below the management of oxygen focus (DO-stat) utilizing TB as medium however fed with glucose allowed the increment in biomass formation (17.5 g/L) and enzyme exercise (6.43 U/mgprotein). DO-stat fed-batch utilizing mineral medium (M9) and fed with glucose below the identical situations produced even greater enzymatic exercise (9.14 U/mgprotein).
The pH impact was investigated, and one of the best enzymatic exercise may very well be noticed at pH 8. In all cultivations, the bicistronic system remained secure, with 100% of plasmid-bearing cells. These outcomes present that E. coli bearing bicistronic plasmid constructs to specific recombinant TGase may very well be cultivated in bioreactors below DO-stat fed-batch utilizing mineral medium and it’s a promising technique in future optimizations to provide this necessary enzyme.

Characterization of Recombinant E. coli Expressing a Novel Fucosidase from Bacillus cereus 2-Eight belonging to GH95 Family

Fucoidan oligosaccharides possesses numerous physicochemical and organic actions. Specific glycoside hydrolases are helpful instruments for degrading polysaccharides to provide oligosaccharides. In this examine, BcFucA, a novel fucosidase belonging to GH95 household from Bacillus cereus 2-8, was cloned into pET21a vector, expressed in E. coli BL21 (DE3) and characterised.

aida Antibody
CSB-PA761322XA01DIL-10mg 10mg Ask for price
Description: Recombinant Danio rerio (Zebrafish) (Brachydanio rerio) aida protein

AIDA Antibody
DF15062 100ul
EUR 420

AIDA Antibody
DF15062-100ul 100ul
EUR 168
Description: IHC,IF/ICC,ELISA(peptide)

AIDA Antibody
DF15062-200ul 200ul
EUR 210
Description: IHC,IF/ICC,ELISA(peptide)

AIDA Antibody
GWB-MW763G 50ug Ask for price

AIDA Antibody
GWB-MW764H 50ug Ask for price

AIDA Antibody
MBS7134830-005mL 0.05mL
EUR 190

AIDA Antibody
MBS7134830-01mL 0.1mL
EUR 270

AIDA Antibody
MBS7134830-5x01mL 5x0.1mL
EUR 1205

AIDA Antibody
MBS9630350-01mL 0.1mL
EUR 260

AIDA Antibody
MBS9630350-02mL 0.2mL
EUR 305

AIDA Antibody
MBS9630350-5x02mL 5x0.2mL
EUR 1220

AIDA Antibody
MBS9632733-1mg 1mg
EUR 375

AIDA Antibody
MBS9632733-5x1mg 5x1mg
EUR 1545

AIDA (untagged)-Human axin interactor, dorsalization associated (AIDA)
SC112378 10 µg Ask for price

AIDA cDNA Clone
MBS1268688-001mgPlasmid02mLGlycerolStock 0.01mgPlasmid+0.2mLGlycerol-Stock
EUR 200

AIDA cDNA Clone
MBS1268688-5x001mgPlasmid5x02mLGlycerolStock 5x0.01mgPlasmid+5x0.2mLGlycerol-Stock
EUR 855

Mouse AIDA siRNA
20-abx907083
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  • 15 nmol
  • 30 nmol

Human AIDA siRNA
20-abx907084
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  • 15 nmol
  • 30 nmol

AIDA siRNA (Human)
MBS8227846-15nmol 15nmol
EUR 405

AIDA siRNA (Human)
MBS8227846-30nmol 30nmol
EUR 565

AIDA siRNA (Human)
MBS8227846-5x30nmol 5x30nmol
EUR 2450

AIDA siRNA (Mouse)
MBS8209465-15nmol 15nmol
EUR 405

AIDA siRNA (Mouse)
MBS8209465-30nmol 30nmol
EUR 565

AIDA siRNA (Mouse)
MBS8209465-5x30nmol 5x30nmol
EUR 2450

aidA-I Antibody
CSB-PA183385XA01ENL-02mg 0.2mg Ask for price
Description: Recombinant Escherichia coli aidA-I protein

aidA-I Antibody
CSB-PA183385XA01ENL-10mg 10mg Ask for price
Description: Recombinant Escherichia coli aidA-I protein

anti- AIDA antibody
FNab00234 100µg
EUR 606.3
Description: Antibody raised against AIDA

AIDA cloning plasmid
CSB-CL839294HU-10ug 10ug
EUR 279.6
Description: A cloning plasmid for the AIDA gene.

AIDA (GFP-tagged) - Human axin interactor, dorsalization associated (AIDA)
RG217521 10 µg Ask for price

Recombinant Human AIDA
MBS7611060-005mg 0.05mg
EUR 405

Recombinant Human AIDA
MBS7611060-02mg 0.2mg
EUR 760

Recombinant Human AIDA
MBS7611060-1mg 1mg
EUR 2175

Recombinant Human AIDA
MBS7611060-5x1mg 5x1mg
EUR 8410

Recombinant Human AIDA
MBS7616425-005mg 0.05mg
EUR 470

Recombinant Human AIDA
MBS7616425-02mg 0.2mg
EUR 885

Recombinant Human AIDA
MBS7616425-1mg 1mg
EUR 2540

Recombinant Human AIDA
MBS7616425-5x1mg 5x1mg
EUR 9820

AIDA, 1-306aa, Human, His tag, E Coli
MBS205394-0004mg 0.004mg
EUR 215

AIDA, 1-306aa, Human, His tag, E Coli
MBS205394-002mg 0.02mg
EUR 340

AIDA, 1-306aa, Human, His tag, E Coli
MBS205394-01mg 0.1mg
EUR 765

AIDA, 1-306aa, Human, His tag, E Coli
MBS205394-5x01mg 5x0.1mg
EUR 3195

Aida (Myc-DDK-tagged) - Mouse axin interactor, dorsalization associated (Aida)
MR204280 10 µg Ask for price

AIDA (Myc-DDK-tagged)-Human axin interactor, dorsalization associated (AIDA)
RC217521 10 µg Ask for price

Aida (NM_181732) Mouse Recombinant Protein
E45M47447M5 20 ug
EUR 1924.54

AIDA protein (His tag)
80R-2681 20 ug
EUR 329
Description: Purified recombinant AIDA protein (His tag)

AIDA ELISA KIT|Human
EF007672 96 Tests
EUR 826.8

Mouse Aida ELISA KIT
ELI-24374m 96 Tests
EUR 1038

Human AIDA ELISA KIT
ELI-11961h 96 Tests
EUR 988.8

AIDA Antibody (C-term)
E45R04233G-1 100 ul
EUR 395

AIDA Antibody (C-term)
E45R04233G-4 50 ul
EUR 295

AIDA Antibody (C-term)
MBS9201306-008mL 0.08mL
EUR 210

AIDA Antibody (C-term)
MBS9201306-04mL 0.4mL
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MBS9201306-5x04mL 5x0.4mL
EUR 1910

Mouse AIDA shRNA Plasmid
20-abx979944
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  • 150 µg
  • 300 µg

Human AIDA shRNA Plasmid
20-abx962119
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  • 150 µg
  • 300 µg

Human AIDA Protein Lysate
MBS137038-002mg 0.02mg
EUR 365

Human AIDA Protein Lysate
MBS137038-5x002mg 5x0.02mg
EUR 1410

AIDA Recombinant Rabbit mAb
BS48755 50 ul
EUR 398
Description: Store at -20°C. Supplied in 50mM Tris-Glycine(pH 7.4), 0.15M NaCl, 40%Glycerol, 0.01% sodium azide and 0.05% BSA. Stable for 12 months from date of receipt.

AIDA Antibody, HRP Conjugated
MBS7134831-005mL 0.05mL
EUR 190

AIDA Antibody, HRP Conjugated
MBS7134831-01mL 0.1mL
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AIDA Antibody, HRP Conjugated
MBS7134831-5x01mL 5x0.1mL
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AIDA Antibody, FITC Conjugated
MBS7134832-005mL 0.05mL
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AIDA Antibody, FITC Conjugated
MBS7134832-01mL 0.1mL
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AIDA Antibody, FITC Conjugated
MBS7134832-5x01mL 5x0.1mL
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Recombinant Human AIDA Protein
E30P62280A 50 ug
EUR 295

AIDA Recombinant Protein (Human)
RP000832 100 ug Ask for price

AIDA Recombinant Protein (Mouse)
RP115019 100 ug Ask for price

AIDA Rabbit Polyclonal Antibody
E10G31837 100 μl
EUR 275
Description: Biotin-Conjugated, FITC-Conjugated , AF350 Conjugated , AF405M-Conjugated ,AF488-Conjugated, AF514-Conjugated ,AF532-Conjugated, AF555-Conjugated ,AF568-Conjugated , HRP-Conjugated, AF405S-Conjugated, AF405L-Conjugated , AF546-Conjugated, AF594-Conjugated , AF610-Conjugated, AF635-Conjugated , AF647-Conjugated , AF680-Conjugated , AF700-Conjugated , AF750-Conjugated , AF790-Conjugated , APC-Conjugated , PE-Conjugated , Cy3-Conjugated , Cy5-Conjugated , Cy5.5-Conjugated , Cy7-Conjugated Antibody

AIDA Antibody, Biotin Conjugated
MBS7134833-005mL 0.05mL
EUR 190

AIDA Antibody, Biotin Conjugated
MBS7134833-01mL 0.1mL
EUR 270

AIDA Antibody, Biotin Conjugated
MBS7134833-5x01mL 5x0.1mL
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AIDA sgRNA CRISPR Lentivector (Human) (Target 1)
K0062102 1.0 ug DNA
EUR 184.8

Aida sgRNA CRISPR Lentivector (Mouse) (Target 1)
K4069402 1.0 ug DNA
EUR 184.8

C1orf80 (AIDA) (NM_022831) Human Recombinant Protein
E45H35418M5 20 ug
EUR 785.42

C1orf80 (AIDA) (NM_022831) Human Recombinant Protein
PROTQ96BJ3 Regular: 10ug
EUR 1249
Description: Recombinant protein of human axin interactor, dorsalization associated (AIDA)

AIDA (C-term) Rabbit pAb
E2613568 100ul
EUR 225
Description: Available in various conjugation types.

AIDA (C-Term) Rabbit pAb
MBS8552977-01mL 0.1mL
EUR 305

AIDA (C-Term) Rabbit pAb
MBS8552977-01mLAF405L 0.1mL(AF405L)
EUR 565

AIDA (C-Term) Rabbit pAb
MBS8552977-01mLAF405S 0.1mL(AF405S)
EUR 565

AIDA (C-Term) Rabbit pAb
MBS8552977-01mLAF610 0.1mL(AF610)
EUR 565

AIDA (C-Term) Rabbit pAb
MBS8552977-01mLAF635 0.1mL(AF635)
EUR 565

AIDA ORF Vector (Human) (pORF)
ORF000278 1.0 ug DNA
EUR 114

Aida ORF Vector (Mouse) (pORF)
ORF038341 1.0 ug DNA
EUR 607.2

Human AIDA knockout cell line
ABC-KH0431 1 vial Ask for price
Description: Human AIDA knockout cell line is HEK293/HeLa cell line, edited by CRISPR/Cas9 technology.

Lenti ORF clone of Aida (mGFP-tagged) - Mouse axin interactor, dorsalization associated (Aida)
MR204280L4 10 µg Ask for price

Human AIDA knockdown cell line
ABC-KD0431 1 vial Ask for price
Description: Human AIDA knockdown cell line is engineered by our optimized transduction of the specific shRNA with lentivirus. Knockdown levels are determined via qRT-PCR. Gentaur offers generation of stable knockdown (RNAi) cell lines expressing shRNAs targeting genes of your interest.

Human AIDA Protein Lysate 20ug
IHUAIDAPLLY20UG each
EUR 213
Description: Human AIDA Protein Lysate 20ug

AIDA Peptide - N-terminal region
MBS3240768-01mg 0.1mg
EUR 180

AIDA Peptide - N-terminal region
MBS3240768-5x01mg 5x0.1mg
EUR 730

AIDA Peptide - N-terminal region
MBS3240769-01mg 0.1mg
EUR 180

AIDA Peptide - N-terminal region
MBS3240769-5x01mg 5x0.1mg
EUR 730

Polyclonal AIDA Antibody (C-term)
APR04233G 0.1ml
EUR 580.8
Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human AIDA (C-term). This antibody is tested and proven to work in the following applications:

AIDA antibody - N-terminal region
MBS3215866-01mL 0.1mL
EUR 455

AIDA antibody - N-terminal region
MBS3215866-5x01mL 5x0.1mL
EUR 1995

AIDA antibody - N-terminal region
MBS3215867-01mL 0.1mL
EUR 455

AIDA antibody - N-terminal region
MBS3215867-5x01mL 5x0.1mL
EUR 1995

OCOA08513-20UG - AIDA Protein Lysate
OCOA08513-20UG 20ug
EUR 169

AIDA (15O14) Rabbit Monoclonal Antibody
E28M2577 100ul
EUR 295

AIDA Over-expression Lysate Product
GWB-956789 0.1 mg Ask for price

C1orf80 (AIDA) (NM_022831) Human Over-expression Lysate
E45H11793-2 each
EUR 395

C1orf80 (AIDA) (NM_022831) Human Over-expression Lysate
LS064853 100ug
EUR 628
Description: Transient overexpression lysate of axin interactor, dorsalization associated (AIDA)

Lenti ORF clone of Aida (Myc-DDK-tagged) - Mouse axin interactor, dorsalization associated (Aida)
MR204280L3 10 µg Ask for price

Lenti ORF particles, Aida (GFP-tagged) - Mouse axin interactor, dorsalization associated (Aida), 200ul, >10^7 TU/mL
MR204280L4V 200 µl Ask for price

Lenti ORF particles, AIDA (mGFP-tagged) - Human axin interactor, dorsalization associated (AIDA), 200ul, >10^7 TU/mL
RC217521L4V 200 µl Ask for price

Kylt® EAST, AIDA, paa LD 100
31714 each
EUR 1143.41

Kylt® EAST, AIDA, paa LD 25
31715 each
EUR 359.7

AIDA sgRNA CRISPR Lentivector set (Human)
K0062101 3 x 1.0 ug
EUR 406.8

Aida sgRNA CRISPR Lentivector set (Mouse)
K4069401 3 x 1.0 ug
EUR 406.8

AIDA Antibody (C-term) Blocking Peptide
MBS9222737-INQUIRE INQUIRE Ask for price

Lenti ORF particles, Aida (Myc-DDK-tagged) - Mouse axin interactor, dorsalization associated (Aida), 200ul, >10^7 TU/mL
MR204280L3V 200 µl Ask for price

Lenti ORF particles, AIDA (Myc-DDK tagged) - Human axin interactor, dorsalization associated (AIDA), 200ul, >10^7 TU/mL
RC217521L3V 200 µl Ask for price

Aida 3'UTR GFP Stable Cell Line
TU151582 1.0 ml Ask for price

AIDA 3'UTR GFP Stable Cell Line
TU050511 1.0 ml
EUR 1825.2

AIDA, CT (AIDA, C1orf80, Axin interactor, dorsalization-associated protein, Axin interaction partner and dorsalization antagonist)
MBS6009442-02mL 0.2(mL
EUR 695

AIDA, CT (AIDA, C1orf80, Axin interactor, dorsalization-associated protein, Axin interaction partner and dorsalization antagonist)
MBS6009442-5x02mL 5x0.2mL
EUR 2975

AIDA Protein Vector (Human) (pPM-C-HA)
PV001111 500 ng
EUR 394.8

AIDA Protein Vector (Mouse) (pPM-C-HA)
PV153364 500 ng
EUR 723.6

AIDA Protein Vector (Human) (pPB-C-His)
PV001109 500 ng
EUR 394.8

AIDA Protein Vector (Human) (pPB-N-His)
PV001110 500 ng
EUR 394.8

AIDA Protein Vector (Human) (pPM-C-His)
PV001112 500 ng
EUR 394.8
The protein consists of 1136 amino acid residues encoded by 3411 bases and has a molecular weight of 125.35 kDa. The optimum temperature and pH of this enzyme are 50°C and pH 4.0. In addition, this examine confirmed that the unknown perform area (encoding Lys261-Thr681) outlined as a linker is sort of necessary for its exercise. The obtained novel enzyme BcFucA will contribute to the efficient degradation of fucoidan and future industrial functions.