western blot sandwich, A capillary driven microfluidic chip for SERS based hCG detection

A capillary driven microfluidic chip for SERS based hCG detection

On this research, a capillary pushed microfluidic chip-based immunoassay was developed for the willpower of Human Chorionic Gonadotropin (hCG) protein, which is prohibited by the World Anti-Doping Company (WADA). Right here, we used antibody modified magnetic metallic natural framework nanoparticles (MMOFs) as a seize prob in urine pattern.
MMOF captured hCG was transferred in a capillary pushed microfluidic chip consisting of 4 chambers, and the interplay of MMOF with gold nanorods labelled with 5,5′-Dithiobis-(2-nitrobenzoic acid) (DTNB) as a Raman label was carried out within the capillary pushed microfluidic chip. The motion of MMOF by means of first chamber to the final chamber was achieved with a easy magnet.
Within the final chamber of capillary pushed microfluidic chip, SERS indicators of DTNB molecules from the sandwich advanced have been recorded utilizing a Raman spectrophotometer. The selectivity of the developed methodology was demonstrated by making use of the identical process for the detection of Human Luteinizing Hormone (hLH), Human Chorionic Gonadotropin Hormone (hGH) and Immunoglobulin G (IgG) protein.
The regression coefficient and restrict of detection obtained from the usual addition methodology have been discovered as 0,9985 and 0,61 IU/L, respectively. Moreover, the traditional ELISA methodology confirmed that the outcomes obtained by the offered methodology have been acceptable with the similarity of 97.9% when it comes to common restoration worth, for the detection of hCG in urine samples. The evaluation system developed for goal proteins might be an alternate approach comparable to Western Blot utilized in routine evaluation that’s costly and time consuming.

A easy unlabeled human chorionic gonadotropin biosensor based mostly on a peptide aptamer

As an important biochemical indicator within the fields of being pregnant and oncology, human chorionic gonadotropin (HCG) may be evaluated utilizing colloidal gold immunochromatographic paper and quantified utilizing a biochemical analyzer based mostly on the precept of the antibody sandwich methodology. In view of the inaccuracy of the previous and the complication of the latter, this research constructed an correct, delicate and easy unlabeled biosensor based mostly on peptide aptamer CGGGPPLRINRHILTR for HCG detection.
Molecular Working Surroundings (MOE) was used to simulate the aptamer and protein docking, and western blot (WB) was used to confirm the binding impact and ratio. The peptide aptamer was characterised and was then decreased with tris-(2-carboxyethyl)-phosphine hydrochloride (TCEP). After electrochemical deposition of chloroauric acid on the screen-printed electrode (SPE), the aptamer was self-assembled on the electrode floor below optimum situations.
The energetic website of the electrode floor was blocked with 6-mercapto-1-hexanol (MCH) and BSA. The electrochemical impedance spectrum (EIS) was used to quantify HCG within the matrix. Displaying a very good linear relationship within the vary of 5-1500 mIU mL-1, with a detection restrict of 1 mIU mL-1, the biosensor remained secure at room temperature for 14 days. Due to its small dimension, stability, sensitivity and accuracy, this biosensor has nice potential to grow to be a transportable diagnostic machine for HCG.

Augmentation of RBP4/STRA6 signaling results in insulin resistance and irritation and the believable therapeutic position of vildagliptin and metformin

A task of Retinol Binding Protein-4 (RBP4) in insulin resistance is extensively studied. Nonetheless, there’s paucity of data on its receptor viz., Stimulated by Retinoic Acid-6 (STRA6) with insulin resistance. To deal with this, we investigated the regulation of RBP4/STRA6 expression in 3T3-L1 adipocytes uncovered to glucolipotoxicity (GLT) and in visceral adipose tissue (VAT) from excessive fats eating regimen (HFD) fed insulin-resistant rats.
3T3-L1 adipocytes have been subjected to GLT and different experimental maneuvers with and with out vildagliptin or metformin. Actual-time PCR and westernblot experiments have been carried out to research RBP4, STRA6, PPARγ gene and protein expression.
Adipored staining and glucose uptake assay have been carried out to guage lipid and glucose metabolism. Oral glucose tolerance check (OGTT) and Insulin Tolerance Take a look at (ITT) have been carried out to find out the extent of insulin resistance in HFD fed male Wistar rats. Whole serum RBP4 was measured by quantitative sandwich enzyme-linked immunosorbent assay equipment.
Adipocytes below GLT exhibited considerably elevated RBP4/STRA6 expressions and decreased insulin sensitivity/glucose uptake. Vildagliptin and metformin not solely restored the above but in addition decreased the expression of IL-6, NFκB, SOCS-Three together with lipid accumulation.
Moreover, HFD fed rats exhibited considerably elevated serum ranges of RBP4 together with VAT expression of RBP4, STRA6, PPARγ, IL-6. These molecules have been considerably altered by the vildagliptin/ metformin remedy. We conclude that RBP4/STRA6 pathway is primarily concerned in mediating irritation and insulin resistance in adipocytes and visceral adipose tissues below glucolipotoxicity and in insulin resistant rats.

Structural and biochemical characterization of a novel thermophilic Coh01147 protease

Proteases play an important position in dwelling organisms and characterize one of many largest teams of business enzymes. The intention of this work was recombinant manufacturing and characterization of a newly recognized thermostable protease 1147 from thermophilum indigenous Cohnella sp. A01.
Phylogenetic tree evaluation confirmed that protease 1147 is intently associated to the cysteine proteases from DJ-1/ThiJ/PfpI superfamily, with the conserved catalytic tetrad. Structural prediction utilizing MODELLER 9v7 indicated that protease 1147 has an general α/β sandwich tertiary construction.
western blot sandwich, A capillary driven microfluidic chip for SERS based hCG detection
The gene of protease 1147 was cloned and expressed in Escherichia coli (E. coli) BL21. The recombinant protease 1147 appeared as a homogenous band of 18 kDa in SDS-PAGE, which was verified by western blot and zymography. The recombinant protein was purified with a yield of roughly 88% in a single step utilizing Ni-NTA affinity chromatography.
Moreover, a fast one-step thermal shock process was efficiently applied to purify the protein with a yield of 73%. Utilizing casein because the substrate, Km, and kcat, kcat/Km values of 13.72 mM, 3.143 × 10-3 (s-1), and 0.381 (M-1 S-1) have been obtained, respectively.
The utmost protease exercise was detected at pH = 7 and 60°C with the inactivation fee fixed (kin) of two.10 × 10-3 (m-1), and half-life (t1/2) of 330.07 min. Protease 1147 exhibited wonderful stability to natural solvent, metallic ions, and 1% SDS. The protease exercise was considerably enhanced by Tween 20 and Tween 80 and suppressed by cysteine protease particular inhibitors.

Hypersensitive Western Blot Chemiluminescent Substrate

CR0001-10 20mL (sufficient reagents for 4000 cm2 of membrane)
EUR 167

Hypersensitive Western Blot Chemiluminescent Substrate

CR0001-25 50mL (sufficient reagents for 10000 cm2 of membrane)
EUR 314

Hypersensitive Western Blot Chemiluminescent Substrate

CR0001-5 10mL (sufficient reagents for 2000 cm2 of membrane)
EUR 116

AgileBlotter? Automated Western Blot Processor

ACT-ABH-40
EUR 16939

TMB Substrate, for Western Blot

F100029 1000 ml
EUR 827
  • Product category: Buffer solution
Description: TMB Substrate, for Western Blot by Cygnus Technologies is available in Europe via Gentaur.

TMB Substrate, for Western Blot

F129-100 100 ml
EUR 249
  • Product category: Buffer solution
Description: TMB Substrate, for Western Blot by Cygnus Technologies is available in Europe via Gentaur.

PDCD4 Western Blot kit (AWBK09045)

AWBK09045 10 reactions
EUR 647
Description:
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ETS1 Western Blot kit (AWBK100604)

AWBK100604 10 reactions
EUR 647
Description:
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ERG Western Blot kit (AWBK100625)

AWBK100625 10 reactions
EUR 647
Description:
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GABRP Western Blot kit (AWBK13034)

AWBK13034 10 reactions
EUR 647
Description:
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HTR1A Western Blot kit (AWBK13041)

AWBK13041 10 reactions
EUR 647
Description:
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RAB14 Western Blot kit (AWBK13107)

AWBK13107 10 reactions
EUR 647
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TNRC4 Western Blot kit (AWBK31525)

AWBK31525 10 reactions
EUR 647
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ESRRG Western Blot kit (AWBK31655)

AWBK31655 10 reactions
EUR 647
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GLI2 Western Blot kit (AWBK31885)

AWBK31885 10 reactions
EUR 647
Description:
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PAX4 Western Blot kit (AWBK32064)

AWBK32064 10 reactions
EUR 647
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AHR Western Blot kit (AWBK32243)

AWBK32243 10 reactions
EUR 647
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GLI1 Western Blot kit (AWBK32368)

AWBK32368 10 reactions
EUR 647
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PAX7 Western Blot kit (AWBK32393)

AWBK32393 10 reactions
EUR 647
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LHX6 Western Blot kit (AWBK32553)

AWBK32553 10 reactions
EUR 647
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HIPK2 Western Blot kit (AWBK32586)

AWBK32586 10 reactions
EUR 647
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PAX7 Western Blot kit (AWBK32742)

AWBK32742 10 reactions
EUR 647
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ZFP67 Western Blot kit (AWBK32749)

AWBK32749 10 reactions
EUR 647
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OLIG2 Western Blot kit (AWBK32753)

AWBK32753 10 reactions
EUR 647
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ENG Western Blot kit (AWBK33068)

AWBK33068 10 reactions
EUR 647
Description:
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SOX5 Western Blot kit (AWBK33323)

AWBK33323 10 reactions
EUR 647
Description:
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SOX10 Western Blot kit (AWBK33326)

AWBK33326 10 reactions
EUR 647
Description:
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RPS16 Western Blot kit (AWBK33535)

AWBK33535 10 reactions
EUR 647
Description:
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ATP7A Western Blot kit (AWBK33797)

AWBK33797 10 reactions
EUR 647
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A1BG Western Blot kit (AWBK33810)

AWBK33810 10 reactions
EUR 647
Description:
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  • Species reactivity: Human
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NPM1 Western Blot kit (AWBK34094)

AWBK34094 10 reactions
EUR 647
Description:
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LIG4 Western Blot kit (AWBK34122)

AWBK34122 10 reactions
EUR 647
Description:
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RNF2 Western Blot kit (AWBK34290)

AWBK34290 10 reactions
EUR 647
Description:
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ZFP36 Western Blot kit (AWBK34385)

AWBK34385 10 reactions
EUR 647
Description:
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ARIH2 Western Blot kit (AWBK34418)

AWBK34418 10 reactions
EUR 647
Description:
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KEAP1 Western Blot kit (AWBK34728)

AWBK34728 10 reactions
EUR 647
Description:
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CLIC1 Western Blot kit (AWBK35045)

AWBK35045 10 reactions
EUR 647
Description:
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KCNN2 Western Blot kit (AWBK35094)

AWBK35094 10 reactions
EUR 647
Description:
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CLCN3 Western Blot kit (AWBK35504)

AWBK35504 10 reactions
EUR 647
Description:
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P2RX7 Western Blot kit (AWBK35518)

AWBK35518 10 reactions
EUR 647
Description:
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TRPM3 Western Blot kit (AWBK35609)

AWBK35609 10 reactions
EUR 647
Description:
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PITX2 Western Blot kit (AWBK35634)

AWBK35634 10 reactions
EUR 647
Description:
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ILF2 Western Blot kit (AWBK35731)

AWBK35731 10 reactions
EUR 647
Description:
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ONECUT2 Western Blot kit (AWBK35750)

AWBK35750 10 reactions
EUR 647
Description:
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ZFP42 Western Blot kit (AWBK36244)

AWBK36244 10 reactions
EUR 647
Description:
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ANXA1 Western Blot kit (AWBK36568)

AWBK36568 10 reactions
EUR 647
Description:
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SPP1 Western Blot kit (AWBK36677)

AWBK36677 10 reactions
EUR 647
Description:
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RUNX2 Western Blot kit (AWBK36678)

AWBK36678 10 reactions
EUR 647
Description:
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ZBP1 Western Blot kit (AWBK36711)

AWBK36711 10 reactions
EUR 647
Description:
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Bapx1 Western Blot kit (AWBK36826)

AWBK36826 10 reactions
EUR 647
Description:
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G3BP Western Blot kit (AWBK37713)

AWBK37713 10 reactions
EUR 647
Description:
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TWIST1 Western Blot kit (AWBK37997)

AWBK37997 10 reactions
EUR 647
Description:
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ID4 Western Blot kit (AWBK38058)

AWBK38058 10 reactions
EUR 647
Description:
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SATB1 Western Blot kit (AWBK38208)

AWBK38208 10 reactions
EUR 647
Description:
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EED Western Blot kit (AWBK38384)

AWBK38384 10 reactions
EUR 647
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TARDBP Western Blot kit (AWBK38941)

AWBK38941 10 reactions
EUR 647
Description:
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TARDBP Western Blot kit (AWBK38942)

AWBK38942 10 reactions
EUR 647
Description:
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BRD7 Western Blot kit (AWBK39018)

AWBK39018 10 reactions
EUR 647
Description:
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JMJD3 Western Blot kit (AWBK40102)

AWBK40102 10 reactions
EUR 647
Description:
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SNRP70 Western Blot kit (AWBK40439)

AWBK40439 10 reactions
EUR 647
Description:
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SSB Western Blot kit (AWBK40462)

AWBK40462 10 reactions
EUR 647
Description:
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STAU1 Western Blot kit (AWBK40537)

AWBK40537 10 reactions
EUR 647
Description:
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PCBP1 Western Blot kit (AWBK40630)

AWBK40630 10 reactions
EUR 647
Description:
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HNRPLL Western Blot kit (AWBK41102)

AWBK41102 10 reactions
EUR 647
Description:
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MSI2 Western Blot kit (AWBK41112)

AWBK41112 10 reactions
EUR 647
Description:
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HNRPA3 Western Blot kit (AWBK41195)

AWBK41195 10 reactions
EUR 647
Description:
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WNT2B Western Blot kit (AWBK41254)

AWBK41254 10 reactions
EUR 647
Description:
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GNB1L Western Blot kit (AWBK41328)

AWBK41328 10 reactions
EUR 647
Description:
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SLC12A1 Western Blot kit (AWBK41388)

AWBK41388 10 reactions
EUR 647
Description:
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TPM1 Western Blot kit (AWBK41392)

AWBK41392 10 reactions
EUR 647
Description:
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CEACAM6 Western Blot kit (AWBK41504)

AWBK41504 10 reactions
EUR 647
Description:
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SLC22A1 Western Blot kit (AWBK41516)

AWBK41516 10 reactions
EUR 647
Description:
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STK11 Western Blot kit (AWBK41743)

AWBK41743 10 reactions
EUR 647
Description:
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OTC Western Blot kit (AWBK41766)

AWBK41766 10 reactions
EUR 647
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TSHR Western Blot kit (AWBK41870)

AWBK41870 10 reactions
EUR 647
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RHOBTB1 Western Blot kit (AWBK41883)

AWBK41883 10 reactions
EUR 647
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KRT13 Western Blot kit (AWBK42031)

AWBK42031 10 reactions
EUR 647
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IDH3A Western Blot kit (AWBK42237)

AWBK42237 10 reactions
EUR 647
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SLC1A5 Western Blot kit (AWBK42247)

AWBK42247 10 reactions
EUR 647
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PBEF1 Western Blot kit (AWBK42255)

AWBK42255 10 reactions
EUR 647
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FST Western Blot kit (AWBK42287)

AWBK42287 10 reactions
EUR 647
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SLC22A7 Western Blot kit (AWBK42306)

AWBK42306 10 reactions
EUR 647
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ASPN Western Blot kit (AWBK42487)

AWBK42487 10 reactions
EUR 647
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RSAD2 Western Blot kit (AWBK42696)

AWBK42696 10 reactions
EUR 647
Description:
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PANX2 Western Blot kit (AWBK42778)

AWBK42778 10 reactions
EUR 647
Description:
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PANX1 Western Blot kit (AWBK42783)

AWBK42783 10 reactions
EUR 647
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TGFBI Western Blot kit (AWBK44268)

AWBK44268 10 reactions
EUR 647
Description:
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PTPN2 Western Blot kit (AWBK45362)

AWBK45362 10 reactions
EUR 647
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RORA Western Blot kit (AWBK45608)

AWBK45608 10 reactions
EUR 647
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Chemiluminescent Western Blot Kit (OKRA00001)

OKRA00001 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00002)

OKRA00002 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00003)

OKRA00003 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Chemiluminescent Western Blot Kit (OKRA00004)

OKRA00004 1 kit
EUR 570
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00014)

OKRA00014 1 kit
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00015)

OKRA00015 1 each
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:

Colorimetric Western Blot Kit (OKRA00016)

OKRA00016 1 kit
EUR 609
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity:
Docking outcomes and molecular dynamics (MD) simulation revealed that Tween 20 interacted with protease 1147 through hydrogen bonds and made the construction extra secure. CD and fluorescence spectra indicated structural adjustments happening at 100°C, very primary and acidic pH, and within the presence of Tween 20. These properties make this newly characterised protease a possible candidate for varied biotechnological functions.

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