Right here, we explored whether or not a minicircle DNA (MC) vector containing human angiopoietin 1 (MC-ANGPT1) can present a more practical platform for gene-enhanced MSC remedy of ALI/ARDS. At 24 h after transfection, nuclear-targeted electroporation utilizing an MC-ANGPT1 vector resulted in a 3.7-fold better enhance in human ANGPT1 protein in MSC conditioned media in comparison with the usage of a plasmid ANGPT1 vector. Within the lipopolysaccharide (LPS)-induced ALI mannequin, administration of pANGPT1 transfected MSCs considerably decreased bronchoalveolar lavage (BAL) neutrophil counts by 57%, whereas MC-ANGPT1 transfected MSCs decreased it by 71% (p < 0.001) by Holm-Sidak’s a number of comparability take a look at.
Furthermore, in comparison with pANGPT1, the MC-ANGPT1 transfected MSCs considerably decreased pulmonary irritation, as noticed in decreased ranges of proinflammatory cytokines, akin to tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and macrophage inflammatory protein-2 (MIP-2). pANGPT1-transfected MSCs considerably decreased BAL albumin ranges by 71%, whereas MC-ANGPT1-transfected MSCs decreased it by 85%. Total, utilizing a minicircle vector, we demonstrated an environment friendly and sustained expression of the ANGPT1 transgene in MSCs and enhanced the therapeutic impact on the ALI mannequin in comparison with plasmid.
Plasmid Vectors for in Vivo Choice-Free Use with the Probiotic E. coli Nissle 1917
Escherichia coli Nissle 1917 (EcN) is a probiotic bacterium, generally employed to deal with sure gastrointestinal issues. It’s quick rising as an essential goal for the event of therapeutic engineered micro organism, benefiting from the wealth of information of E. coli biology and ease of manipulation. Bacterial artificial biology initiatives generally make the most of engineered plasmid vectors, that are easy to engineer and might reliably obtain excessive ranges of protein expression. Nevertheless, plasmids usually require antibiotics for upkeep, and the administration of an antibiotic is commonly incompatible with in vivo experimentation or remedy.
EcN natively accommodates plasmids pMUT1 and pMUT2, which don’t have any recognized operate however are secure inside the micro organism. Right here, we describe the event of the pMUT plasmids into a strong platform for engineering EcN for in vivo experimentation, alongside a CRISPR-Cas9 system to take away the native plasmids. We systematically engineered each pMUT plasmids to include choice markers, fluorescent markers, temperature delicate expression, and curli secretion programs to export a customizable purposeful materials into the extracellular area.
We then reveal that the engineered plasmids had been maintained in micro organism because the engineered micro organism go by the mouse GI tract with out choice, and that the secretion system stays purposeful, exporting functionalized curli proteins into the intestine. Our plasmid system presents a platform for the fast improvement of therapeutic EcN micro organism.
Cloning in Plasmid Vectors: Blunt-Finish Cloning
This protocol describes procedures for cloning blunt-ended DNA fragments into linearized plasmid vectors. To acquire the utmost variety of “appropriate” ligation merchandise when cloning blunt-ended goal fragments, the 2 elements of DNA within the ligation response have to be current at an acceptable ratio. These outcomes assist the potential advantages of MC-ANGPT1 gene enhancement of MSC remedy to deal with ARDS.
If the molar ratio of plasmid vector to focus on DNA is just too excessive, then the ligation response might generate an undesirable variety of round empty plasmids, each monomeric and polymeric; if too low, the ligation response might generate an extra of linear and round homopolymers and heteropolymers of various sizes, orientations, and compositions. Because of this, the orientation of the overseas DNA and the variety of inserts in every recombinant clone should at all times be validated by restriction endonuclease mapping or another means.
Plasmid copy quantity variation of a modular vector set in Shewanella oneidensis MR-1
To advance artificial biology approaches that make the most of S. oneidensis as host for biotechnology functions, now we have investigated the variation in plasmid copy variety of a modular vector set ensuing from distinct origins of replication beneath totally different circumstances. The replicons yielded a ≈9X-fold vary for plasmid copy quantity variation in S. oneidensis (whereas the identical origins yielded a ≈3X-fold vary in Escherichia coli). This offers a sizeable vary to regulate gene expression ranges in S. oneidensis for artificial biology functions. As well as, plasmid harboring the pBBR1 origin resulted in secure copy numbers in S. oneidensis beneath totally different circumstances (mid-logarithmic, stationary, multi-plasmid).
NFKB3 Antibody |
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MBS859366-01mLAF610 | MyBiosource | 0.1mL(AF610) | EUR 565 |
NFKB3 Antibody |
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MBS859366-01mLAF635 | MyBiosource | 0.1mL(AF635) | EUR 565 |
NFKB3 Antibody |
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MBS855382-01mg | MyBiosource | 0.1mg | EUR 345 |
NFKB3 Antibody |
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MBS855382-01mLAF405L | MyBiosource | 0.1mL(AF405L) | EUR 565 |
NFKB3 Antibody |
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MBS855382-01mLAF405S | MyBiosource | 0.1mL(AF405S) | EUR 565 |
NFKB3 Antibody |
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MBS855382-01mLAF610 | MyBiosource | 0.1mL(AF610) | EUR 565 |
NFKB3 Antibody |
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MBS855382-01mLAF635 | MyBiosource | 0.1mL(AF635) | EUR 565 |
Transcription Factor P65 (NFKB3) Antibody |
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abx037885-100ug | Abbexa | 100 ug | EUR 469.2 |
Transcription Factor P65 (NFKB3) Antibody |
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abx038310-100ug | Abbexa | 100 ug | EUR 469.2 |
Transcription Factor P65 (NFKB3) Antibody |
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abx037885-100g | Abbexa | 100 µg | EUR 337.5 |
Transcription Factor P65 (NFKB3) Antibody |
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abx038310-1mg | Abbexa | 1 mg | EUR 337.5 |
Transcription Factor P65 (NFKB3) Polyclonal Antibody |
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CAU27518-100ul | Biomatik Corporation | 100ul | EUR 217.9 |
Transcription Factor P65 (NFKB3) Polyclonal Antibody |
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CAU27518-200ul | Biomatik Corporation | 200ul | EUR 272.2 |
Transcription Factor P65 (NFKB3) Polyclonal Antibody |
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CAU27519-100ul | Biomatik Corporation | 100ul | EUR 212.2 |
Transcription Factor P65 (NFKB3) Polyclonal Antibody |
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CAU27519-200ul | Biomatik Corporation | 200ul | EUR 265.4 |
Polyclonal Antibody to Transcription Factor P65 (NFKB3) |
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PAA616Hu01 | Cloud-Clone | 100ul | EUR 221 |
Polyclonal Antibody to Transcription Factor P65 (NFKB3) |
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PAA616Mu01 | Cloud-Clone | 100ul | EUR 227 |
Polyclonal Antibody to Transcription Factor P65 (NFKB3) |
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PAA616Ra01 | Cloud-Clone | 100ul | EUR 239 |
anti- NFKB1 antibody |
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FNab10143 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against NFKB1 |
anti- NFKB1 antibody |
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FNab05702 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against NFKB1 |
anti- NFKB1 antibody |
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FNab05703 | FN Test | 100µg | EUR 702 |
Description: Antibody raised against NFKB1 |
anti- NFKB2 antibody |
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FNab05705 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against NFKB2 |
anti- NFKB2 antibody |
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FNab05706 | FN Test | 100µg | EUR 658.5 |
Description: Antibody raised against NFKB2 |
Anti- p52 (NFkB2) Antibody |
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GWB-AB58EB | GenWay Biotech | 0.1 ml | Ask for price |
Anti- p50 (NFKB1) Antibody |
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GWB-747B10 | GenWay Biotech | 0.1 ml | Ask for price |
Anti-NFKB3 Antibody |
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39294 | SAB | 100ul | EUR 479 |
Anti-NFKB3 Antibody |
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Y058181 | ABM | 100 µl | EUR 145 |
Anti-NFKB p50 (NFKB1) Antibody |
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GWB-CC6745 | GenWay Biotech | 100 ul | Ask for price |
Anti-NFkB p105/p50/NFKB1 Antibody |
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PB9149 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-NFkB p100/p52/NFKB2 Antibody |
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PB9150 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-NFkB p100/p52/NFKB2 Antibody |
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PA1946 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-NFkB p100/p52/NFKB2 Antibody |
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PA1946-1 | BosterBio | 100ug/vial | EUR 352.8 |
Anti-NFkB p105/p50/NFKB1 Antibody |
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PA2111 | BosterBio | 100ug/vial | EUR 352.8 |
Anti-NFkB p105/p50/NFKB1 Antibody |
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PA2111-1 | BosterBio | 100ug/vial | EUR 352.8 |
Recombinant NFkB3 (RELA/p65) protein |
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MBS389143-002mg | MyBiosource | 0.02mg | EUR 610 |
Recombinant NFkB3 (RELA/p65) protein |
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MBS389143-1mg | MyBiosource | 1mg | EUR 4505 |
Recombinant NFkB3 (RELA/p65) protein |
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MBS389143-5x1mg | MyBiosource | 5x1mg | EUR 20075 |
Anti- cRel (NFkB) Antibody |
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GWB-393707 | GenWay Biotech | 0.1 ml | Ask for price |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU55569-100ug | Biomatik Corporation | 100ug | EUR 918.5 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU55569-1mg | Biomatik Corporation | 1mg | EUR 3606.2 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU55569-50ug | Biomatik Corporation | 50ug | EUR 554.9 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU54244-100ug | Biomatik Corporation | 100ug | EUR 589.6 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU54244-1mg | Biomatik Corporation | 1mg | EUR 2610.4 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU54244-50ug | Biomatik Corporation | 50ug | EUR 473 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU44376-100ug | Biomatik Corporation | 100ug | EUR 393.8 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU44376-1mg | Biomatik Corporation | 1mg | EUR 1747.2 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPU44376-50ug | Biomatik Corporation | 50ug | EUR 316.8 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPA616Fi01 | Cloud-Clone | 10ug | EUR 231 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPA616Hu01 | Cloud-Clone | 10ug | EUR 191 |
Recombinant Transcription Factor P65 (NFKB3) |
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RPA616Mu01 | Cloud-Clone | 10ug | EUR 128 |
anti- NFKB1,p105,p50-Specific antibody |
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FNab05704 | FN Test | 100µg | EUR 702 |
Description: Antibody raised against NFKB1,p105,p50-Specific |
anti- NFKB2,p52,p100-Specific antibody |
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FNab05707 | FN Test | 100µg | EUR 658.5 |
Description: Antibody raised against NFKB2,p52,p100-Specific |
anti- NFKBIE antibody |
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FNab05708 | FN Test | 100µg | EUR 658.5 |
Description: Antibody raised against NFKBIE |
anti- NFKBIZ antibody |
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FNab05709 | FN Test | 100µg | EUR 606.3 |
Description: Antibody raised against NFKBIZ |
Anti-Phospho-N kappa-p105 (S893) NFKB1 Antibody |
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A00283S893 | BosterBio | 100ul | EUR 380 |
Description: Boster Bio Anti-Phospho-N kappa-p105 (S893) NFKB1 Antibody catalog # A00283S893. Tested in ELISA, IHC, WB applications. This antibody reacts with Human. |
Anti-Phospho-N kappa-p105 (S907) NFKB1 Antibody |
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A00283S907 | BosterBio | 100ul | EUR 380 |
Description: Boster Bio Anti-Phospho-N kappa-p105 (S907) NFKB1 Antibody catalog # A00283S907. Tested in ELISA, IP, IHC, WB applications. This antibody reacts with Human. |
Anti-Phospho-N kappa-p105 (S932) NFKB1 Antibody |
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A00283S932 | BosterBio | 100ul | EUR 380 |
Description: Boster Bio Anti-Phospho-N kappa-p105 (S932) NFKB1 Antibody catalog # A00283S932. Tested in ELISA, IHC, WB applications. This antibody reacts with Human, Mouse, Rat. |
Anti-Phospho-N kappa-p100 (S872) NFKB2 Antibody |
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A01228S872 | BosterBio | 100ul | EUR 380 |
Description: Boster Bio Anti-Phospho-N kappa-p100 (S872) NFKB2 Antibody catalog # A01228S872. Tested in ELISA, WB applications. This antibody reacts with Human, Mouse. |
Rabbit Anti-Human NFKB1 (aa 930-934) Polyclonal Antibody |
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CPB-1129RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB1 (aa 160-170) Polyclonal Antibody |
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CPB-1169RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB2 (aa 160-171) Polyclonal Antibody |
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CPB-1176RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit anti IkBa (NFKB1A) Polyclonal Antibody |
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TA354676 | Origene Technologies GmbH | 100 µg | Ask for price |
Human Transcription factor p65 protein (NFKB3) |
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1-CSB-RP039844h | Cusabio |
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Description: Recombinant Human Transcription factor p65 protein(NFKB3),partial expressed in E.coli |
Human Transcription factor p65 protein (NFKB3) |
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1-CSB-RP039874h | Cusabio |
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Description: Recombinant Human Transcription factor p65 protein(NFKB3),partial expressed in E.coli |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49304-48T | Biomatik Corporation | 48T | EUR 337.82 |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49304-5x96T | Biomatik Corporation | 5x96T | EUR 2292.35 |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49304-96T | Biomatik Corporation | 96T | EUR 482.6 |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08193-48T | Biomatik Corporation | 48T | EUR 542.78 |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08193-5x96T | Biomatik Corporation | 5x96T | EUR 3683.15 |
Rat Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08193-96T | Biomatik Corporation | 96T | EUR 775.4 |
ELISA Kit for Transcription Factor P65 (NFKB3) |
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SEA616Hu | Cloud-Clone | 96Т | EUR 630 |
ELISA Kit for Transcription Factor P65 (NFKB3) |
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SEA616Ra | Cloud-Clone | 96Т | EUR 684 |
Rabbit Anti-Human NFKB2 Polyclonal Antibody, aa.864-868 |
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CPB-930RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB2 Polyclonal Antibody, aa.868-872 |
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CPB-931RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB1 Polyclonal Antibody, aa.335-339 |
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CPB-932RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB1 Polyclonal Antibody, aa.891-895 |
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CPB-933RH | Creative Diagnostics | 100 ul | EUR 552 |
Rabbit Anti-Human NFKB1 Polyclonal Antibody, aa.905-909 |
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CPB-934RH | Creative Diagnostics | 100 ul | EUR 552 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49303-48T | Biomatik Corporation | 48T | EUR 311.15 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49303-5x96T | Biomatik Corporation | 5x96T | EUR 2111.38 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN49303-96T | Biomatik Corporation | 96T | EUR 444.5 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU11278-48T | Biomatik Corporation | 48T | EUR 600.25 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU11278-5x96T | Biomatik Corporation | 5x96T | EUR 4073.13 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU11278-96T | Biomatik Corporation | 96T | EUR 857.5 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08192-48T | Biomatik Corporation | 48T | EUR 500.5 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08192-5x96T | Biomatik Corporation | 5x96T | EUR 3396.25 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKU08192-96T | Biomatik Corporation | 96T | EUR 715 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN52389-48T | Biomatik Corporation | 48T | EUR 373.38 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN52389-5x96T | Biomatik Corporation | 5x96T | EUR 2533.65 |
Human Transcription Factor P65 (NFKB3) ELISA Kit |
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EKN52389-96T | Biomatik Corporation | 96T | EUR 533.4 |
Rabbit anti IkBa (pS32/36) (NFKB1A) Polyclonal Antibody |
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TA354674 | Origene Technologies GmbH | 100 µg | Ask for price |
Anti- NFkB p65 (Rel A) Antibody |
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GWB-34C0DE | GenWay Biotech | 0.1 ml | Ask for price |
Anti-IKBL1 NFKBIL1 Antibody |
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A05691-1 | BosterBio | 100ul | EUR 380 |
Description: Boster Bio Anti-IKBL1 NFKBIL1 Antibody catalog # A05691-1. Tested in ELISA, WB applications. This antibody reacts with Human, Mouse, Rat. |
NFKB2 Rabbit anti-Human Polyclonal (aa38-343) Antibody |
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MBS2402498-005mL | MyBiosource | 0.05mL | EUR 615 |
NFKB2 Rabbit anti-Human Polyclonal (aa38-343) Antibody |
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MBS2402498-5x005mL | MyBiosource | 5x0.05mL | EUR 2605 |
Rabbit anti IkBa (Paired S32/36) (NFKB1A) Polyclonal Antibody |
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TA354675 | Origene Technologies GmbH | 100 µg | Ask for price |
Anti-IKB beta/NFKBIB Antibody |
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PB9292 | BosterBio | 100ug/vial | EUR 400.8 |
Anti-IKB alpha/NFKBIA Antibody |
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PB9291 | BosterBio | 100ug/vial | EUR 400.8 |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx255851-96tests | Abbexa | 96 tests | EUR 801.6 |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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20-abx156040 | Abbexa |
|
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Pig Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx361108-96tests | Abbexa | 96 tests | EUR 990 |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx255851-1096tests | Abbexa | 10 × 96 tests | Ask for price |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx255851-596tests | Abbexa | 5 × 96 tests | Ask for price |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx156040-100g | Abbexa | 100 µg | EUR 6075 |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx156040-10g | Abbexa | 10 µg | EUR 675 |
Rat Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx156040-50g | Abbexa | 50 µg | EUR 3212.5 |
Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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20-abx152945 | Abbexa |
|
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Mouse Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx254289-96tests | Abbexa | 96 tests | EUR 801.6 |
Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx252815-96tests | Abbexa | 96 tests | EUR 801.6 |
Human Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx570125-96tests | Abbexa | 96 tests | EUR 801.6 |
Mouse Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx570127-96tests | Abbexa | 96 tests | EUR 801.6 |
Sheep Transcription factor p65 / NFKB3 (RELA) ELISA Kit |
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abx364265-96tests | Abbexa | 96 tests | EUR 1111.2 |
Rabbit Anti-Human NFKB1 Polyclonal Antibody, Phospho-Ser932 |
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CPB-782RH | Creative Diagnostics | 100 ul | EUR 670.8 |
Rabbit Anti-Human NFKB1 Polyclonal Antibody, Phospho-Ser927 |
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CPB-827RH | Creative Diagnostics | 100 ul | EUR 670.8 |
NFKB1 antibody |
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10R-11003 | Fitzgerald | 100 ug | EUR 344 |
Description: Mouse monoclonal NFKB1 antibody |
NFKB1 antibody |
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10R-11155 | Fitzgerald | 100 ug | EUR 344 |
Description: Mouse Monoclonal NFKB1 antibody |
NFKB2 antibody |
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10R-1605 | Fitzgerald | 100 ug | EUR 614.4 |
Description: Mouse monoclonal NFKB2 antibody |
NFKB1 antibody |
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20-PR03 | Fitzgerald | 100 ul | Ask for price |
Description: Rabbit polyclonal NFKB1 antibody |
NFKB2 Antibody |
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37182 | SAB | 100ul | EUR 319 |
NFKB2 Antibody |
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37182-100ul | SAB | 100ul | EUR 302.4 |
NFKB1 Antibody |
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35588 | SAB | 100ul | EUR 319 |
NFKB1 Antibody |
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35588-100ul | SAB | 100ul | EUR 302.4 |
NFKB2 antibody |
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38583 | SAB | 100ul | EUR 439 |
This will allow the belief of artificial circuits in S. oneidensis the place predictable, quantitative conduct is desired (in both single- or double-plasmid contexts). Klebsiella pneumoniae is a bacterial pathogen with rising charges of resistance to carbapenem antibiotics, however the inhabitants construction and genetic drivers of carbapenem-resistant Okay pneumoniae (CRKP) stay underexplored in creating nations. Carbapenem-resistant Okay pneumoniae had been not too long ago launched into Peru however have grown quickly in prevalence, enabling research of this pathogen because it expands into an unaffected setting.