High-throughput sequencing techniques were used to analyze the diversity of respiratory microbiota in health and disease. Despite the extensive data available on the respiratory tract bacterial microbiota, the remnants of the less studied fungal components. This is partly due to the technical problems associated with fungal metagenomics analysis. In this study, we compared two protocols of DNA extraction and amplification of two targets for the combined fungal bacteria and fungi that are targeted amplicon sequencing analysis of sputum microbiota.Six breathing, selected randomly from a sample routine Mondor Hospital (Créteil, France) and treated anonymously, were tested after a routine culture of bacteria and fungi.
Two of spiked Fumigati Aspergillus and Aspergillus Nigri (105 conidia / ml). After the mechanical lysis, DNA was extracted using the automatic extraction of QIAsymphony® (AQE) or users PowerSoil® MoBio extraction (MPE). DNA yield and purity compared. DNA extracted from sputum spiked into the target (i) real-time PCR for detection of Aspergillus DNA and (ii) a combination of primary targeting barcode metagenomic analysis for ITS1 and ITS2 fungi, and bacteria V1-V2 and V3-V4 region of the 16S. amplicon libraries were prepared using reagents MiSeq V3 kit on the Illumina platform. Data were analyzed using software PyroMIC © and Shaman, and compared with culture results.AQE extraction provides a higher yield of the (ratio AQE / MPE DNA = 4.5 [1.3 to 11]) DNA in a shorter period of time.
Results of Aspergillus DNA is detected by qPCR were similar to sputum spiked regardless of the extraction protocol. extraction quite an impact on biodiversity or the relative abundance of bacterial communities using targeted amplicon sequencing (2/43 taxa impact). For fungi, the relative abundance of major taxa 4/11 affected and AQE results were closer to the culture results. V1-V2 or V3-V4 and ITS1 or ITS2 target is judged diversity major taxa of bacteria and fungi, but ITS2 and V3-V4 detected smaller taxa.Our results show the importance of joint extraction of DNA for bacteria and fungi targeted metagenomics respiratory samples. Extraction protocol may affect the results of the DNA and the relative abundance of a few bacteria, but more fungal taxa. For the analysis of fungi, ITS2 allow detection of a large number of taxa small compared with ITS1.
ACTG1 human mutation associated with high frequency hearing loss, and patients with mutations in this gene are good candidates for the electric acoustic stimulation. To better understand the genetic etiology of hearing loss cases, massively parallel DNA sequencing performed on a 7048 Japanese probands related hearing loss. Among the 1,336 patients with autosomal dominant hearing loss, we identified 15 probands (1.1%) with 13 ACTG1 potentially pathogenic variant. Six new variant and seven were reported previously.
We collect and analyze detailed clinical features of these patients. The average level of development of pure-tone hearing worsening of the average for the four frequency is 1.7 dB / year of age 0-50 years, and all individuals over 60 years old has a severe hearing loss. To better understand the mechanisms underlying causes of disease, the intracellular localization of the wild-type and mutant gamma-actins checked using NIH / 3T3 fibroblast cell line
TMB Substrate, for Western Blot |
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F129-100 | Cygnus Technologies | 100 ml | EUR 249 |
Description: TMB Substrate, for Western Blot by Cygnus Technologies is available in Europe via Gentaur. |
Hypersensitive Western Blot Chemiluminescent Substrate |
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CR0001-10 | BosterBio | 20mL (sufficient reagents for 4000 cm2 of membrane) | EUR 167 |
Hypersensitive Western Blot Chemiluminescent Substrate |
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CR0001-25 | BosterBio | 50mL (sufficient reagents for 10000 cm2 of membrane) | EUR 314 |
Hypersensitive Western Blot Chemiluminescent Substrate |
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CR0001-5 | BosterBio | 10mL (sufficient reagents for 2000 cm2 of membrane) | EUR 116 |
AgileBlotter? Automated Western Blot Processor |
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ACT-ABH-40 | ACTGene | EUR 16939 |
Interferon alpha Western Blot Antibody |
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20-abx137014 | Abbexa |
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Chemiluminescent Western Blot Kit (OKRA00001) |
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OKRA00001 | Aviva Systems Biology | 1 kit | EUR 570 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Chemiluminescent Western Blot Kit (OKRA00002) |
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OKRA00002 | Aviva Systems Biology | 1 kit | EUR 570 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Chemiluminescent Western Blot Kit (OKRA00003) |
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OKRA00003 | Aviva Systems Biology | 1 kit | EUR 570 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Chemiluminescent Western Blot Kit (OKRA00004) |
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OKRA00004 | Aviva Systems Biology | 1 kit | EUR 570 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Colorimetric Western Blot Kit (OKRA00014) |
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OKRA00014 | Aviva Systems Biology | 1 kit | EUR 609 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Colorimetric Western Blot Kit (OKRA00015) |
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OKRA00015 | Aviva Systems Biology | 1 each | EUR 609 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Colorimetric Western Blot Kit (OKRA00016) |
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OKRA00016 | Aviva Systems Biology | 1 kit | EUR 609 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
Colorimetric Western Blot Kit (OKRA00017) |
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OKRA00017 | Aviva Systems Biology | 1 kit | EUR 609 |
Description: Description of target: ;Species reactivity: ;Application: ;Assay info: ;Sensitivity: |
PDCD4 Western Blot kit (AWBK09045) |
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AWBK09045 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ETS1 Western Blot kit (AWBK100604) |
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AWBK100604 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ERG Western Blot kit (AWBK100625) |
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AWBK100625 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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GABRP Western Blot kit (AWBK13034) |
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AWBK13034 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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HTR1A Western Blot kit (AWBK13041) |
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AWBK13041 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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RAB14 Western Blot kit (AWBK13107) |
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AWBK13107 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TNRC4 Western Blot kit (AWBK31525) |
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AWBK31525 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ESRRG Western Blot kit (AWBK31655) |
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AWBK31655 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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GLI2 Western Blot kit (AWBK31885) |
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AWBK31885 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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PAX4 Western Blot kit (AWBK32064) |
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AWBK32064 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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AHR Western Blot kit (AWBK32243) |
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AWBK32243 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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GLI1 Western Blot kit (AWBK32368) |
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AWBK32368 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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PAX7 Western Blot kit (AWBK32393) |
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AWBK32393 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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LHX6 Western Blot kit (AWBK32553) |
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AWBK32553 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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HIPK2 Western Blot kit (AWBK32586) |
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AWBK32586 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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PAX7 Western Blot kit (AWBK32742) |
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AWBK32742 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ZFP67 Western Blot kit (AWBK32749) |
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AWBK32749 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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OLIG2 Western Blot kit (AWBK32753) |
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AWBK32753 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ENG Western Blot kit (AWBK33068) |
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AWBK33068 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SOX5 Western Blot kit (AWBK33323) |
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AWBK33323 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SOX10 Western Blot kit (AWBK33326) |
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AWBK33326 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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RPS16 Western Blot kit (AWBK33535) |
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AWBK33535 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ATP7A Western Blot kit (AWBK33797) |
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AWBK33797 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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A1BG Western Blot kit (AWBK33810) |
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AWBK33810 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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NPM1 Western Blot kit (AWBK34094) |
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AWBK34094 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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LIG4 Western Blot kit (AWBK34122) |
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AWBK34122 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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RNF2 Western Blot kit (AWBK34290) |
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AWBK34290 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ZFP36 Western Blot kit (AWBK34385) |
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AWBK34385 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ARIH2 Western Blot kit (AWBK34418) |
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AWBK34418 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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KEAP1 Western Blot kit (AWBK34728) |
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AWBK34728 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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CLIC1 Western Blot kit (AWBK35045) |
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AWBK35045 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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KCNN2 Western Blot kit (AWBK35094) |
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AWBK35094 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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CLCN3 Western Blot kit (AWBK35504) |
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AWBK35504 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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P2RX7 Western Blot kit (AWBK35518) |
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AWBK35518 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TRPM3 Western Blot kit (AWBK35609) |
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AWBK35609 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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PITX2 Western Blot kit (AWBK35634) |
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AWBK35634 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ILF2 Western Blot kit (AWBK35731) |
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AWBK35731 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ONECUT2 Western Blot kit (AWBK35750) |
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AWBK35750 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ZFP42 Western Blot kit (AWBK36244) |
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AWBK36244 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ANXA1 Western Blot kit (AWBK36568) |
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AWBK36568 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SPP1 Western Blot kit (AWBK36677) |
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AWBK36677 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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RUNX2 Western Blot kit (AWBK36678) |
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AWBK36678 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ZBP1 Western Blot kit (AWBK36711) |
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AWBK36711 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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Bapx1 Western Blot kit (AWBK36826) |
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AWBK36826 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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G3BP Western Blot kit (AWBK37713) |
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AWBK37713 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TWIST1 Western Blot kit (AWBK37997) |
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AWBK37997 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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ID4 Western Blot kit (AWBK38058) |
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AWBK38058 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SATB1 Western Blot kit (AWBK38208) |
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AWBK38208 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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EED Western Blot kit (AWBK38384) |
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AWBK38384 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TARDBP Western Blot kit (AWBK38941) |
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AWBK38941 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TARDBP Western Blot kit (AWBK38942) |
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AWBK38942 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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BRD7 Western Blot kit (AWBK39018) |
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AWBK39018 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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JMJD3 Western Blot kit (AWBK40102) |
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AWBK40102 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SNRP70 Western Blot kit (AWBK40439) |
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AWBK40439 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SSB Western Blot kit (AWBK40462) |
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AWBK40462 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
STAU1 Western Blot kit (AWBK40537) |
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AWBK40537 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
PCBP1 Western Blot kit (AWBK40630) |
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AWBK40630 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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HNRPLL Western Blot kit (AWBK41102) |
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AWBK41102 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
MSI2 Western Blot kit (AWBK41112) |
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AWBK41112 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
HNRPA3 Western Blot kit (AWBK41195) |
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AWBK41195 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
WNT2B Western Blot kit (AWBK41254) |
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AWBK41254 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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GNB1L Western Blot kit (AWBK41328) |
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AWBK41328 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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SLC12A1 Western Blot kit (AWBK41388) |
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AWBK41388 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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TPM1 Western Blot kit (AWBK41392) |
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AWBK41392 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
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CEACAM6 Western Blot kit (AWBK41504) |
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AWBK41504 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
SLC22A1 Western Blot kit (AWBK41516) |
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AWBK41516 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
STK11 Western Blot kit (AWBK41743) |
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AWBK41743 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
OTC Western Blot kit (AWBK41766) |
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AWBK41766 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
TSHR Western Blot kit (AWBK41870) |
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AWBK41870 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
RHOBTB1 Western Blot kit (AWBK41883) |
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AWBK41883 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
KRT13 Western Blot kit (AWBK42031) |
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AWBK42031 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
IDH3A Western Blot kit (AWBK42237) |
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AWBK42237 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
SLC1A5 Western Blot kit (AWBK42247) |
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AWBK42247 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
PBEF1 Western Blot kit (AWBK42255) |
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AWBK42255 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
FST Western Blot kit (AWBK42287) |
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AWBK42287 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
SLC22A7 Western Blot kit (AWBK42306) |
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AWBK42306 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
ASPN Western Blot kit (AWBK42487) |
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AWBK42487 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
RSAD2 Western Blot kit (AWBK42696) |
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AWBK42696 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
PANX2 Western Blot kit (AWBK42778) |
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AWBK42778 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
PANX1 Western Blot kit (AWBK42783) |
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AWBK42783 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
TGFBI Western Blot kit (AWBK44268) |
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AWBK44268 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|
PTPN2 Western Blot kit (AWBK45362) |
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AWBK45362 | Aviva Systems Biology | 10 reactions | EUR 647 |
Description:
|