UFC901024 0000234282
- Document Type:
- Certificate of Quality
- Lot Number:
- 0000234282
- Product Catalog Number:
- UFC901024
- Product Catalog Name:
- Amicon® Ultra-15 Centrifugal Filter Unit
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UFC910024 - 0000241948
- Document Type:
- Certificate of Quality
- Lot Number:
- 0000241948
- Product Catalog Number:
- UFC910024
- Product Catalog Name:
- Amicon® Ultra-15 Centrifugal Filter Unit
-
Cross-repressive interactions between Lrig3 and netrin 1 shape the architecture of the inner ear.
- The sense of balance depends on the intricate architecture of the inner ear, which contains three semicircular canals used to detect motion of the head in space. Changes in the shape of even one canal cause drastic behavioral deficits, highlighting the need to understand the cellular and molecular events that ensure perfect formation of this precise structure. During development, the canals are sculpted from pouches that grow out of a simple ball of epithelium, the otic vesicle. A key event is the fusion of two opposing epithelial walls in the center of each pouch, thereby creating a hollow canal. During the course of a gene trap mutagenesis screen to find new genes required for canal morphogenesis, we discovered that the Ig superfamily protein Lrig3 is necessary for lateral canal development. We show that this phenotype is due to ectopic expression of the axon guidance molecule netrin 1 (Ntn1), which regulates basal lamina integrity in the fusion plate. Through a series of genetic experiments, we show that mutually antagonistic interactions between Lrig3 and Ntn1 create complementary expression domains that define the future shape of the lateral canal. Remarkably, removal of one copy of Ntn1 from Lrig3 mutants rescues both the circling behavior and the canal malformation. Thus, the Lrig3/Ntn1 feedback loop dictates when and where basement membrane breakdown occurs during canal development, revealing a new mechanism of complex tissue morphogenesis.
- Document Type:
- Reference
- Product Catalog Number:
- AB2034
- Product Catalog Name:
- Anti-Laminin Antibody
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Configuration of a high-content imaging platform for hit identification and pharmacological assessment of JMJD3 demethylase enzyme inhibitors.
- The biological complexity associated with the regulation of histone demethylases makes it desirable to configure a cellular mechanistic assay format that simultaneously encompasses as many of the relevant cellular processes as possible. In this report, the authors describe the configuration of a JMJD3 high-content cellular mechanistic imaging assay that uses single-cell multiparameter measurements to accurately assess cellular viability and the enzyme-dependent demethylation of the H3K27(Me)3 mark by exogenously expressed JMJD3. This approach couples robust statistical analyses with the spatial resolving power of cellular imaging. This enables segregation of expressing and nonexpressing cells into discrete subpopulations and consequently pharmacological quantification of compounds of interest in the expressing population at varying JMJD3 expression levels. Moreover, the authors demonstrate the utility of this hit identification strategy through the successful prosecution of a medium-throughput focused campaign of an 87 500-compound file, which has enabled the identification of JMJD3 cellular-active chemotypes. This study represents the first report of a demethylase high-content imaging assay with the ability to capture a repertoire of pharmacological tools, which are likely both to inform our mechanistic understanding of how JMJD3 is modulated and, more important, to contribute to the identification of novel therapeutic modalities for this demethylase enzyme.
- Document Type:
- Reference
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple
-
The Human Cytomegalovirus Strain DB Activates Oncogenic Pathways in Mammary Epithelial Cells.
- Human cytomegalovirus (HCMV) establishes a persistent life-long infection and increasing evidence indicates HCMV infection can modulate signaling pathways associated with oncogenesis. Breast milk is an important route of HCMV transmission in humans and we hypothesized that mammary epithelial cells could be one of the main cellular targets of HCMV infection.The infectivity of primary human mammary epithelial cells (HMECs) was assessed following infection with the HCMV-DB strain, a clinical isolate with a marked macrophage-tropism. The impact of HCMV-DB infection on expression of p53 and retinoblastoma proteins, telomerase activity and oncogenic pathways (c-Myc, Akt, Ras, STAT3) was studied. Finally the transformation of HCMV-DB infected HMECs was evaluated using soft agar assay. CTH cells (CMV Transformed HMECs) were detected in prolonged cultures of infected HMECs. Tumor formation was observed in NOD/SCID Gamma (NSG) mice injected with CTH cells. Detection of long non coding RNA4.9 (lncRNA4.9) gene was assessed in CTH cells, tumors isolated from xenografted NSG mice and biopsies of patients with breast cancer using qualitative and quantitative PCR.We found that HCMV, especially a clinical strain named HCMV-DB, infects HMECs in vitro. The clinical strain HCMV-DB replicates productively in HMECs as evidenced by detection of early and late viral transcripts and proteins. Following infection of HMECs with HCMV-DB, we observed the inactivation of retinoblastoma and p53 proteins, the activation of telomerase activity, the activation of the proto-oncogenes c-Myc and Ras, the activation of Akt and STAT3, and the upregulation of cyclin D1 and Ki67 antigen. Colony formation was observed in soft agar seeded with HCMV-DB-infected HMECs. Prolonged culture of infected HMECs resulted in the development of clusters of spheroid cells that we called CTH cells (CMV Transformed HMECs). CTH cells when injected in NOD/SCID Gamma (NSG) mice resulted in the development of tumors. We detected in CTH cells the presence of a HCMV signature corresponding to a sequence of the long noncoding RNA4.9 (lncRNA4.9) gene. We also found the presence of the HCMV lncRNA4.9 sequence in tumors isolated from xenografted NSG mice injected with CTH cells and in biopsies of patients with breast cancer using qualitative and quantitative PCR.Our data indicate that key molecular pathways involved in oncogenesis are activated in HCMV-DB-infected HMECs that ultimately results in the transformation of HMECs in vitro with the appearance of CMV-transformed HMECs (CTH cells) in culture. CTH cells display a HCMV signature corresponding to a lncRNA4.9 genomic sequence and give rise to fast growing triple-negative tumors in NSG mice. A similar lncRNA4.9 genomic sequence was detected in tumor biopsies of patients with breast cancer.
- Document Type:
- Reference
- Product Catalog Number:
- 17-408
- Product Catalog Name:
- EZ-Magna ChIP™ A - Chromatin Immunoprecipitation Kit
-
Amicon® Ultra-15 Centrifugal Filter Devices Non IVD User guide
- Document Type:
- User Guide
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple
-
What is the chemical compatibility of Amicon Ultra?…
- Amicon Ultra units are intended for use with biological fluids and aqueous solutions. Please refer to the user guide, catalogue, or contact Technical Support.
- Document Type:
- FAQ
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple
-
Which membranes are available in Amicon Ultra units?…
- Amicon Ultra units are only available with Ultracel regenerated cellulose material. This high recovery/low binding membrane is offered in 3,000, 10,000, 30,000, 50,000 & 100,000 MWCO.
- Document Type:
- FAQ
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple
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Do you have any centrifugal filter units that are pyrogen free?…
- We do not have any centrifugals devices that we claim are pyrogen free. Even the sterile devices could contain pyrogens.
- Document Type:
- FAQ
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple
-
Can the Amicon Ultra be used with nucleic acids?…
- Yes, Amicon Ultra units have been evaluated for use with Genomic and PCR fragments. The 30 kDa MWCO provides the best balance between recovery and spin time. Protocols related to Genomic and PCR fragments can be found in the user guide.
- Document Type:
- FAQ
- Product Catalog Number:
- Multiple
- Product Catalog Name:
- Multiple