An article published this year in “Analytical and Bioanalytical Chemistry” using our “Polyclonal Antibody Service”, by our customers from Faculty of ChemistryComplutense University of Madrid, Humanitas Clinical and Research Center, Rozzano, Italy, in the analysis of how MicroRNA-26a/cyclin-dependent kinase 5 axis controls proliferation, apoptosis and in vivo tumor growth of diffuse large B-cell lymphoma cell lines. Congrats and Thanks.

Summay:

The assembly of a novel disposable amperometric immunosensor for the detection of the red wine spoilage yeast Brettanomyces bruxellensis is reported. The nanostructured sensing interface was prepared by first coating carbon screen printed electrodes with a gold nanoparticles-reduced graphene oxide hybrid nanomaterial, which was then modified with 3-mercaptopropionic acid to further immobilize specific antibodies for B. bruxellensis via a carbodiimide-coupling reaction. The functionalized electrode allowed the amperometric detection of B. bruxellensis in buffered solutions and red wine samples in the range of 10–10⁶ CFU/mL and 10²–10⁶ CFU/mL, with low detection limits of 8 CFU/mL and 56 CFU/mL, respectively. The electrochemical immunosensor also exhibited high reproducibility, selectivity, and storage stability.

Reference:

https://link.springer.com/article/10.1007%2Fs00216-017-0505-5

Product link:

Immunostep Polyclonal Antibody Service

http://www.immunostep.com/content/27-monoclonal-and-polyclonal-antibody-development-service

CFBlue Annexin V Apoptosis Detection Kit with PI

An article published this year in “NANOSCALE” using our CFBlue Annexin V Apoptosis Detection Kit with PI, by our customers from Department of Medicine and General Cytometry Service-Nucleus, Cancer Research Centre (IBMCC/CSIC/USAL/IBSAL), Salamanca, Spain., in the analysis of Functional insights into the cellular response triggered by a bile-acid platinum compound conjugated to biocompatible ferric nanoparticles using quantitative proteomic approaches.. Congrats and Thanks.

Summary:

At present, bioferrofluids are employed as powerful multifunctional tools for biomedical applications such as drug delivery, among others. The present study explores the cellular response evoked when bile-acid platinum derivatives are conjugated with bioferrofluids by testing the biological activity in osteosarcoma (MG-63) and T-cell leukemia (Jurkat) cells. The aim of this work is to evaluate the biocompatibility of a bile-acid platinum derivative conjugated with multi-functional polymer coated bioferrofluids by observing the effects on the protein expression profiles and in intracellular pathways of nanoparticle-stimulated cells. To this end, a mass spectrometry-based approach termed SILAC has been applied to determine in a high-throughput manner the key proteins involved in the cellular response process (including specific quantitatively identified proteins related to the vesicular transport, cellular structure, cell cycle, biosynthetic process, apoptosis and regulation of the cell cycle). Finally, biocompatibility was evaluated and validated by conventional strategies also (such as flow cytometry, MTT, etc.).

Reference:

http://pubs.rsc.org/en/Content/ArticleLanding/2017/NR/C7NR02196H#!divAbstract

Product link:

CFBlue Annexin V Apoptosis Detection Kit with PI: http://www.immunostep.com/apoptosis-tools/3735-anxvkcfb-100t.html

Annexin-V-DY634 and 7AAD staining.

An article published this year in “OSTEOARTHRITIS AND CARTILAGE” using our annexin-V-DY634 and 7AAD staining, by our customers from Departamento de Bioquímica, Biología Molecular y Celular, Universidad de Zaragoza, and Servicio de Inmunología Hospital Clínico Universitario Lorenzo Blesa, Zaragoza, Spain, in the analysis of how CD56+/CD16- Natural Killer cells expressing the inflammatory protease granzyme A are enriched in synovial fluid from patients with osteoarthritis. Congrats and Thanks.

Summary:

Objective: NK cells have been involved in the pathology of different inflammatory and autoimmune disorders. Inflammation is an important regulator of osteoarthritis (OA), but the molecular and cellular mechanisms regulating this process are not well defined.

Design: To understand the role of NK cells in OA, we have compared the phenotype (CD56 subsets and perforin and granzyme expression) and cytotoxic function of NK cells in peripheral blood and synovial fluid from patients with OA undergoing total knee arthroplasty.

Results: In contrast to peripheral blood lymphocytes, the majority of NK cells from the synovial fluid were CD56brightCD16(-) cells. As expected the expression of the cytolytic mediators perforin and granzyme B in CD56brightCD16(-) cells was low and correlated with a poor cytotoxic potential against K562 sensitive target cells. Surprisingly, this low cytotoxic NK cell subset (but not in peripheral blood) expressed high levels of granzyme A (a protease recently characterized as a key modulator of inflammation in mouse models) in synovial fluid but not in peripheral blood. The presence of the CD56(+)brightCD16(-) cells expressing granzyme A correlated with increased levels of pro-inflammatory cytokines in synovial fluid from OA patients.

Conclusion: Our results indicate that NK cells from the synovium of patients with OA, which present an immunoregulatory non-cytotoxic phenotype, show different phenotype comparing with NK cells from peripheral blood, especially expressing granzyme A, a pro-inflammatory molecule which may contribute to the establishment of chronic articular inflammation in this type of patients.

Reference:

http://www.sciencedirect.com/science/article/pii/S1063458417310555

Product link:

DY634 Annexin V: http://www.immunostep.com/apoptosis-tools/3730-anxvdy-200t.html

7-AAD Staining Solution: http://www.immunostep.com/solutions-buffers-chemicals/2307-7-aad-staining-solution.html

An article published this year in “PEERJ” using our propidium iodide/RNAse solution, by our customers from Department of Cellular and Molecular Biology, Centro de Investigaciones Biológicas, Spanish National Research Council (CSIC), Madrid, Spain, in the analysis of how Differential gene expression analysis by RNA-seq reveals the importance of actin cytoskeletal proteins in erythroleukemia cells. Congrats and Thanks.

Summary:

Development of drug resistance limits the effectiveness of anticancer treatments. Understanding the molecular mechanisms triggering this event in tumor cells may lead to improved therapeutic strategies. Here we used RNA-seq to compare the transcriptomes of a murine erythroleukemia cell line (MEL) and a derived cell line with induced resistance to differentiation (MEL-R). RNA-seq analysis identified a total of 596 genes (Benjamini–Hochberg adjusted p-value < 0.05) that were differentially expressed by more than two-fold, of which 81.5% (486/596) of genes were up-regulated in MEL cells and 110 up-regulated in MEL-R cells. These observations revealed that for some genes the relative expression of mRNA amount in the MEL cell line has decreased as the cells acquired the resistant phenotype. Clustering analysis of a group of genes showing the highest differential expression allowed identification of a sub-group among genes up-regulated in MEL cells. These genes are related to the organization of the actin cytoskeleton network. Moreover, the majority of these genes are preferentially expressed in the hematopoietic lineage and at least three of them, Was (Wiskott Aldrich syndrome), Btk (Bruton’s tyrosine kinase) and Rac2, when mutated in humans, give rise to severe hematopoietic deficiencies. Among the group of genes that were up-regulated in MEL-R cells, 16% of genes code for histone proteins, both canonical and variants. A potential implication of these results on the blockade of differentiation in resistant cells is discussed.

Reference:

https://peerj.com/articles/3432/

Product link:

CELL CYCLE ANALYSIS (PI / RNASE): http://www.immunostep.com/60-cell-cycle-analysis-pi-rnase

Annexin-V–Allophycocyanin (APC)/Propidium Iodide (PI) double staining.

An article published this year in “CELL DEATH AND DISEASE” using our Annexin-V–Allophycocyanin (APC)/Propidium Iodide (PI) double staining, by our customers from Humanitas Clinical and Research Center, Rozzano, Italy, in the analysis of how MicroRNA-26a/cyclin-dependent kinase 5 axis controls proliferation, apoptosis and in vivo tumor growth of diffuse large B-cell lymphoma cell lines. Congrats and Thanks.

Summary:

Diffuse large B-cell lymphoma (DLBCL) is the most frequent type of non-Hodgkin lymphoma. Despite a favorable therapeutic response to first-line chemo-immunotherapy, still 30–40% of patients is refractory, or relapse after this treatment. Thus, alternative strategies must be sought. Previous studies have indicated that cyclin-dependent kinase 5 (CDK5), a serine/threonine protein kinase, is involved in tumor development and progression, and it may represent a potential therapeutic target. However, its role in modulating DLBCL growth and progression remains largely unexplored. In this study, we show that CDK5 and its activator, cyclin-dependent kinase 5 activator 1 (CDK5R1 or p35), are overexpressed in DLBCL cell lines and that signal transducer and activator of transcription 3 (STAT3) phosphorylation and activity is dependent on CDK5 expression in DLBCL. Using public data sets, we also demonstrate that patients with DLBCL show a higher expression of CDK5 compared with healthy individuals. By using loss-of-function approaches, we demonstrate that CDK5’s activity regulates proliferation and survival of DLBCL cells. MicroRNAs (miRNAs or miRs) are small noncoding RNAs that negatively regulating gene expression and are involved in cancer initiation and progression. We identify miR-26a as direct regulator of p35 expression and CDK5 activity. We show that miR-26a expression is lower in DLBCL cell lines compared to B lymphocytes and that its ectopic expression leads to a drastic reduction of DLBCL tumor growth in vivo and decreased proliferation, cell-cycle progression, and survival in vitro. Remarkably, concomitant overexpression of a 3′-UTR-truncated form of p35 promoted tumor growth in vivo and cell proliferation, cell-cycle progression, and cell survival in vitro. In conclusion, these results demonstrate an important role for miR-26a and CDK5 together in the survival and growth of DLBCL cells, suggesting the existence of potential novel therapeutic targets for the treatment of DLBCL.

Reference:

https://www.nature.com/cddis/journal/v8/n6/full/cddis2017291a.html

Product link:

Annexin-V–Allophycocyanin (APC)/Propidium Iodide (PI) double staining: http://www.immunostep.com/57-annexin-v-anxv