Altogen Custom Research Services: Biology CRO

Generation of Stably Expressing Cell Lines in 28 Days

Altogen Labs offer generation of stable cell lines by transforming your cell line to stably express vector or gene of interest. Plasmid DNA construct should encode at least one antibiotic resistance gene to enable drug selection. Transfected genetic material can be expressed in the target cells either transiently or permanently depending on the methods utilized and the experimental questions being investigated. Transient transfections are used most commonly to analyze the short term impact of altered gene or protein expression. Plasmid DNA (pDNA), messenger RNA (mRNA), short interfering RNA (siRNA), and microRNA (miRNA), are introduced and gene products are expressed in the target cells however the nucleic acids do not integrate into the host cell genome. Therefore, gene product expression is transient and typically results in high expression levels that persist for 24-72 hours when RNA is transfected, or 48-96 hours following DNA transfection. Conversely, in order to analyze the long term impact of altered gene or protein expression investigators typically utilize stable transfection protocols to develop stable cell lines. In a subpopulation of transfected cells, whether the desired effect is a stable or transient transfection, the transfected genetic material will integrate into the genome. In order to create stable cell lines, investigators will take advantage of this natural occurrence, and introduce the gene of interest along with a selectable marker. Therefore growth of transfected cells, in the presence of a selecting agent, will enable the subpopulation of cell where the exogenous genetic material has been incorporated into the genome to persist while the remaining cells undergo selection. Utilizing this method, investigators are able to develop cells that permanently express specific genes through their incorporation in the cellular genome.

For more information please contact us at techserv@altogen.com Experimental details will help us to provide an accurate quote.

Generation of Stable Cell Lines in 28 Days

RNAi Services

RNA interference (RNAi) is a phenomenon by which the expression of double stranded RNA (dsRNA) specifically stimulates a cellular process that reduces gene expression in a sequence specific manner. Small synthetic RNA, termed small interfering RNAs (siRNA), which are typically 21-28 nucleotides in length, can induce RNAi and knockdown gene expression in mammalian cells without inducing an antiviral response. Gene silencing by RNA Interference (RNAi) is a powerful research tool for studying gene function in mammalian cells. RNAi is a biological phenomenon by which double stranded RNA (dsRNA) specifically reduces gene expression of its corresponding gene. Potent inhibition of specific gene expression is experimentally achieved by the transfection of small interfering RNA (siRNA), which binds RISC complex and cause degradation of target complementary mRNA molecules in the cell. Therefore, successful, potent RNAi experimentation is dependent upon the highly efficient delivery of the siRNA into cells by transfection of stable and functional siRNA molecules. Altogen Labs offer following RNAi Services:

  • siRNA transfection optimization service
  • Design and synthesis of siRNA targeting gene of interest
  • Custom siRNA assay development, siRNA library screening
  • Gene of interest knockdown service

Please contact us at techserv@altogen.com for more information. Experimental details will help us to provide an accurate quote.

RNAi Services

MTS/PMS In Vitro Cytotoxicity Assays and IC50 for Tumor Cell Lines

Cytotoxicity Assay

Cytotoxicity assays are based on the cleavage of the yellow terazolium salt (XTT) by metabolically active human cells (HeLa) to form an orange formazan dye. This conversion only occurs in viable cells, so cytotoxicity leads to a loss of activity. The formazan dye formed is soluble in aqueous solutions and is directly quantified using a scanning multi-well spectrophotometer. This method ensures a high degree of accuracy and enables on-line computer processing of the data. Cell viability is calculated as a percentage of standard controls. The assay is suitable for the performance of dose-response studies and is offered in a 96-well microplate format.

MTS/PMS In Vitro Cytotoxicity Assay

This service determines the effect of a compound of interest on the proliferation of one or more tumor cell lines; an MTS/PMS colorimetric assay is used to estimate proliferating cell number per well. Cell proliferation is calculated as a percentage of standard controls. The assay is suitable for the performance of dose-response studies and is offered in a 96-well microplate format.

IC50 for Tumor Cell Lines

This service is designed to measure the IC50 of a compound of interest in over 70 cancer cell lines. Cells in logarithmic growth are seeded in 96-well microtiter plates (5000 cells/well). Control conditions are (1) cells with medium alone and (2) cells with medium and vehicle. Plates are assayed at 48-72 hr after initiation of compound exposure, then a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) solution is added to each well for a 2- to 4-h incubation (37°C, 5% CO2). Analysis is performed in triplicate using a plate reader, and the IC50 values are calculated.

Please contact us at techserv@altogen.com for more information. Experimental details will help us to provide an accurate quote.

CRO Services by Altogen Biosystems

Phosphodiesterase, Phosphatase, Kinase, and ELISA Assay Development

Altogen Labs offers kinase, phosphatase, and phosphodiesterase assay development services for screening and validation studies. For each project, an individual pathway is defined that matches the biology and chemistry requirements of the target to the most appropriate assay technology platform available. Company scientists have experience developing assays with all major fluorometric readouts and biologically relevant formats, in addition to high-content applications: fluorescence, time-resolved fluorescence resonance energy transfer, luminescence, absorbance, calcium mobilization (FLIPR), ELISA, cellular imaging, fluorescent microscopy, enzyme assays, reporter assays, viability assays, and protein translocation. Compound profiling is supported by qRT-PCR, Western blot, FACS and other expression and functional assays.

Please contact us at techserv@altogen.com for more information. Experimental details will help us to provide an accurate quote.

Assay Development Services

Immunohistochemical Staining (IHC Assay Development Service)

Altogen Labs immunohistochemical staining services are available for formalin-fixed, fresh, untrimmed tissue samples, paraffin-embedded tissue blocks, specimens in preservative, or slides (unstained). Company scientists process the specimens and gross to cassette , embed the preserved specimens in paraffin blocks, section the blocks using a microtome, treat the frozen sections with OCT compound and further section the samples with a cryostat to yield 3-µm specimens, and stain the slides with hematoxylin and eosin. Altogen Labs offers optimization of experimental conditions for the primary antibody, including concentrations and incubation times, interpretation of the immunostaining results, and slide staining. Company scientists offer cell line slides and tissues from a variety of organs.

Please contact us at techserv@altogen.com for more information. Experimental details will help us to provide an accurate quote.

Preclinical Research Services

Custom Preclinical Research Services

Custom laboratory research services include:

  • Transfection services (transient and stable)
  • Assay development and library screening
  • Cloning and plasmid vector construction
  • Generation of stable cell lines
  • RNAi experiments (siRNA design, synthesis, validation)
  • Gene targeting (mRNA knockdown, shRNA overexpression)
  • Gene of interest knockdown service
  • Custom services

Altogen Labs provides specialized biotechnology and pharmaceutical services, including generation of stable cell lines, gene silencing and RNA interference services, microorganism identification (bacteria 16S ID), pre-clinical toxicology IND CRO services, in vivo xenograft models, ELISA assay development, siRNA library screening and transfection services, bioremediation products and services.

For more information please contact us at techserv@altogen.com Experimental details will help us to provide an accurate quote.
 

Featured transfection products from Altogen Biosystems:

In Vivo Transfection Kits || Fibroblast Cell Transfection Kit || HepG2 Cell Transfection Reagent || MDA-MB Transfection Reagent || A549 Cell Transfection Reagent || HUVEC Transfection Reagent || LNCaP Cell Transfection Kit ||  In Vivo Nanoparticle Transfection Kit  ||  In Vivo PEG-Liposome Transfection Kit  ||  In Vivo Lipid Transfection Kit  ||  In Vivo Polymer Transfection Kit 

Transfection Resource:

Visit Altogen's Transfection Resource: RNAi applications, stable and transient transfection, delivery methods, electroporation, nanoparticle, microinjection, dendrimer-based, non-viral and virus-mediated gene delivery, liposome and non-liposomal reagents, In Vivo Transfection Reagents from Altogen Biosystems.