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SMD1168, Pichia pastoris Yeast Strain (Invitrogen™)

Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain.

Drosophila S2 Cells in Schneider's Medium (Gibco™)

Gibco® Drosophila S2 cells are used for heterologous protein expression in the DrosophilaM Expression System (DES®). The S2 cell line was derived from a primary culture of late stage (20-24 hours old) Drosophila melanogaster embryos. Many features of the S2 cell line suggest that it is derived from a macrophage-like lineage. S2 cells grow at room temperature in ambient CO2 as a loose, semi-adherent monolayer in tissue culture flasks and in suspension in spinners and shake flasks. Drosophila S2 cells (frozen in Gibco® Schneider's Medium) feature:
• Expression of protein transiently or through generation of stable cell lines
• Quality and performance testing

Expression of protein transiently or through generation of stable cell lines
Drosophila S2 cells can be transfected with the recombinant expression vector alone for transient expression studies or in combination with a selection vector (e.g. pCoHygro or pCoBlast) to generate stable cell lines. A variety of DES® kits are available to allow expression of the recombinant protein of interest in S2 cells.

Quality and performance testing
Each lot of Gibco® S2 cells is tested for cell growth and viability post-recovery from cryopreservation. In addition, the Master Seed Bank has been tested for contamination of bacteria, yeast, mycoplasma and virus and has been characterized by isozyme and karyotype analysis.

Caution: Handle as potentially biohazardous material under at least Biosafety Level 2 containment. This product contains Dimethyl Sulfoxide (DMSO), a hazardous material. Review the Material Safety Data Sheet before handling.

Sf21 cells in Sf-900™ III SFM (Gibco™)

Gibco® Sf21 cells are commonly used to isolate and propagate recombinant baculoviral stocks and to produce recombinant proteins. Gibco® Sf21 cells are adapted to serum-free suspension culture in Gibco® Sf-900™ III SFM, which saves significant time and expense associated with the adaptation of cultures. Gibco® Sf21 cells (frozen in Gibco® Sf-900™ III SFM) feature:
• Recombinant protein expression from a variety of expression systems
• Good growth in adherent or suspension culture
• Quality and performance testing

Recombinant protein expression from a variety of expression systems
High levels of protein expression in Sf21 cells can be obtained using either the BaculoDirect™ Baculovirus Expression System, the Bac-to-Bac® Baculovirus Expression System, or the InsectDirect® Expression System.

Good growth in adherent or suspension culture
Each vial containing 1 × 107 cells can be thawed directly into suspension or adherent culture. Protocols for adherent and suspension growth in Gibco® Sf-900™ III SFM are available in the product manual. Cultures can easily be transferred between the two conditions for ease of workflow but we recommend the stock cultures are maintained in suspension.

Quality and performance testing
Each lot of Gibco® Sf21 cells is tested for cell growth and viability post-recovery from cryopreservation.

Product Use
For Research Use Only. Not for any animal or human therapeutic or diagnostic use. Caution: Handle as potentially biohazardous material under at least Biosafety Level 2 containment. This product contains Dimethyl Sulfoxide (DMSO), a hazardous material. Review the Material Safety Data Sheet before handling.

X-33, Pichia pastoris Yeast Strain (Invitrogen™)

Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain.

Flp-In™-CV-1 Cell Line (Invitrogen™)

Flp-In™ Cell Lines are designed for rapid generation of stable cell lines that express a protein of interest from a Flp-In™ expression vector. These cells contain a single stably integrated FRT site at a transcriptionally active genomic locus. Targeted integration of a Flp-In™ expression vector ensures high-level expression of your gene of interest. There are six Flp-In™ Cell Lines available for the generation of isogenic stable cell lines and a Flp-In™ T-REx™ Cell Line for the generation of tetracycline-regulated stable cell lines. Flp-In™-CV-1, Flp-In™-293, Flp-In™-BHK, Flp-In™-Jurkat, and Flp-In™-3T3 Cell Lines were created by transfecting the parent cell lines with pFRT/lacZeo and selecting for stable Zeocin™- resistant clones. The Flp-In™-CHO Cell Line was created by transfecting CHO cells with pFRT/lacZeo2 and selecting for Zeocin™-resistant clones. The Flp-In™ T-REx™-293 Cell Line contains pFRT/lacZeo and pcDNA™6/TR (from the T-REx™ System) stably integrated. Co-transfection of the Flp-In™ Cell Lines with a Flp-In™ expression vector and the Flp recombinase vector, pOG44, results in targeted integration of the expression vector to the same locus in every cell, ensuring homogeneous levels of gene expression.

Choosing your Flp-In™ Vector/Cell Line Combination
The Flp-In™-CV-1, Flp-In™-293, Flp-In™-CHO, and Flp-In™-Jurkat Cell Lines (Figure 1) work well with Flp-In™ vectors that express a gene from the CMV promoter (pcDNA™5/FRT, pcDNA5/FRT/V5-His-TOPO®, and pSecTag/FRT/V5-His-TOPO®). Flp-In™-BHK and Flp-In™-3T3 cells tend to down regulate the CMV promoter. Therefore, it is recommended that the Flp-In™ vectors containing the EF-1α promoter (pEF5/FRT/V5-DEST™ and pEF5/FRT/V5-D-TOPO®) be used with these cell lines.

GS115, Pichia pastoris Yeast Strain (Invitrogen™)

Several Pichia pastoris strains are available to allow you to optimize expression and recovery of your protein of interest. Table 1 provides information to help you choose the appropriate strain.

INVSc1, S. cerevisiae Yeast Strain (Invitrogen™)

The S. cerevisiae strain, INVSc1, is a fast-growing diploid strain ideal for expression. INVSc1 has the following genotype:

INVSc1: MATa his3D1 leu2 trp1-289 ura3-52 MAT his3D1 leu2 trp1-289 ura3-52

Viral Production Cells (Gibco™)

Gibco Viral Production Cells are a clonal derivative of the HEK 293F cell line, and are a core component of the LV-MAX Lentiviral Production System. Viral Production Cells have been adapted to a special chemically-defined, serum-free and protein-free LV-MAX Production Medium. They are maintained in suspension culture and will grow to high density in LV-MAX Production Medium. Viral Production Cells are highly transfectable by LV-MAX transfection reagent and generate superior lentiviral production compared to polyethyleneimine (PEI)-based lentiviral production methods.

• Suspension culture cells facilitate high-throughput screening or scale-up
• Higher viral production levels than other suspension HEK 293 cells
• Rapid cell growth and high culture viabilities

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in LV-MAX Production Medium and 10% DMSO. The cells should be thawed directly into LV-MAX Production Medium.

The cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable in volume from 96-deep well blocks to 2-L flasks and bioreactors. Lentiviral vector harvest is typically performed at two days post-transfection.

ExpiSf9™ Cells (Gibco™)

ExpiSf9 Cells are a non-engineered derivative of Sf9 insect cells that have been adapted for high-density suspension growth in ExpiSf CD Medium.

ExpiSf9 Cells:
• Enable consistent protein expression performance for over 20 passages after thaw
• Allow you to attain yields of three times more protein per liter than other insect cells from baculovirus-infected culture as part of the ExpiSf Expression System
• Achieve maximum cell densities of 2x107 cells per mL in ExpiSf CD Medium
• Exhibit fast recovery post-thaw and doubling times of approximately 24 hours
• Achieve high performance growth in multiple culture volumes from 4 mL in deep-well plates to >3 L in shake flasks

ExpiSf9 Cells are a core component of the ExpiSf Expression System. Frozen cells are supplied in a vial containing 1.5 mL of cells at 1 x 107 viable cells per mL in ExpiSf CD Medium and 7.5% DMSO. The cells should be thawed in ExpiSf CD Medium. Cells are typically grown in shake flasks on a shaker platform in a non-humidified, non-CO2 cell culture incubator. Growing ExpiSf9 cells in adherent format for routine culture is not recommended.

Human OATP2B1 SLC Transporter Cells (Gibco™)

TRANSiPORT Human OATP2B1 SLC Transporter Cells are HEK293 cells that transiently overexpress the OATP2B1 solute carrier (SLC) transport protein. Solute carriers are a super-family of membrane transporters that can affect pharmacokinetics and drug exposure by governing the transport of solutes in and out of cells. The OATP2B1 SLC transporter protein is found in hepatocytes.

• Assess potential for transporter-mediated drug metabolism
• Easy-to-use format
• Obtain high-quality results with a large signal-to-noise ratio

Choice of measurement system
Cell-based assays can be performed using radioisotope-labeled compounds, fluorescence-labeled compounds, or non-labeled compounds. The amount of substrate transported into the cells can be measured directly using a liquid scintillation counter, fluorescence plate reader, or LC-MS/MS, thereby allowing direct evaluation of SLC transporter activity.

Assay reliability
Mock SLC Transporter Cells (Cat. No. GM1001) that do not overexpress the SLC transporter are available for use as a negative experimental control. Some compounds may demonstrate high background levels of transport due to the presence of endogenous transporters or non-specific binding.

Rapid results
The convenient product format enables data generation in two days from thawing of the cells to final results.

Flp-In™ T-REx™ 293 Cell Line (Invitrogen™)

Flp-In™ 293 T-REx cell lines is designed for rapid generation of stable cell lines that ensure homogenous expression of your protein of interest from a Flp-In™ expression vector. These cells contain a single stably integrated FRT site at a transcriptionally active genomic locus. Targeted integration of a Flp-In™ expression vector ensures high-level expression of your gene of interest. The Flp-In™ T-REx™-293 Cell Line contains pFRT⁄lacZeo and pcDNA™6⁄TR (from the T-REx™ System) stably integrated. Co-transfection of the Flp-In™ Cell Lines with a Flp-In™ expression vector and the Flp recombinase vector, pOG44, results in targeted integration of the expression vector to the same locus in every cell, ensuring homogeneous levels of gene expression.

The Flp-In™-293 work well with Flp-In™ vectors that express a gene from the CMV promoter (e.g., pcDNA™5⁄FRT, pcDNA5⁄FRT⁄V5-His-TOPO®, and pSecTag⁄FRT⁄V5-His-TOPO®).

Expi293F™ GnTI- Cells (Gibco™)

The Expi293F GnTI- cell line is derived from engineered Expi293F cells that do not have N-acetylglucosaminyltransferase I (GnTI) activity and therefore lack complex N-glycans. It is a powerful tool for the high-yield expression of homogeneously glycosylated recombinant proteins.

Features of Expi293F GnTI- cells include:
• Suspension, high-density culture in Expi293 Expression Medium
• Homogeneous N-glycosylation of expressed proteins
• Equivalent protein yields to parental Expi293F cells—up to 1 g/L
• Significantly higher protein yields than other HEK293 GnTI- cell line alternatives

Frozen cells are supplied in a vial containing 1 mL of cells at 1 x 107 viable cells/mL in Expi293 Expression Medium and 10% DMSO. The cells should be thawed directly into Expi293 Expression Medium.

Expi293F™ Cells (Gibco™)

Expi293F™ human cells are derived from the 293 cell line, and are a core component of the Expi293™ Expression System. They are maintained in suspension culture and will grow to high density in Expi293™ Expression Medium. Expi293F™ cells are highly transfectable and generate superior protein yields compared to standard 293 cell lines in transient protein expression.

Expi293F Cells attributes include:
Suspension culture cells pre-adapted to Expi293™ Expression Medium
Demonstrated higher protein expression levels than other 293 cells adapted to Expi293™ Expression Medium
Exhibit more rapid cell growth and higher culture viabilities than standard suspension-adapted 293 cells
Consistent transient expression performance of Expi293F™ cells for over 40 passages beyond thaw
• Also available from a cGMP bank (Cat. No. 100044202)

Frozen cells are supplied in a vial containing 1 ml of cells at 1 x 107 viable cells/ml in Expi293™ Expression Medium and 10% DMSO. The cells should be thawed directly into Expi293™ Expression Medium. The six-vial multipack eliminates the need to expand and rebank the cells in the lab.

Cells are typically grown in flasks on a shaker platform in a humidified, CO2 cell culture incubator. However, cultures are scalable from volumes of <1 mL in multiwell plates to >10 L in bioreactors. Protein harvest can be performed at 2 to 7 days post-transfection, depending on the protein expressed. For expression of recombinant antibodies, harvest is typically at 5 to 7 days post-transfection. There is no need for culture medium supplementation or batch feeding during expression.

Flp-In™-CHO Cell Line (Invitrogen™)

Flp-In™ Cell Lines are designed for rapid generation of stable cell lines that express a protein of interest from a Flp-In™ expression vector. These cells contain a single stably integrated FRT site at a transcriptionally active genomic locus. Targeted integration of a Flp-In™ expression vector ensures high-level expression of your gene of interest. There are six Flp-In™ Cell Lines available for the generation of isogenic stable cell lines and a Flp-In™ T-REx™ Cell Line for the generation of tetracycline-regulated stable cell lines. Flp-In™-CV-1, Flp-In™-293, Flp-In™-BHK, Flp-In™-Jurkat, and Flp-In™-3T3 Cell Lines were created by transfecting the parent cell lines with pFRT/lacZeo and selecting for stable Zeocin™- resistant clones. The Flp-In™-CHO Cell Line was created by transfecting CHO cells with pFRT/lacZeo2 and selecting for Zeocin™-resistant clones. The Flp-In™ T-REx™-293 Cell Line contains pFRT/lacZeo and pcDNA™6/TR (from the T-REx™ System) stably integrated. Co-transfection of the Flp-In™ Cell Lines with a Flp-In™ expression vector and the Flp recombinase vector, pOG44, results in targeted integration of the expression vector to the same locus in every cell, ensuring homogeneous levels of gene expression.

Choosing your Flp-In™ Vector/Cell Line Combination
The Flp-In™-CV-1, Flp-In™-293, Flp-In™-CHO, and Flp-In™-Jurkat Cell Lines (Figure 1) work well with Flp-In™ vectors that express a gene from the CMV promoter (pcDNA™5/FRT, pcDNA5/FRT/V5-His-TOPO®, and pSecTag/FRT/V5-His-TOPO®). Flp-In™-BHK and Flp-In™-3T3 cells tend to down regulate the CMV promoter. Therefore, it is recommended that the Flp-In™ vectors containing the EF-1α promoter (pEF5/FRT/V5-DEST™ and pEF5/FRT/V5-D-TOPO®) be used with these cell lines.

T-REx™-CHO Cell Line (Invitrogen™)

T-REx™ Cell Lines stably express the tetracycline repressor protein (Table 1). They save significant time and effort when using the T-REx™ System. The T-REx™ Cell Lines are functionally tested by transient transfection with the positive control vector pcDNA™4⁄TO⁄lacZ. T-REx™ Cell Lines exhibit extremely low basal expression levels in the repressed state and high expression upon induction with tetracycline (Figure 1).