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Neoplastic transformation occurs via a series of genetic and epigenetic alterations that yield a cell population that is capable of proliferating independently of both external and internal signals that normally restrain growth. Anchorage-independent growth is one of the hallmarks of cell…
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Using adenovirus to deliver genes to target cells is an option for researchers who are interested in a high titer virus with high transient expression levels that infects a wide range of cell types. Although adenovirus does not integrate into the genome of the target cell the way a retrovirus…
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Recombinant adeno-associated virus (AAV) is often the preferred method for delivering genes to target cells due to its high titer, mild immune response, ability to infect a broad range of cells, and overall safety. Native AAV can infect humans and primates; however it has not been reported to…
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Q: Can this be used with cells other than HUVECs?
A: This assay can be used with any endothelial cells.
Q: What is the composition of the ECM gel solution?
A: The ECM gel solution is the same as matrigel, which is an ECM extract prepared from Engelbreth-Holm-Swarm (EHS) sarcoma produced in mice…
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Q: What is the assay principle for your ECM Cell Adhesion Assays?
A: Our Cell Adhesion Assays use an ECM coated plate for cell seeding. The wells are then washed with PBS to remove any non-adherent cells. The remaining adherent cells are stained and the dye is then extracted for…
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Q: How do the Microfluidic Biochips work for studying cell adhesion?
A: This product is designed to measure cell adhesion in a microfluidic environment that resembles in vivo shear stresses. The protocol involves coating the biochip with your desired cell adhesion molecule, adding endothelial…
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Q: How is the gel released from the well?
A: After a two day culture, a sterile spatula is carefully inserted between the plate wall and the gel. The spatula is then slowly moved around the wall in a circle to completely release the gel. We use a small sterile spatula with a thin, flat, edge.
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Q: Can the plates in your Anoikis Assays be re-used?
A: Our Anoikis assay uses a hydrogel coated plated that cells cannot attach to, which promotes anoikis cell death. The hydrogel integrity on the surface of the plate may not be maintained throughout the assay protocol and we don’t recommend…
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Q: Can this assay be used with collagen I coated plates?
A: It is fine to culture cells on any type of plate with the CBA-240 assay. Culturing cells on collagen-I will not affect the assay.
Q: Can this assay be used with bacterial cells?
A: Our Cell Viability and Cytotoxicity Assay (…
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Q: Can any of your Cellular Senescence Assays be automated?
A: For automating the senesce assay screening, we recommend using #CBA-231, which comes in a 96-well format and can be read with a fluorescent plate reader. This is an activity assay that quantitates senescence by measuring SA-ß-gal…
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Q: Is this assay compatible with tissue samples?
A: Yes this kit can be used to stain tissue samples, and no modifications are necessary to the protocol.
Q: I did not detect a blue color with this kit.
A: The absence of blue color with the senescence assay indicates that there are few…
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Q: How is SA-ß-Gal activity measured with this assay?
A: This assay uses a fluorometric substrate to detect SA-ß-Gal enzyme activity in cell lysates. The buffers included in this kit maintain pH 6.0 throughout the assay to ensure optimal conditions for measuring SA-ß-Gal while suppressing…
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Q: What is the purpose of the Pretreatment Solution?
A: There are two main types of β-galactosidases in senescent cells: SA-β-galactosidase and Lysosomal β-galactosidase. Lysosomal β-galactosidase activity can only be detected at pH 4. The Cell Pretreatment Solution is used to induce…
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Q: Are your Cell Transformation Assay Kits used the same way as a traditional soft agar assay?
A: Our 96-well Cell Transformation Assays are used in a similar manner as the traditional soft agar assay, which measures anchorage independent growth. The main advantages to our kits are that they…
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Q: Is it acceptable to add antibiotic to the medium?
A: Antibiotics can be used in the media with this assay, and it will be able to diffuse through the cell agar layer. Any media that is regularly used can be used with this assay, as long as it is 2X.
Q: Does the media need to be…
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Q: You say manual counting of cells is eliminated when using your Cell Transformation Kits with Cell Recovery, but how can I quantify my cells and still recover them intact for further analysis?
A: All of our Cell Transformation Assays include a quantitation step that eliminates manual cell…
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Q: Which Phagocytosis Assay should I use?
A: The selection of the appropriate kit will be determined based on the desired phagocytosis substrate. Our phagocytosis assays use opsonized RBCs (#CBA-220), Zymosan particles (#CBA-224), or E.coli (#CBA-222) as a phagocytosis substrate.
Q: What…
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Q: Is this kit compatible with any cell line?
A: Our Cell Transformation Assays, including our In Vitro Tumor Sensitivity Assay, are not cell type specific and can be used for any cell line as long as the cells can form colonies in soft agar.
Q: Can any media be used with this assay?…
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Q: What size should the tumors be prior to mincing?
A: The size of the tumor used in the assay is not critical because there is a step that adjusts for cell number (step 14 of the Assay Protocol found in the product manual). It is difficult to recommend a size because the cell density will…
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Q: I have never run an assay using a Boyden Chamber. Would I have a better chance of success with your Cell Invasion kits if I use the 24-well or 96-well format?
A: If you are in the early stages of your invasion studies and are still optimizing conditions, it is recommended that you start with the…
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Q: Is the colorimetric or fluorometric kit more sensitive?
A: The fluorometric assay is more sensitive than the colorimetric assay, however manual counting can be performed with the colorimetric assay following the staining step and prior to the extraction step. Manual counting is always more…
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Q: Does the entire 96-well Invasion Chamber plate need to be used in one experiment?
A: Yes the 96-well invasion assay must be used all at once. Since all the coated inserts are in a single plate, they will all have to go through the overnight incubation of the invasion assay in a 37ºC…
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Q: How is chemotaxis measured using a Boyden Chamber system?
A: When using a Boyden Chamber, the cells are in serum-free medium in the upper chamber and the media with the chemoattractant is in the lower chamber, which establishes a chemoattractant concentration gradient between the two…
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Q: Can the inserts be reused?
A: Our 24-well migration assays contain 12 individual, removable inserts made out of polycarbonate. Any unused inserts can be stored at 4ºC until needed, however the inserts should not be used more than once.
Q: Can the inserts be stored after…