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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…
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Q: What is the detection limit with the CyQuant® dye?
A: The limit of detection with CyQuant® is about 500-1000 cells, which is less sensitive than manual counting. Our recommendation to get the highest level of detection is to seed with the maximum number of cells (500,000) in serum free…
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Q: What is the difference between your product #CBA-101-COL where collagen is pre-coated on the underside (bottom) of the membrane, and your product #CBA 111-COL where collagen is coated on top of the membrane?
A: These products assay for different processes – haptotaxis and cell invasion,…
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Q: What endothelial cells are compatible with this assay?
A: We use HUVEC cells, but any endothelial cells can be used.
Q: How can I confirm that the insert is evenly covered with endothelial cells?
A: After culturing the endothelial cells on the insert for 2-3 days, the insert can be…
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Q: How does the Radius™ assay work?
A: Our Radius assay uses a plate that contains a biocompatible hydrogel at the center of the well; seeded cells will attach everywhere but on the hydrogel. After cells attach, the gel is removed by addition of a removal solution and the assay begins.
Q…
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Q: What cell lines are compatible with this assay?
A: Our Wound Healing Assay is compatible with any adherent cell line.
Q: Can the inserts be used with a different plate?
A: We don’t recommend using a different plate with the inserts. The wound inserts are custom made with a plastic…
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Q: I want to test my samples for oxidative stress, but there are so many oxidative stress markers. How do I know which assays to use?
A: Oxidative stress may be tested either directly, by measuring the presence of various reactive oxygen species, or indirectly by measuring the resulting damage…
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Sample preparation
Q: Do I need to extract DNA if using urine, plasma, or serum?
A: It is not necessary to extract DNA if samples are a body fluid such as urine, plasma, serum, cerebrospinal fluid, or saliva. Samples may need to be briefly spun down if there are insoluble particles in the sample,…
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Q: Does this antibody also detect 8-OHdG?
A: The primary antibody used in our 8-OHG kit is monoclonal and will also react with 8-OHdG. We recommend using purified RNA samples to specifically detect 8-OHG. If using urine, serum or cerebrospinal fluid samples, both 8-OHdG and 8-OHG will be…