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Invitrogen™ KDalert™ GAPDH Assay Kit
Descripción
The Ambion™ KDalert™ GAPDH Assay Kit is for the reliable measure of GAPDH enzyme activity in cultured human, mouse, or rat cells in less than 30 minutes using a microplate fluorometer. The kit includes sufficient reagents for 375 reactions.
Assess GAPDH siRNA delivery in 1/3 the time for 1/3 the cost of real-time PCR
Analyze 1-96 samples simultaneously
Measure both GAPDH siRNA-induced knockdown AND transfection-induced toxicity
Compatible with a wide variety of cells and a broad range of culture conditions
The KDalert GAPDH Assay Kit is an ideal positive control for transfection optimization experiments and also measures transfection induced cytoxicity. It is designed for use with Ambion™ Silencer™ GAPDH siRNA.
Rapid, Time-Saving Procedure
Use the assay to optimize siRNA transfection by transfecting individual cell samples with a GAPDH siRNA and a negative control siRNA. Two to three days after transfection, simply add the included cell lysis buffer to the cells, incubate for 20 minutes, add the diluted master mix of assay reagents, and read the increase in fluorescence four minutes later using a microplate or standard fluorometer. The assay procedure can be completed in about 30 minutes with minimal sample handling.
One Assay for Two Readouts
Because GAPDH is expressed at relatively constant levels, the assay can also be used to monitor transfection agent induced toxicity. For this analysis, GAPDH enzyme activity from negative control siRNA-transfected cells is compared to that of untreated cells. Reduced GAPDH activity in negative control-transfected cells compared to non-transfected cells is an indication that the transfection-induced cytotoxicity.
Accessory Products:
The KDalert™ Kit is designed for use with Silencer™ GAPDH siRNAs (SKUs #AM4605, AM4633, AM4634, AM4624, AM4632, or AM4631). Additional KDalert™ Lysis Buffer (SKU #AM8790G) is also available separately.
The KDalert GAPDH Assay Kit is an ideal positive control for transfection optimization experiments and also measures transfection induced cytoxicity. It is designed for use with Ambion™ Silencer™ GAPDH siRNA.
Rapid, Time-Saving Procedure
Use the assay to optimize siRNA transfection by transfecting individual cell samples with a GAPDH siRNA and a negative control siRNA. Two to three days after transfection, simply add the included cell lysis buffer to the cells, incubate for 20 minutes, add the diluted master mix of assay reagents, and read the increase in fluorescence four minutes later using a microplate or standard fluorometer. The assay procedure can be completed in about 30 minutes with minimal sample handling.
One Assay for Two Readouts
Because GAPDH is expressed at relatively constant levels, the assay can also be used to monitor transfection agent induced toxicity. For this analysis, GAPDH enzyme activity from negative control siRNA-transfected cells is compared to that of untreated cells. Reduced GAPDH activity in negative control-transfected cells compared to non-transfected cells is an indication that the transfection-induced cytotoxicity.
Accessory Products:
The KDalert™ Kit is designed for use with Silencer™ GAPDH siRNAs (SKUs #AM4605, AM4633, AM4634, AM4624, AM4632, or AM4631). Additional KDalert™ Lysis Buffer (SKU #AM8790G) is also available separately.
Especificaciones
Especificaciones
| Content And Storage | KDalert Lysis Buffer can be stored at –20°C or 4°C. Solution A, Solution B, and Solution C should be stored at –20°C. GAPDH Enzyme should be stored at or below –70°C. |
| Assay | GAPDH Assay |
| Detection Method | Fluorescence |
| Product Line | Ambion, KDalert |
| Product Type | GAPDH Assay Kit |
| Substrate Properties | Chemical Substrate |
| Sufficient For | 375 Reactions |
| For Use With (Application) | Absorbance, Fluorescence Intensity |
| Quantity | 375 Reactions |
| Shipping Condition | Dry Ice |
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Frequently Asked Questions (FAQs)
I am using the KDalert GAPDH Assay Kit and am not seeing any GAPDH activity in my standard curve dilutions. What is the problem?
If none of the standard curve samples turn a magenta color after 15 minutes, there was most likely an error in preparing the KDalert Master Mix. We suggest repeating the GAPDH Enzyme dilution standard curve reactions.
For Research Use Only. Not for use in diagnostic procedures.
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