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In the era of high-content screening, scientists are trying to identify rapid, automated formats for immunosorbent assays (ELISA). The ANTI-FLAG High Sensitivity 96-well plates provide a convenient, high-throughput platform for the capture and detection of recombinant FLAG fusion proteins (Figures 1 and 2). The ANTI-FLAG® High Sensitivity plates are coated with the mouse monoclonal M2 antibody and pre-blocked to provide timesavings for high-throughput users. The M2 monoclonal antibody is covalently bound to the plate in a favorable orientation such that the Fab region of the antibody is available for the epitope tag to provide greater specificity. The ANTI-FLAG coating can detect as little as 1 ng/well with a capacity of up to 300 ng/well of FLAG fusion protein.
An automated method for high-throughput ELISA applications has been developed and validated for the ANTI-FLAG High Sensitivity plates. The following resources are available for download: Automated Method and Automated Protocol for use with the Sciclone ALH 3000 Workstation from Caliper Life Sciences.
Applications
- Screening for expression
- Protein: protein interaction assays
Features
Rapid Procedure
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96 samples can be processed from cell lysis through recombinant protein purification in 4-5 hours.
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Enhanced Productivity
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Walk-away automation of recombinant FLAG fusion protein capture and detection by ELISA allows you to concentrate on other aspects of your work.
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Simplified Handling
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The optimized, pre-blocked M2 monoclonal antibody coating eliminates several time-consuming steps associated with traditional ELISA experiments.
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High Sensitivity
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The ANTI-FLAG coating can detect as little as 1 ng/well of FLAG fusion protein.
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Order Information
ANTI-FLAG High Sensitivity M2 Coated Plates
| Product Code |
Product Name |
Package Size |
| P2983 |
ANTI-FLAG HS Clear M2 Coated Plate |
1 EA |
| P2983 |
ANTI-FLAG HS Clear M2 Coated Plate |
5 X 1 EA |
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Sample Data
Automated Capture and ELISA Analysis of a FLAG-fusion Protein
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| Figure 1. Standard curve generated from diluted FLAG-Bacterial Alkaline Phosphatase (BAP) standards. The automated Sciclone method was used to perform an ELISA. FLAG- BAP protein was serially diluted and then incubated at room temperature for 2 hours in the ANTI-FLAG M2 coated plate. For ELISA analysis, the BAP protein was first incubated with rabbit anti-alkaline phosphatase antibody for 1 hour at room temperature. Following four wash steps, the BAP protein was then incubated with the secondary antibody, a goat anti-rabbit IgG peroxidase conjugate, for 1 hour at room temperature. The peroxidase was detected by using a soluble TMB substrate for ELISA applications. The reaction was stopped using TMB stop solution. Absorbance was read on a Molecular Devices SpectraMax® Plus384 at 450 nm. |
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Cross Contamination Analysis
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| Figure 2. Cross contamination analysis using FLAG-BAP protein standard and blank wells. Cross contamination was evaluated in the automated ELISA method by inserting blanks between rows of FLAG-BAP protein standards. For ELISA analysis, the BAP protein was first incubated with rabbit anti-alkaline phosphatase antibody for 1 hour at room temperature. Following four wash steps, the BAP protein was then incubated with the secondary antibody, a goat anti-rabbit IgG peroxidase conjugate, for 1 hour at room temperature. The peroxidase was detected by using a soluble TMB substrate for ELISA applications. The reaction was stopped using TMB stop solution. Absorbance was read on a Molecular Devices SpectraMax® Plus384 at 450 nm. No cross contamination was detected in the blanks.
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