Stop Solution for TMB Substrates Reagent | STOPT reagent
Stop Solution for TMB Substrates (Liquid)
Synonyms
Stop Solution for TMB Substrates; N/A; Stop Solution for TMB Substrates (Liquid); Stop Solution for TMB Substrates (STOPT); STOPT reagent
Form/Format
Clear, colorless liquid
Instructions
1. Run ELISA according to the specific protocol through the conjugate incubation step.
2. Wash the wells three or four times with 1X ELISA Wash Buffer to remove any residual HRP-conjugate.
3. Add TMB substrate to each well of the plate. For example, use 100 uL/well; protect from light.
4. Incubate the substrate 10-60 minutes. Monitor the color intensity.
5. Stop the reaction by adding an equal volume of STOPT to the substrate in every well of the plate, e.g., if each well contains 100 uL substrate, add 100 uL/well STOPT.
6. Read the plate at 450 nm within 1 hour.
2. Wash the wells three or four times with 1X ELISA Wash Buffer to remove any residual HRP-conjugate.
3. Add TMB substrate to each well of the plate. For example, use 100 uL/well; protect from light.
4. Incubate the substrate 10-60 minutes. Monitor the color intensity.
5. Stop the reaction by adding an equal volume of STOPT to the substrate in every well of the plate, e.g., if each well contains 100 uL substrate, add 100 uL/well STOPT.
6. Read the plate at 450 nm within 1 hour.
pH
<2; 1X ready to use; Read absorbance at 450 nm
Safty and Usage
Contains Maleic acid at 3-7%
Preparation and Storage
Store at 25 degree C
Refrigerated temperatures will not harm the reagent.
Refrigerated temperatures will not harm the reagent.
Related Product Information for STOPT reagent
Description: Stop and stabilize TMB microwell substrate reactions.
Stop Solution for TMB Substrates (STOPT) is a proprietary acidic formulation used to stop TMB microwell substrate reactions. It is suitable for all endpoint ELISAs using a tetramethylbenzidine (TMB) substrate reaction for color development.
TMB substrates are oxidized by peroxidase enzymes to yield a soluble blue-green reaction product. In endpoint assays, the reaction can be stopped by adding equal volumes of Stop Solution for TMB Substrates. Addition of STOPT changes the chromagen color from blue-green to yellow, where it can be read at 450 nm, and concurrently stabilizes the yellow TMB product for one hour. Stopping the reaction will increase the sample absorbance value up to 3-fold. To avoid overdeveloping the TMB substrate reaction, the blue-green reaction product should be periodically monitored on an ELISA plate reader using 620-650 nm absorbance filter settings. When OD values reach approximately 0.7 units, the reaction should be stopped with STOPT.
For best results, the absorbance should be monitored and read before values exceed 2.5 OD units. If the yellow reaction product yields OD values above 2.5 units, it is recommended to dilute the antibodies/conjugates or shorten the incubation period.
Stop Solution for TMB Substrates is ready to use at 1X. Stop the reaction by adding an equal volume of STOPT to the substrate in every well of the plate.
Stop Solution for TMB Substrates (STOPT) is a proprietary acidic formulation used to stop TMB microwell substrate reactions. It is suitable for all endpoint ELISAs using a tetramethylbenzidine (TMB) substrate reaction for color development.
TMB substrates are oxidized by peroxidase enzymes to yield a soluble blue-green reaction product. In endpoint assays, the reaction can be stopped by adding equal volumes of Stop Solution for TMB Substrates. Addition of STOPT changes the chromagen color from blue-green to yellow, where it can be read at 450 nm, and concurrently stabilizes the yellow TMB product for one hour. Stopping the reaction will increase the sample absorbance value up to 3-fold. To avoid overdeveloping the TMB substrate reaction, the blue-green reaction product should be periodically monitored on an ELISA plate reader using 620-650 nm absorbance filter settings. When OD values reach approximately 0.7 units, the reaction should be stopped with STOPT.
For best results, the absorbance should be monitored and read before values exceed 2.5 OD units. If the yellow reaction product yields OD values above 2.5 units, it is recommended to dilute the antibodies/conjugates or shorten the incubation period.
Stop Solution for TMB Substrates is ready to use at 1X. Stop the reaction by adding an equal volume of STOPT to the substrate in every well of the plate.
Product Categories/Family for STOPT reagent
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Product Notes
The STOPT (Catalog #AAA52746) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "Stop Solution for TMB Substrates, Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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