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ReadiUse™ Preactivated PE-Texas Red Maleimide
PE-Texas Red is a popular tandem color used in flow cytometry. Its primary absorption peak is at 565 nm with emission peak at 600 nm. AAT Bioquest offers this preactivated PE-Texas Red maleimide to facilitate the PE-Texas Red tandem conjugations to antibodies and other proteins such as streptavidin and other secondary reagents. It is prepared from the commonly used crosslinker SMCC. Our preactivated PE-Texas Red maleimide is ready to conjugate the reduced antibodies and other thiol-containing biomolecules.
Example protocol
SAMPLE EXPERIMENTAL PROTOCOL
Reduction of IgG
- Prepare a fresh solution of 1.0 M DTT (15.4 mg/100 µL) in distilled water. Antibody solutions should be at 2 mg/mL or higher for best results. The reduction can be carried out in different buffers for example: MES, phosphate, and TRIS buffers (pH range 6 to 8). The antibody should be concentrated if less than 2 mg/mL.
- Add 2 µL of 1.0 M DTT stock per 100 µL of antibody solution and mix well. Let the antibody solution stand at room temp for 30 minutes without additional mixing (to minimize reoxidation of cysteines to cystines).
- Purify the reduced antibody over a desalting column pre-equilibrated with 50 mM MES Buffer (pH=6.0-6.5) with 2 mM EDTA. (Desalting column: https://www.aatbio.com/products/readiuse-bio-gel-p-6-spin-column?unit=60500)
- Measure the Antibody concentration with Nanodrop. (Con. (mg/ml)= A280nm/1.4) Note: The reduced antibody is not stable; the conjugation reaction needs to be carried out the conjugation as soon as possible after purification.
Conjugate with ReadiUse™ Preactivated PE-Texas Red Maleimide
- Reconstitute ReadiUse™ Preactivated PE-Texas Red Maleimide in 100 µL ddH2O to 10 mg/mL. Note: Reconstituted ReadiUse™ Preactivated PE-Texas Red Maleimide are stable at 4 °C for one week, please kept it from light.
- Add reduced antibody to pre-activated PE-Texas Red directly to at the ratio of 250-300 µg PE-Texas Red /100 µg reduced antibody.
- Rotate the mixture for 60-120 mins at room temperature.
- After 60 minutes, block the free sulfhydryls on the antibody.
- Prepare a fresh solution of 10 mg/mL NEM in DMSO; add 3.4 µL per mg of antibody and rotate for 20 minutes at room temperature.
Purification
- Antibody/PE-Texas Red conjugate could be further purified through size exclusion chromatography to get best performance. Note: The Antibody/PE-Texas Red conjugate solution is recommended to be stored at 2~8 °C and kept from light.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) |
| ReadiUse™ Preactivated PE-Texas Red Tandem | 565 | 613 | 1960000 |
References
View all 8 references: Citation Explorer
Orange juice and its major polyphenol hesperidin consumption do not induce immunomodulation in healthy well-nourished humans.
Authors: Perche, Olivier and Vergnaud-Gauduchon, Juliette and Morand, Christine and Dubray, Claude and Mazur, Andrzej and Vasson, Marie-Paule
Journal: Clinical nutrition (Edinburgh, Scotland) (2014): 130-5
Authors: Perche, Olivier and Vergnaud-Gauduchon, Juliette and Morand, Christine and Dubray, Claude and Mazur, Andrzej and Vasson, Marie-Paule
Journal: Clinical nutrition (Edinburgh, Scotland) (2014): 130-5
Flow cytometry can diagnose classical hodgkin lymphoma in lymph nodes with high sensitivity and specificity.
Authors: Fromm, Jonathan R and Thomas, Anju and Wood, Brent L
Journal: American journal of clinical pathology (2009): 322-32
Authors: Fromm, Jonathan R and Thomas, Anju and Wood, Brent L
Journal: American journal of clinical pathology (2009): 322-32
Optimization of three- and four-color multiparameter DNA analysis in lymphoma specimens.
Authors: Plander, M and Brockhoff, G and Barlage, S and Schwarz, S and Rothe, G and Knuechel, R
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2003): 66-74
Authors: Plander, M and Brockhoff, G and Barlage, S and Schwarz, S and Rothe, G and Knuechel, R
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2003): 66-74
Multiparameter cytokine-specific affinity matrix assay for the determination of frequencies and phenotype of antigen-reactive T cells.
Authors: Mathioudakis, George and Coder, David and Fefer, Alexander
Journal: Journal of immunological methods (2002): 37-42
Authors: Mathioudakis, George and Coder, David and Fefer, Alexander
Journal: Journal of immunological methods (2002): 37-42
Conjugation of fluorochromes to monoclonal antibodies.
Authors: Holmes, K L and Lantz, L M and Russ, W
Journal: Current protocols in cytometry (2001): Unit 4.2
Authors: Holmes, K L and Lantz, L M and Russ, W
Journal: Current protocols in cytometry (2001): Unit 4.2
Page updated on November 16, 2024
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