ReadiUse™ Preactivated PE-Texas Red Maleimide

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Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Physical properties

Molecular weight	N/A
Solvent	Water

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	613

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
Storage	Refrigerated (2-8 °C); Minimize light exposure

Overview SDS Protocol

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

Open in Advanced Spectrum Viewer

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	1960000
Excitation (nm)	565
Emission (nm)	613

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated PE-Texas Red Tandem	565	613	1960000

Images

Figure 1. Flow cytometry analysis of whole blood stained with PE-Texas Red anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora spectral flow cytometer in the PE-Texas Red specific B6-A channel.

References

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Flow cytometry can diagnose classical hodgkin lymphoma in lymph nodes with high sensitivity and specificity.
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Journal: American journal of clinical pathology (2009): 322-32

Optimization of three- and four-color multiparameter DNA analysis in lymphoma specimens.
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Multiparameter cytokine-specific affinity matrix assay for the determination of frequencies and phenotype of antigen-reactive T cells.
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Journal: Journal of immunological methods (2002): 37-42

Conjugation of fluorochromes to monoclonal antibodies.
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Journal: Current protocols in cytometry (2001): Unit 4.2

A strategy for multiple immunophenotyping by image cytometry: model studies using latex microbeads labeled with seven streptavidin-bound fluorochromes.
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Journal: Cytometry (1996): 214-25

CUBIC: a three-dimensional colored projection of Consort 30 generated trivariate flow cytometric data.
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Journal: Cytometry (1991): 570-8

Concomitant delineation of surface Ig, B-cell differentiation antigens, and HLADR on lymphoid proliferations using three-color immunocytometry.
Authors: Segal, G H and Edinger, M G and Owen, M and McNealis, M and Lopez, P and Perkins, A and Linden, M D and Fishleder, A J and Stoler, M H and Tubbs, R R
Journal: Cytometry (1991): 350-9