ReadiUse™ Preactivated PE-Cy5.5 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)	671

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-Cy5.5 Maleimide

Reconstitute ReadiUse™ Preactivated PE-Cy5.5 Maleimide in 100 µL ddH₂O to 10 mg/mL. Note: Reconstituted ReadiUse™ Preactivated PE-Cy5.5 Maleimide are stable at 4 °C for one week, please kept it from light.
Add reduced antibody to pre-activated PE-Cy5.5 directly to at the ratio of 250-300 µg PE-Cy5.5 /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-Cy5.5 conjugate could be further purified through size exclusion chromatography to get best performance. Note: The Antibody/PE-Cy5.5 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)	671

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated PE-Cy5.5 Tandem	565	671	1960000
ReadiUse™ Preactivated APC-Cy5.5 Maleimide	651	700	700000
ReadiUse™ Preactivated PerCP-Cy5.5 Maleimide	482	695	350000

References

View all 5 references: Citation Explorer

Initial flow cytometric evaluation of the Clearllab lymphoid screen.
Authors: Hedley, B D and Cheng, G and Luider, J and Kern, W and Lozanski, G and Chin-Yee, I and Lowes, L E and Keeney, M and Careaga, D and Magari, R and Tejidor, L
Journal: Cytometry. Part B, Clinical cytometry (2018): 707-713

A ten-color tube with dried antibody reagents for the screening of hematological malignancies.
Authors: Correia, R P and Rajab, A and Bento, L C and Alexandre, A M and Vaz, A C and Schimidell, D and Pedro, E C and Perin, F S and Nozawa, S T and Barroso, R S and Bacal, N S
Journal: International journal of laboratory hematology (2018): 136-143

Simultaneous flow cytometric measurement of antigen attachment to phagocytes and phagocytosis.
Authors: Laopajon, Witida and Takheaw, Nuchjira and Kasinrerk, Watchara and Pata, Supansa
Journal: Journal of immunoassay & immunochemistry (2016): 527-39

PE-Cy5.5 conjugates bind to the cells expressing mouse DEC205/CD205.
Authors: Park, Chae Gyu and Rodriguez, Anthony and Steinman, Ralph M
Journal: Journal of immunological methods (2012): 184-90

Polychromatic (eight-color) slide-based cytometry for the phenotyping of leukocyte, NK, and NKT subsets.
Authors: Mittag, Anja and Lenz, Dominik and Gerstner, Andreas O H and Sack, Ulrich and Steinbrecher, Michael and Koksch, Mario and Raffael, Alexander and Bocsi, Jozsef and Tárnok, Attila
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2005): 103-15