ReadiUse™ Preactivated PE-Cy7 Maleimide

Ordering information

Price
Catalog Number
Unit Size
Quantity

Add to cart

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)	778

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-Cy7 Maleimide

Reconstitute ReadiUse™ Preactivated PE-Cy7 Maleimide in 100 µL ddH₂O to 10 mg/mL. Note: Reconstituted ReadiUse™ Preactivated PE-Cy7 Maleimide are stable at 4 °C for one week, please kept it from light.
Add reduced antibody to pre-activated PE-Cy7 directly to at the ratio of 250-300 µg PE-Cy7 /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-Cy7 conjugate could be further purified through size exclusion chromatography to get best performance. Note: The Antibody/PE-Cy7 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)	778

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated PE-Cy7 Tandem	565	778	1960000
ReadiUse™ Preactivated PE-Cy5 Maleimide	565	666	1960000
ReadiUse™ Preactivated APC-Cy7 Maleimide	651	779	700000
ReadiUse™ Preactivated PerCP-Cy7 Maleimide	757	783	350000

Images

Figure 1. Flow cytometry analysis of whole blood stained with PE-Cy7 anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora spectral flow cytometer in the PE-Cy7 specific B13-A channel.

References

View all 37 references: Citation Explorer

Recombinant thrombomodulin attenuates hyper-inflammation and glycocalyx damage in a murine model of Streptococcus pneumoniae-induced sepsis.
Authors: Watanabe, Eizo and Akamatsu, Toshinobu and Ohmori, Masaaki and Kato, Mayu and Takeuchi, Noriko and Ishiwada, Naruhiko and Nishimura, Rintaro and Hishiki, Haruka and Fujimura, Lisa and Ito, Chizuru and Hatano, Masahiko
Journal: Cytokine (2022): 155723

Cordyceps militaris extract induces apoptosis and pyroptosis via caspase-3/PARP/GSDME pathways in A549 cell line.
Authors: Hu, Zixuan and Lai, Yijing and Ma, Chaoya and Zuo, Lina and Xiao, Guanlin and Gao, Haili and Xie, Biyuan and Huang, Xuejun and Gan, Haining and Huang, Dane and Yao, Nan and Feng, Baoguo and Ru, JieXia and Chen, Yuxing and Cai, Dake
Journal: Food science & nutrition (2022): 21-38

Development and validation of a sensitive flow cytometric method for determining CECs in RBC products.
Authors: Li, Wenhui and Acker, Jason P
Journal: Clinica chimica acta; international journal of clinical chemistry (2022): 119-125

Routine flow cytometry approach for the evaluation of solid tumor neoplasms and immune cells in minimally invasive samples.
Authors: Quirós-Caso, Covadonga and Arias Fernández, Tamara and Fonseca-Mourelle, Ariana and Torres, Héctor and Fernández, Luis and Moreno-Rodríguez, Maria and Ariza-Prota, Miguel Ángel and López-González, Francisco Julián and Carvajal-Álvarez, Miguel and Alonso-Álvarez, Sara and Moro-García, Marco Antonio and Colado, Enrique
Journal: Cytometry. Part B, Clinical cytometry (2022)

The ISCCA flow protocol for the monitoring of anti-CD20 therapies in autoimmune disorders.
Authors: Gatti, Arianna and Buccisano, Francesco and Scupoli, Maria T and Brando, Bruno
Journal: Cytometry. Part B, Clinical cytometry (2021): 194-205

Fluorescent energy transfer causing misleading signal in multicolor flow cytometry.
Authors: Khenine, Hana and Waeckel, Louis and Seghrouchni, Fouad and Berger, Anne-Emmanuelle and Lambert, Claude
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2021)

Probing Surface Membrane Receptors Using Engineered Bacteriophage Bioconjugates.
Authors: Lomakin, Yakov A and Kaminskaya, Alena N and Stepanov, Alexey V and Shmidt, Anna A and Gabibov, Alexander G and Belogurov, Alexey A
Journal: Bioconjugate chemistry (2019): 1500-1506

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

Reduced CD4 T Lymphocytes in Lymph Nodes of the Mouse Model of Autism Induced by Valproic Acid.
Authors: Baronio, Diego and Bauer-Negrini, Guilherme and Castro, Kamila and Della-Flora Nunes, Gustavo and Riesgo, Rudimar and Mendes-da-Cruz, Daniella Arêas and Savino, Wilson and Gottfried, Carmem and Bambini-Junior, Victorio
Journal: Neuroimmunomodulation (2018): 280-284

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