ReadiUse™ Preactivated PE-Cy5 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)	666

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 Maleimide

Reconstitute ReadiUse™ Preactivated PE-Cy5 Maleimide in 100 µL ddH₂O to 10 mg/mL. Note: Reconstituted ReadiUse™ Preactivated PE-Cy5 Maleimide are stable at 4 °C for one week, please kept it from light.
Add reduced antibody to pre-activated PE-Cy5 directly to at the ratio of 250-300 µg PE-Cy5 /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 conjugate could be further purified through size exclusion chromatography to get best performance. Note: The Antibody/PE-Cy5 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)	666

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated PE-Cy5 Tandem	565	666	1960000
ReadiUse™ Preactivated PE-Cy7 Maleimide	565	778	1960000

Images

Figure 1. Flow cytometry analysis of whole blood stained with PE-Cy5 anti-human CD4 *SK3* conjugate. The fluorescence signal was monitored using an Aurora spectral flow cytometer in the PE-Cy5 specific B8-A channel.

References

View all 50 references: Citation Explorer

Indicator of Inflammation and NETosis-Low-Density Granulocytes as a Biomarker of Autoimmune Hepatitis.
Authors: Domerecka, Weronika and Homa-Mlak, Iwona and Mlak, Radosław and Michalak, Agata and Wilińska, Agnieszka and Kowalska-Kępczyńska, Anna and Dreher, Piotr and Cichoż-Lach, Halina and Małecka-Massalska, Teresa
Journal: Journal of clinical medicine (2022)

Expression of CD163 and HLA-DR molecules on the monocytes in chronic lymphocytic leukemia patients.
Authors: Kowalska, Wioleta
Journal: Folia histochemica et cytobiologica (2020): 17-24

Anti-epithelial cell adhesion molecule RNA aptamer-conjugated liposomal doxorubicin as an efficient targeted therapy in mice bearing colon carcinoma tumor model.
Authors: Mashreghi, Mohammad and Zamani, Parvin and Karimi, Maryam and Mehrabian, Amin and Arabsalmani, Mahdieh and Zarqi, Javad and Moosavian, Seyedeh Alia and Jaafari, Mahmoud Reza
Journal: Biotechnology progress (2020): e3116

Image cytometry as an alternative to flow cytometry for the transplant histocompatibility crossmatch assay.
Authors: Ramon, Daniel S and Franks, Thomas and Jaramillo, Andrés and Paradis, Benjamin D and Chan, Leo Li-Ying
Journal: HLA (2019): 436-444

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

Association of Regulatory T Cells with Diabetes Type-1 and Its Renal and Vascular Complications Based on the Expression of Forkhead Box Protein P3 (FoxP3), Helios and Neurophilin-1.
Authors: Khamechian, Tahereh and Irandoust, Behnaz and Mohammadi, Hanieh and Nikoueinejad, Hassan and Akbari, Hossein
Journal: Iranian journal of allergy, asthma, and immunology (2018): 151-157

A Handy Liquid Metal Based Non-Invasive Electrophoretic Particle Microtrap.
Authors: Tian, Lu and Zhang, Lunjia and Gao, Meng and Deng, Zhongshan and Gui, Lin
Journal: Micromachines (2018)

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

CD4+ T cells and natural killer cells: Biomarkers for hepatic fibrosis in human immunodeficiency virus/hepatitis C virus-coinfected patients.
Authors: Laufer, Natalia and Ojeda, Diego and Polo, María Laura and Martinez, Ana and Pérez, Héctor and Turk, Gabriela and Cahn, Pedro and Zwirner, Norberto Walter and Quarleri, Jorge
Journal: World journal of hepatology (2017): 1073-1080

Multiplexed Lipid Bilayers on Silica Microspheres for Analytical Screening Applications.
Authors: Fernandez Oropeza, Nadiezda and Zurek, Nesia A and Galvan-De La Cruz, Mirella and Fabry-Wood, Aurora and Fetzer, Jennifer M and Graves, Steven W and Shreve, Andrew P
Journal: Analytical chemistry (2017): 6440-6447