ReadiUse™ Preactivated APC Maleimide [Activated Allophycocyanin]

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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

Solvent

Water

Spectral properties

Correction Factor (280 nm)	0.195
Extinction coefficient (cm ^-1 M ^-1)	730000
Excitation (nm)	651
Emission (nm)	660

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
UNSPSC	12171501

Overview SDS Protocol

Example protocol

SAMPLE EXPERIMENTAL PROTOCOL

Reduction of Antibody

Prepare a fresh solution of 1.0 M DTT (15.4 mg/100 µL) in distilled water. Antibody reduction can be carried out in different buffers, such as MES, phosphate, or TRIS (pH 6 to 8). For best results, antibody solutions should be >2 mg/mL. If less than 2 mg/mL, the antibody should be concentrated.
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 temperature for 30 minutes without additional mixing to minimize the reoxidation of cysteines.
Purify the reduced antibody using a desalting column (Cat No. 60500) pre-equilibrated with 50 mM MES buffer (pH 6.0-6.5) and 2 mM EDTA.
Measure the antibody concentration using a Nanodrop. (Con. (mg/mL) = A280nm/1.4).
Note Reduced antibodies are typically unstable. It is best to run the conjugation reaction immediately following purification.

Conjugate with ReadiUse™ Preactivated APC Maleimide

Reconstitute ReadiUse™ Preactivated APC Maleimide in ddH2O to make a 10 mg/mL solution. For Cat No. 2567 (1 mg) reconstitute in 100 µL of ddH2O. If using Cat No. 2568 (5 mg), reconstitute in 500 µL of ddH2O. Reconstituted ReadiUse™ Preactivated APC Maleimide solutions are stable at 4 °C for up to one month, protected from light.
Add the reduced antibody directly to the solution of ReadiUse™ Preactivated APC at a ratio of 130 µg APC/100 µg of reduced antibody.
Rotate the mixture for 60 to 120 minutes at room temperature.
Block any free sulfhydryl groups on the antibody. Prepare a fresh solution of 10 mg/mL N-Ethylmaleimide (NEM) in DMSO and add 3.4 µL per mg of antibody. Rotate for 20 minutes at room temperature.

Conjugate Purification

Purify the antibody/APC conjugate solution using size exclusion chromatography.
Note Typically the antibody/APC conjugate can be stored at 2-8 °C, protected from light, for up to 6 months. The best storage conditions for any particular conjugate must be determined by experimentation and depends on the stability of the antibody.

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Correction Factor (280 nm)	0.195
Extinction coefficient (cm ^-1 M ^-1)	730000
Excitation (nm)	651
Emission (nm)	660

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)	Correction Factor (280 nm)
ReadiUse™ Preactivated PE Maleimide [Activated R-Phycoerythrin]	565	574	1960000	0.175
ReadiUse™ Preactivated PerCP Maleimide	477	678	406000	0.22

Images

Figure 1. Flow cytometry analysis of whole blood cells stained with CD8 (SK1)-APC conjugate prepared with ReadiUse™ Preactivated APC Maleimide (Cat No. 2567). The fluorescence signal was monitored using an Aurora spectral flow cytometer in the R1-A channel

References

View all 13 references: Citation Explorer

Magnetic particles encoding a suspension probe for ultra-sensitive and quantitative determination of atrazine.
Authors: Zhao, Zhigao and Dou, Xiaowen and Luo, Jiaoyang and Jin, Meiqi and Qin, Jiaan and Wang, Changjian and Yang, Shihai and Yang, Meihua
Journal: Journal of pharmaceutical and biomedical analysis (2021): 113868

Non-oxidative band-3 clustering agents cause the externalization of phosphatidylserine on erythrocyte surfaces by a calcium-independent mechanism.
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Leukocyte-Derived Extracellular Vesicles in Blood with and without EpCAM Enrichment.
Authors: Nanou, Afroditi and Zeune, Leonie L and Terstappen, Leon W M M
Journal: Cells (2019)

Exogenous IL-9 Ameliorates Experimental Autoimmune Myasthenia Gravis Symptoms in Rats.
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Journal: Immunological investigations (2018): 712-724

Non-invasive dual fluorescence in vivo imaging for detection of macrophage infiltration and matrix metalloproteinase (MMP) activity in inflammatory arthritic joints.
Authors: Cho, Hongsik and Bhatti, Fazal-Ur-Rehman and Yoon, Tae Won and Hasty, Karen A and Stuart, John M and Yi, Ae-Kyung
Journal: Biomedical optics express (2016): 1842-52

Monitoring genotoxicity in patients receiving chemotherapy for cancer: application of the PIG-A assay.
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Journal: Mutation research. Genetic toxicology and environmental mutagenesis (2016): 20-6

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Journal: Scientific reports (2016): 25141

Endothelial activation and apoptosis mediated by neutrophil-dependent interleukin 6 trans-signalling: a novel target for systemic sclerosis?
Authors: Barnes, Theresa C and Spiller, David G and Anderson, Marina E and Edwards, Steven W and Moots, Robert J
Journal: Annals of the rheumatic diseases (2011): 366-72

A new procedure for the quantitative assessment of p-glycoprotein efflux pump associated with human T lymphocytes.
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Journal: Ethnicity & disease (2008): S2-75-80

Dextramers: new generation of fluorescent MHC class I/peptide multimers for visualization of antigen-specific CD8+ T cells.
Authors: Batard, Pascal and Peterson, Daniel A and Devêvre, Estelle and Guillaume, Philippe and Cerottini, Jean-Charles and Rimoldi, Donata and Speiser, Daniel E and Winther, Lars and Romero, Pedro
Journal: Journal of immunological methods (2006): 136-48