ReadiUse™ Preactivated APC-AF700 Maleimide

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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

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 Antibody

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 APC-AF700 Maleimide

Reconstitute ReadiUse™ Preactivated APC-AF700 Maleimide in 100 µL ddH₂O to 10 mg/mL. Note: Reconstituted ReadiUse™ Preactivated APC-AF700 Maleimide are stable at 4 °C for one week, please kept it from light.
Add reduced antibody to pre-activated APC-AF700 directly to at the ratio of 130 µg APC-AF700 /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/APC-AF700 conjugate could be further purified through size exclusion chromatography to get best performance. Note: The Antibody/APC-AF700 conjugate solution is recommended to be stored at 2~8 °C and kept from light.

Product Family

Name	Excitation (nm)	Emission (nm)	Extinction coefficient (cm ^-1 M ^-1)
ReadiUse™ Preactivated APC-Cy7 Maleimide	651	779	700000

References

View all 8 references: Citation Explorer

Bivalent EGFR-Targeting DARPin-MMAE Conjugates.
Authors: Karsten, Lennard and Janson, Nils and Le Joncour, Vadim and Alam, Sarfaraz and Müller, Benjamin and Tanjore Ramanathan, Jayendrakishore and Laakkonen, Pirjo and Sewald, Norbert and Müller, Kristian M
Journal: International journal of molecular sciences (2022)

Detecting and predicting neutralization of alemtuzumab responses in MS.
Authors: Saxena, Gauri and Moore, James M and Jones, Meleri and Pryce, Gareth and Ali, Liaqat and Leisegang, Georgia R and Vijay, Vivek and Loveless, Samantha and Robertson, Neil P and Schmierer, Klaus and Giovannoni, Gavin and Gnananpavan, Sharmilee and Baker, David and Tallantyre, Emma C and Kang, Angray S
Journal: Neurology(R) neuroimmunology & neuroinflammation (2020)

Activity-Based Protein Profiling of Ammonia Monooxygenase in Nitrosomonas europaea.
Authors: Bennett, Kristen and Sadler, Natalie C and Wright, Aaron T and Yeager, Chris and Hyman, Michael R
Journal: Applied and environmental microbiology (2016): 2270-2279

Pharmacokinetic Profile and Acute Toxicological Properties of a Novel Radiosensitizer Cytosine-Phosphate-Guanosine Oligodeoxynucleotide 107 in Mice Following Intravenous and Orthotopic Administration.
Authors: Cen, Yanyan and Li, Xiaoli and Yin, Zhiwei and Yan, Zifei and Liu, Dan and Peng, Wei and Pan, Feng and Zhou, Hong
Journal: Nucleic acid therapeutics (2015): 254-65

Membrane labeling of coral gastrodermal cells by biotinylation: the proteomic identification of surface proteins involving cnidaria-dinoflagellate endosymbiosis.
Authors: Li, Hsing-Hui and Huang, Zi-Yu and Ye, Shih-Png and Lu, Chi-Yu and Cheng, Pai-Chiao and Chen, Shu-Hwa and Chen, Chii-Shiarng
Journal: PloS one (2014): e85119

Selective 351 nm photodissociation of cysteine-containing peptides for discrimination of antigen-binding regions of IgG fragments in bottom-up liquid chromatography-tandem mass spectrometry workflows.
Authors: Cotham, Victoria C and Wine, Yariv and Brodbelt, Jennifer S
Journal: Analytical chemistry (2013): 5577-85

Changes of cell-surface thiols and intracellular signaling in human monocytic cell line THP-1 treated with diphenylcyclopropenone.
Authors: Hirota, Morihiko and Motoyama, Akira and Suzuki, Mie and Yanagi, Masashi and Kitagaki, Masato and Kouzuki, Hirokazu and Hagino, Shigenobu and Itagaki, Hiroshi and Sasa, Hitoshi and Kagatani, Saori and Aiba, Setsuya
Journal: The Journal of toxicological sciences (2010): 871-9

Tandem dye acceptor used to enhance upconversion fluorescence resonance energy transfer in homogeneous assays.
Authors: Rantanen, Terhi and Päkkilä, Henna and Jämsen, Laura and Kuningas, Katri and Ukonaho, Telle and Lövgren, Timo and Soukka, Tero
Journal: Analytical chemistry (2007): 6312-8