Preparation And Storage
Recommended Assay Procedures
Put all BD® AbSeq Reagents to be pooled into a Latch Rack for 500 µL Tubes (Thermo Fisher Scientific Cat. No. 4900). Arrange the tubes so that they can be easily uncapped and re-capped with an 8-Channel Screw Cap Tube Capper (Thermo Fisher Scientific Cat. No. 4105MAT) and the reagents aliquoted with a multi-channel pipette.
BD® AbSeq tubes should be centrifuged for ≥ 30 seconds at 400 × g to ensure removal of any content in the cap/tube threads prior to the first opening.
The P1H12 monoclonal antibody specifically binds to CD146. CD146 is a 118 kDa transmembrane glycoprotein also known as MCAM, MUC18, or Mel-CAM. CD146 is a member of the immunoglobulin superfamily strongly expressed by blood vessel endothelium and smooth muscle. CD146 is also expressed by angioblasts, mesenchymal stem cells, melanoma cells, intermediate trophoblasts and a subpopulation of activated T cells. The P1H12 monoclonal antibody has been reported to block endothelial cell adhesion that is observed very early in embryogenesis. It can be useful in the study of embryologic vasculogenesis. This antibody is suitable for immunohistochemical staining of acetone-fixed frozen tissue sections, immunoprecipitation and ELISA.
Application Notes
The antibody was conjugated to an oligonucleotide that contains an antibody clone-specific barcode (ABC) flanked by a poly-A tail on the 3' end and a PCR handle (PCR primer binding site) on the 5' end. The ABC for this antibody was designed to be used with other BD AbSeq oligonucleotides conjugated to other antibodies. All AbSeq ABC sequences were selected in silico to be unique from human and mouse genomes, have low predicted secondary structure, and have high Hamming distance within the BD AbSeq portfolio, to allow for sequencing error correction and unique mapping. The poly-A tail of the oligonucleotide allows the ABC to be captured by the BD Rhapsody™ system. The 5' PCR handle allows for efficient sequencing library generation for Illumina sequencing platforms.
NOTE: The BD Rhapsody Single-Cell Analysis System must be used with the BD Rhapsody Express Instrument.
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.