Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis and the oligomer sequence is confirmed by sequencing. TotalSeq?-A antibodies are compatible with 10x Genomics Single Cell Gene Expression Solutions.

To maximize performance, it is strongly recommended that the reagent be titrated for each application, and that you centrifuge the antibody dilution before adding to the cells at 14,000xg at 2 - 8°C for 10 minutes. Carefully pipette out the liquid avoiding the bottom of the tube and add to the cell suspension. For Proteogenomics analysis, the suggested starting amount of this reagent for titration is ≤ 1.0 ?g per million cells in 100 ?L volume. Refer to the corresponding TotalSeq? protocol for specific staining instructions.

This antibody is effective in immunoblotting (WB), immunohistochemistry (IHC), immunofluorescence (IF), immunoprecipitation (IP) and immunoelectron microscopy (IEM). This antibody has been used successfully with frozen sections.

*Predicted MW = 62 kD

MAb414 is a reliable general purpose monoclonal antibody which recognizes a related family of NPC proteins. This antibody is ideal for studying the morphology and composition of the nucleus and nuclear envelope. It is also useful in studying changes in the nuclear structure during mitosis and meiosis.

Hashtag reagents require different primers than regular TotalSeq? antibodies. Please refer to the protocol for details.

TotalSeq? Hashtag reagents can be used for multiplexing different samples; optimization of microfluidics instruments to load a high number of cells, and discriminate droplets containing more than one cell; performing TotalSeq? antibody titrations, etc. A compatible single cell device (e.g. 10x Genomics Chromium System and Reagents) and sequencer (e.g. Illumina analyzers) are required. Please contact technical support for more information, or visit biolegend.com/totalseq.

The barcode flanking sequences are GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT (PCR handle), and BAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA*A*A (capture sequence). B represents either C, G, or T, and * indicates a phosphorothioated bond, to prevent nuclease degradation.

View more applications data for this product in our Scientific Poster Library.

1. Zheng X, et al. 2012. J. Biol. Chem. 287:38254. (IF) PubMed
2. Kimura T, et al. 2003. Mol Cell Biol. 23:1304. (IHC) PubMed
3. Lopez-Soler RI, et al. 2001. J Cell Biol. 154:61. (IF, WB) PubMed
4. Aris JP, Blobel G. 1989. J Cell Biol. 108:2059. (WB, IP, IF, EM) PubMed
5. Davis LI, Blobel G. 1987. PNAS USA. 84:7552. (IP, IF) PubMed
6. Edens LJ, Levy DL. 2014. J. Cell. Biol. 206:473.
7. Davis LI, Blobel G. 1986. Cell. 45:699.?
8. Blobel G. 1985. PNAS USA 82:8527.

1. Yoshimura S, et al. 2013. J. Cell Sci. 126:3141-50.