CD304 (Neuropilin-1) Monoclonal Antibody (3DS304M), PerCP-eFluor™ 710, eBioscience™

Product Specific Information

Description: The monoclonal antibody 3DS304M recognizes mouse CD304 (Neuropilin-1), a type 1 transmembrane glycoprotein present on the surface of multiple cell types, including: regulatory T cells (Treg), NKT cells, DC, certain types of stem cells, neurons, endothelial cells, and some neoplastic cells. Neuropilin-1, in complex with plexins, serves as a co-receptor for type 3 semaphorins in growing neurons. It is also involved in the process of angiogenesis being a part of a functional receptor for VEGF in endothelial cells. In mice, Neuropilin-1 is expressed on thymus-derived natural Treg but not peripherally induced Treg. In addition, transient expression on recently activated non-regulatory T cells has been observed. Neuropilin-1 has been used as a marker of recent thymic emigrants in the mouse iNKT cell population. It has been shown that Neuropilin-1 forms a complex with TGF beta receptors, activating the latent form of TGF beta (LAP-TGF beta 1) and augmenting canonical Smad2/3 signaling.

This 3DS304M antibody will recognize formaldehyde-fixed as well as methanol-treated epitopes.

Applications Reported: This 3DS304M antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 3DS304M antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 0.06 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

Neuropilin 1 is a 923 amino acid containing membrane bound coreceptor to a tyrosine kinase receptor belonging to the neuropilin family with two CUB domains, two F5/8 type C domains and a MAM domain. Neuropilin 1 forms a heterodimer with NRP2 (Neuropilin 2) and is known to bind to plexin (PLXNB1) to form semaphorin receptors. Expressed mostly in the nervous system this receptor is implicated in a wide range of functions including in the development of the cardiovascular system, angiogenesis, repulsive axon guidance, formation of certain neuronal circuits, cell survival, control of neuronal migration and organogenesis outside the nervous system. This semaphorin receptor functions as receptors for both the semaphorins and vascular endothelial growth factor and is also a regulator of VEGF-induced angiogenesis. Neuropilin 1 may be involved in aggressive advanced prostate carcinoma and in the progression of breast cancer.