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Neuropilin-1 Overexpression Lysate

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-13799

Novus Biologicals, part of Bio-Techne
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NBL1-13799

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Neuropilin-1 Overexpression Lysate

Neuropilin-1 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC203508

Accession#: NM_001024628

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

TMW

71.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for Neuropilin-1 Overexpression Lysate

Western Blot: Neuropilin-1 Overexpression Lysate [NBL1-13799]

Western Blot: Neuropilin-1 Overexpression Lysate [NBL1-13799]

Western Blot: Neuropilin-1 Overexpression Lysate [NBL1-13799] - Neuropilin 1 Overexpression Lysate (Adult Normal) [NBL1-13799]. Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Neuropilin 1. Theoretical molecular weight 103 kDa. Observed molecular weight ~70 kDa.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Neuropilin-1

Neuropilin-1 (also known as Npn-1, NRP1, neuropilin, or CD304/BDCA4) is a 923 amino acid (aa) type I transmembrane glycoprotein (theoretical molecular weight 130-140 kDa) located on human chromosome 10p11.22 that encodes multiple intact and soluble isoforms. The 623 aa extracellular domain (ECD) of this human cell surface receptor includes two CUB (complement-binding) domains, two F5/8 domains with homology to coagulation factors V and VIII, and a MAM (meprin) domain. The ECD of human Neuropilin-1 shares 92-95% aa sequence identity with mouse, rat, bovine, and canine Npn-1. The MAM domains of neuropilins are involved in the formation of homo- and hetero-oligomers in the absence of ligands. Neuropilin-1 expression is found in neuronal, endothelial and tumor cells as well as epithelial cells such as in the respiratory, gastrointestinal, lower urological tracts, thyroid, parathyroid and thymus gland (1,2).
Initially found to play an essential role in axon growth and guidance, NRP1 serves as a multifunctional co-receptor for class III semaphorins and heparin-binding members of the VEGF family, participating in regulation of angiogenesis, neuronal development, cell survival, migration and tumor-invasion. Neuropilin-1 has been implicated in viral infections and T-cell activation and is also described as a marker of regulatory T (Treg) cells (3). In COVID-19 infections, studies have shown NRP1 participates in viral infection via NRP1 neutralizing antibodies and binds the SARS-CoV-2 Spike protein, suggesting NRP1 may contribute to viral transport and viral internalization by endocytosis (4).

References

1. Wild JRL, Staton CA, Chapple K, Corfe BM. (2012) Neuropilins: Expression and Roles in the Epithelium. Int J Exp Pathol. 93(2):81-103. PMID: 22414290

2. Bielenberg DR, Pettaway CA, Takashima S, Klagsbrun M. (2006) Neuropilins in Neoplasms: Expression, Regulation, and Function. Exp Cell Res. 312(5):584-93. PMID: 16445911

3. Tordjman R, Lepelletier Y, Lemarchandel V, Cambot M, Gaulard P, Hermine O, Romeo P-H. (2002). A Neuronal Receptor, neuropilin-1, Is Essential for the Initiation of the Primary Immune Response. Nat Immunol. 3(5):477-82. PMID: 11953749

4. Coronavirus research updates: Test frequency matters more than test sensitivity for stopping outbreaks. 2002 June 26. Nature.com

Alternate Names

BDCA-4, CD304, Neuropilin1, NRP1

Gene Symbol

NRP1

Additional Neuropilin-1 Products

Product Documents for Neuropilin-1 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Neuropilin-1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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