Recombinant Human NT-4 GMP Protein, CF Summary
Manufactured and tested under current Good Manufacturing Practice (GMP) guidelines.
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
|Formulation||Lyophilized from a 0.2 μm filtered solution in Acetonitrile and TFA.|
|Reconstitution||Reconstitute at 100 μg/mL in PBS.|
|Shipping||The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.|
|Stability & Storage:||Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
GMP-grade Recombinant Human NT-4 (Catalog # 268-GMP) stimulates proliferation of the BaF mouse pro-B cell line transfected with TrkB. The ED50 for this effect is
1 μg/lane of Recombinant Human
GMP-grade NT-4 (Catalog # 268-GMP) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a band at 15 kDa.
Neurotrophin-4 (NT-4), also known as NT-5, is a member of the NGF family of neuronal and epithelial growth factors. Neurotrophins have six conserved cysteine residues that are involved in the formation of three disulfide bonds (1-3). The human NT-4 cDNA encodes a 210 amino acid (aa) precursor that includes a 24 aa signal sequence, a 56 aa propeptide, and a 130 aa mature protein (4, 5). NT-4 is synthesized as a 28 kDa prepropeptide that is proteolytically processed to generate the mature protein. Mature human NT-4 shares 48-52% aa sequence identity with human beta-NGF, BDNF, and NT-3. It shares 91% and 95% aa sequence identity with mouse and rat NT-4/5, respectively. The mature protein is secreted as a homodimer and can also form heterodimers with BDNF or NT-3 (6). NT-4 binds and induces receptor dimerization and activation of TrkB (4, 7). NT-4 promotes the development and survival of selected peripheral and CNS neurons (8-10). BDNF, which also activates TrkB, overlaps with many but not all NT-4 functions, a distinction that is likely due to differences in expression patterns (8-10). NT-4 induced TrkB signaling augments NMDA receptor activity and increases neuronal sensitivity to excitotoxic cell death (11). It also promotes the proliferation of keratinocytes and accelerates hair follicle regression during the follicular cycle (12, 13). NT-4 is secreted by activated T cells and granulocytes at sites of inflammation where it contributes to tissue regeneration (14-16).
- Lessmann, V. et al. (2003) Prog. Neurobiol. 69:341.
- Tabakman, R. et al. (2004) Prog. Brain Res. 146:387.
- Botchkarev, V.A. et al. (2004) Prog. Brain Res. 146:493.
- Ip, N.Y. et al. (1992) Proc. Natl. Acad. Sci. 89:3060.
- Berkemeier, L.R. et al. (1991) Neuron 7:857.
- Radziejewski, C. and R.C. Robinson (1993) Biochemistry 32:13350.
- Vesa, J. et al. (2000) J. Biol. Chem. 275:24414.
- Davies, A.M. et al. (1993) J. Neurosci. 13:4961.
- Stucky, C.L. et al. (1998) J. Neurosci. 18:7040.
- Fan, G. et al. (2000) Nat. Neurosci. 3:350.
- Choi, S.Y. et al. (2004) J. Neurochem. 88:708.
- Botchkarev, V.A. et al. (1999) Lab. Invest. 79:557.
- Botchkarev, V.A. et al. (1999) FASEB J. 13:395.
- Laurenzi, M.A. et al. (1998) J. Leukoc. Biol. 64:228.
- Moalem, G. et al. (2000) J. Autoimmun. 15:331.
- Nassenstein, C. et al. (2003) J. Exp. Med. 198:455.
Manufacturing SpecificationsGMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
R&D Systems' quality focus includes:
- Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
- Documented processes and QA control of documentation and process changes
- Personnel training programs
- Raw material testing and vendor qualification/monitoring
- Fully validated equipment, processes and test methods
- Equipment calibration schedules using a computerized calibration program
- Facility maintenance, safety programs and pest control
- Material review process for variances
- Monitoring of stability over product shelf-life
- N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
- Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial bioburden testing (using broth culture, Sabourand's dextrose and blood agar plates with results reported at 3 days and at 7 days)
- Mycoplasma testing by ribosomal RNA hybridization assay
Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.
R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.
Product Specific Notices
The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.
TERMS AND CONDITIONS
The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.
R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.
NOTWITHSTANDING ANY OTHER PROVISIONS OF THESE TERMS AND/OR ANY OTHER AGREEMENT BETWEEN R&D SYSTEMS AND PURCHASER FOR THE PURCHASE OF THE PRODUCTS, R&D SYSTEMS' TOTAL LIABILITY TO PURCHASER ARISING FROM OR IN RELATION TO THESE TERMS, AN AGREEMENT BETWEEN THE PARTIES OR THE PRODUCTS, WHETHER ARISING IN CONTRACT, TORT OR OTHERWISE SHALL BE LIMITED TO THE TOTAL AMOUNT PAID BY PURCHASER TO R&D SYSTEMS FOR THE APPLICABLE PRODUCTS. IN NO EVENT WILL R&D SYSTEMS BE LIABLE FOR THE COST OF PROCUREMENT OF SUBSTITUTE GOODS.
Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.
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