Recombinant Human LR3 IGF-I/IGF-1 GMP Protein, CF

Catalog #: 8335D-GMP Datasheet / COA / SDS

Animal-Free

Catalog #	Availability	Size / Price	Qty
8335D-GMP-200
8335D-GMP-01M

Recombinant Human LR3 IGF-I/IGF-1 GMP Protein Bioactivity

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Product Details

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human LR3 IGF-I/IGF-1 GMP Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. The result of the major peak from mass spectrometry analysis is 9111 Da, which corresponds to the calculated molecular mass of 9118 Da.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

Activity

Measured in a serum-free cell proliferation assay using MCF‑7 human breast cancer cells. Karey, K.P. et al. (1988) Cancer Research 48:4083. The ED₅₀ for this effect is 0.3-1.5 ng/mL. IGFBP-3 does not inhibit its activity.

Source

E. coli-derived human IGF-I/IGF-1 protein

MFPAMPLSSLFVN	Human LR3 IGF-I/IGF-1 (Gly49-Ala118 (Glu51Arg)) Accession # P05019
N-terminus	C-terminus

Produced using non-animal reagents in an animal-free laboratory. Manufactured and tested under cGMP guidelines.

Accession #

P05019.1

N-terminal Sequence
Analysis

Met-Phe-Pro-Ala-Met-Pro-Leu-Ser-Ser-Leu

Predicted Molecular Mass

9 kDa

SDS-PAGE

8 kDa, reducing conditions

Product Datasheets

8335D-GMP

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

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.

What formulation is right for me?

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.

8335D-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Acetic Acid.
Reconstitution	Reconstitute at 100-200 μg/mL in sterile 100 mM Acetic Acid
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Scientific Data

Bioactivity

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GMP-grade Recombinant Human LR3 IGF-I/IGF-1 (Catalog # 8335D‑GMP) stimulates cell proliferation in a serum-free assay using the MCF‑7 human breast cancer cell line. The ED₅₀ for this effect is 0.3‑1.5 ng/mL.

SDS-PAGE

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2 μg/lane of GMP-grade Recombinant Human LR3 IGF-I/IGF-1 (Catalog # 8335D-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing R and NR bands at 8 kDa.

Reconstitution Calculator

Background: IGF-I/IGF-1

Insulin-like Growth Factor 1 (IGF-1 or IGF-I), also known as Somatomedin C, is the dominant effector of Growth Hormone (GH) and is structurally homologous to Proinsulin. Human IGF-1 is synthesized as two precursor isoforms with N- and alternative C‑terminal propeptides (1). These isoforms are differentially expressed by various tissues (1). The 7.6 kDa mature IGF‑1 is identical between isoforms and is generated by proteolytic removal of the N- and C-terminal regions. Mature human IGF-1 shares 94% and 96% amino acid (aa) sequence identity with the mouse and rat orthologs, respectively (2). GH stimulates the production of IGF-1 in most tissues (3). Hepatocytes produce circulating IGF-1, while local IGF-1 is produced by many other tissues in which it has paracrine effects (1). IGF-1 induces the proliferation, migration, and differentiation of a wide variety of cell types during development and postnatally (4, 5). IGF-1 regulates glucose, fatty acid, and protein metabolism, steroid hormone activity, and cartilage and bone metabolism (6-11). It plays an important role in muscle regeneration and tumor progression (1, 12, 13). IGF-1 binds IGF1R, IGF2R, and the Insulin Receptor, although its effects are mediated primarily by IGF1R (14). IGF-1 also binds with strong affinity to IGF binding proteins (IGFBPs), which regulate the availability and biological activities of IGF-1 (15, 16).

Long R3 IGF-1 (LR3 IGF-1) is a 9.2 kDa synthetic analog of IGF-1 that is generated by modifying the aa sequence for mature human IGF-1. These modifications include the substitution of an Arg for Glu at position 3 of the mature IGF-1 sequence and the addition of a thirteen aa N-terminal extension, which is derived from methionyl porcine Growth Hormone (17). These aa changes generate a protein that is still capable of binding to IGF-1 and Insulin receptors, but shows considerably lower affinity binding to IGFBPs compared to wild-type IGF-1 (17, 18). As a result, LR3 IGF-1 has an increased half-life and displays increased biological potency compared to IGF-1 (17-22).

References

Philippou, A. et al. (2007) In Vivo 21:45.
Sandberg-Nordqvist, A.C. et al. (1992) Brain Res. Mol. Brain Res. 12:275.
Berryman, D.E. et al. (2013) Nat. Rev. Endocrinol. 9:346.
Guvakova, M.A. (2007) Int. J. Biochem. Cell Biol. 39:890.
Sadagurski, M. and M.F. White (2013) Endocrinol. Metab. Clin. North Am. 42:127.
Clemmons, D.R. (2006) Curr. Opin. Pharmacol. 6:620.
Bluher, S. et al. (2005) Best Pract. Res. Clin. Endocrinol. Metab. 19:577.
Garcia-Segura, L.M. et al. (2006) Neuroendocrinology 84:275.
Malemud, C.J. (2007) Clin. Chim. Acta 375:10.
Ling, P.R. et al. (1995) Am. J. Clin. Nutr. 61:116.
Sheng, M.H. et al. (2014) J. Bone Metab. 21:41.
Samani, A.A. et al. (2007) Endocrine Rev. 28:20.
Gallagher, E.J. et al. (2010) Endocr. Pract. 16:864.
LeRoith, D. and S. Yakar (2007) Nat. Clin. Pract. Endocrinol. Metab. 3:302.
Denley, A. et al. (2005) Cytokine Growth Factor Rev. 16:421.
Duan, C. and Q. Xu (2005) Gen. Comp. Endocrinol. 142:44.
Francis, G.L. et al. (1992) J. Mol. Endocrinol. 8:213.
Voorhamme, D. and C.A. Yandell (2006) Mol. Biotechnol. 34:201.
Tomas, F.M. et al. (1993) Biochem. J. 291:781.
Tomas, F.M. et al. (1996) J. Endocrinol. 150:77.
Tomas, F.M. et al. (1997) J. Endocrinol. 155:377.
von der Thüsen, J.H. et al. (2011) Am. J. Pathol. 178:924.

Long Name

Insulin-like Growth Factor I/Insulin-like Growth Factor 1

Entrez Gene IDs

3479 (Human); 16000 (Mouse); 24482 (Rat)

Alternate Names

IBP1; IGF1; IGF-1; IGF1A; IGFI; IGF-I; IGF-IA; IGF-IB; insulin-like growth factor 1 (somatomedin C); insulin-like growth factor 1; insulin-like growth factor I; insulin-like growth factor IA; insulin-like growth factor IB; Mechano growth factor; MGF; Somatomedin A; Somatomedin C; somatomedin-C

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

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

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

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 testing according to USP
Host Cell Protein testing performed by ELISA
Mycoplasma testing by ribosomal RNA hybridization assay

Additional testing and documentation requested by the customer can be arranged at an additional cost.

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 cell therapy applications. They are not for in vivo use. Please read the following End User Terms prior to using this product.

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.

Production

All molecular biology procedures use animal-free media and dedicated labware.
Dedicated fermentors are utilized in committed animal-free areas.

Purification

Protein purification columns are animal-free.
Bulk proteins are filtered using animal-free filters.
Purified proteins are stored in animal-free containers in a dedicated cold storage room.

Quality Assurance

Low Endotoxin Level.
No impairment of biological activity.
High quality product obtained under stringent conditions.

Please read our complete Animal-Free Statement.

Product Specific Notices

END USER TERMS OF USE OF PRODUCT

The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

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.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal.

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems' GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

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.

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Full details of R&D Systems' Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal.