The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?

Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.

What is the recommended method for reconstitution of a lyophilized protein or antibody?

Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4oC overnight. Contact Technical Service if product does not go into solution.

Recombinant Human BMP-2 GMP Protein, CF

Name: Recombinant Human BMP-2 GMP Protein, CF
Brand: R&D Systems
SKU: 355-GMP

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 355-GMP

Product Datasheet / COA / SDS

Catalog #

Availability

Size / Price

Qty

Setting the standard in quality research reagents

Key Product Details

Source:	CHO
Accession #:	P12643.1
Structure / Form:	Disulfide-linked homodimer
Applications:	Bioactivity

View Full Technical Details for Recombinant Human BMP-2 GMP Protein, CF

Product Specifications for Recombinant Human BMP-2 GMP Protein, CF

Source	Chinese Hamster Ovary cell line, CHO-derived human BMP-2 protein Gln283-Arg396 Manufactured and tested under current Good Manufacturing Practice (GMP) guidelines.
Purity	>95%, by SDS-PAGE with silver staining, under reducing conditions.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Gln-Ala-Lys-His-Lys-Gln-Arg-Lys-Arg-Leu
Predicted Molecular Mass	13 kDa (monomer)
SDS-PAGE	15-16, kDa, reducing conditions
Activity	Measured by its ability to induce alkaline phosphatase production by ATDC5 mouse chondrogenic cells. Binnerts, M.E. et al. (2004) Biochem. Biophys. Res. Commun. 315(2):272. The ED₅₀ for this effect is 40-200 ng/mL. The specific activity of recombinant human BMP-2 is >5.0 x 10⁵ units/mg, which is calibrated against the human BMP-2 WHO Standard (NIBSC code: 93/574).

Scientific Data Examples for Recombinant Human BMP-2 GMP Protein, CF

	Recombinant Human BMP-2 GMP Protein Bioactivity GMP-grade Recombinant Human BMP-2 (Catalog # 355-GMP) induces alkaline phosphatase production in the ATDC5 mouse chondrogenic cell line. The ED₅₀ for this effect is 40-200 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP BMP-2.
	Recombinant Human BMP-2 GMP Protein SDS-PAGE 1 μg/lane of GMP-grade Recombinant Human BMP-2 (Catalog # 355-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing single bands at 15 kDa and 30 kDa, respectively.

Formulation, Preparation and Storage

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.

355-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in Glycine, Sucrose, Tween® 80 and Glutamic Acid.
Reconstitution	Reconstitute at 100 μg/mL in 4 mM HCl.
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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: BMP-2

Bone morphogenetic protein 2 (BMP-2) is a member of the BMP subgroup of the TGF-beta superfamily. It plays a dominant role in embryonic dorsal-ventral patterning, organogenesis, limb bud formation, and bone formation and regeneration (1, 2). Human BMP‑2 is synthesized as a 396 amino acid (aa) preproprotein that contains a 23 aa signal sequence, a 259 aa prosegment, and a 114 aa mature region (3). Proteolytic removal of the propeptide enables mature BMP-2 to form active disulfide linked homodimers and heterodimers with BMP-7 (2). Mature monomeric BMP-2 is an 18 kDa glycosylated peptide with seven conserved cysteines that form a cystine knot structure (4). Mature human BMP-2 shares 100% aa sequence identity with mouse and rat BMP-2. It shares 85% aa sequence identity with human BMP-4 and less than 51% with other BMPs. BMP-2 signals through heterodimeric complexes composed of a type I receptor (Activin RI, BMPR‑IA, or BMPR‑IB) and a type II receptor (BMP RII or Activin RIIB) (2, 5). BMP-2 induces chondrocyte proliferation, endochondral bone formation, longitudinal bone growth, and bone and cartilage repair (6, 7). It induces ectopic bone formation or calcification by promoting osteogenic and chondrogenic differentiation in mesenchymal cells, stem cells, and vascular smooth muscle cells (2, 8‑10). BMP-2/BMP-7 heterodimers are significantly more potent than BMP-2 homodimers at inducing bone formation in vivo (11). BMP-2 also promotes the maintenance and repair of colonic epithelium, suppresses neuronal dopamine synthesis and release, induces apoptosis in medulloblastoma cells, and is required for cardiac contractility (12‑15).

Item	Value
References	Kishigami, S. and Y. Mishina (2005) Cytokine Growth Factor Rev. 16:265. Chen, D. et al. (2004) Growth Factors 22:233. Wozney, J. et al. (1988) Science 242:1528. Sun, P.D. and D.R. Davies (1995) Annu. Rev. Biophys. Biomol. Struct. 24:269. Sebald, W. et al. (2004) Biol. Chem. 385:697. De Luca, F. et al. (2001) Endocrinology 142:430. Davidson, E.N.B., et al. (2007) Arthritis Res. Ther. 9:R102. Ryoo, H.-M. et al. (2006) Gene 366:51. Kramer, J. et al. (2000) Mech. Dev. 92:193. Li, X. et al. (2008) Atherosclerosis January 5 epub. Zhu, W. et al. (2004) J. Bone Miner. Res. 19:2021. Peiris, D. et al. (2007) Am. J. Physiol. Gastrointest. Liver Physiol. 292:G753. Kano, Y. et al. (2005) Endocrinology 146:5332. Hallahan, A.R. et al. (2003) Nat. Med. 9:1033. Wang, Y.-X. et al. (2007) Cardiovasc. Res. 74:290.
Long Name	Bone Morphogenetic Protein 2
Entrez Gene IDs	650 (Human); 12156 (Mouse); 29373 (Rat)
Alternate Names	BDA2, BMP-2, BMP-2A, BMP2, BMP2ABone morphogenetic protein 2A, SSFSC, bone morphogenetic protein 2

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

Product Specific Notices for Recombinant Human BMP-2 GMP Protein, CF

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.

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.

Citations for Recombinant Human BMP-2 GMP Protein, CF

Please visit our Instrument Citation Database to search our collection of scientific articles that feature our instruments.

FAQs for Recombinant Human BMP-2 GMP Protein, CF

The vial is supposed to contain lyophilized protein but it appears to be empty. Is there anything in it?

Pellets can be dislodged during shipping and become disbursed on the vial wall and in the cap. Centrifuge or tap the vial on the benchtop to return this material to the vial bottom. If this does not reveal a pellet, closely inspect the cone of the vial. Some pellets appear as only a tiny amount of material or as a transparent film due to the original buffer formulation. This is a normal appearance for many proteins. For example, if the product is originally lyophilized from a solvent such as acetonitrile or ethanol, and supplied carrier-free, you may not be able to detect the pellet with the naked eye. This does not mean the vial is empty. Reconstitute the vial as directed. After reconstitution, protein concentration can be tested with a spectrophotometer.
What is the recommended method for reconstitution of a lyophilized protein or antibody?

Unless more specific directions are on the Certificate of Analysis provided with the product, we suggest the following procedure to ensure optimal recovery: 1. Allow the vial and reconstitution buffer to equilibrate to room temperature. 2. Briefly centrifuge the vial to ensure that all lyophiliate is collected at the bottom of the vial. 3. Add the amount of buffer required to achieve the concentration recommended on the product insert. 4. Allow the vial to reconstitute for 15-30 minutes at room temperature with gentle agitation, like on a rocker platform or rotating by hand. Avoid vigorous shaking that can cause foaming and protein denaturation. 5. Aliquot into volumes greater than 20 μL and store as indicated on the product insert. If the vial exhibits flakes or particulates, mix the product for a couple of hours at room temperature and then at 4^oC overnight. Contact Technical Service if product does not go into solution.

Recombinant Human BMP-2 GMP Protein, CF

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

Customers also Viewed

Product Specifications for Recombinant Human BMP-2 GMP Protein, CF

Scientific Data Examples for Recombinant Human BMP-2 GMP Protein, CF

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: BMP-2

Manufacturing Specifications

Product Specific Notices for Recombinant Human BMP-2 GMP Protein, CF

Citations for Recombinant Human BMP-2 GMP Protein, CF

FAQs for Recombinant Human BMP-2 GMP Protein, CF

Product Documents for Recombinant Human BMP-2 GMP Protein, CF

Product Datasheets

COA

Certificate of Analysis

SDS

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