Polycaprolactone (PCL, Mn 70,000), 2-aminoethyl methacrylate hydrochloride, and fluorescamine were purchased from Sigma (Saint Louis, MO, USA). Methanol and tertahydrofuran (THF) were purchased from Duksan Chemical (Seoul, Korea). N,N-dimethyl formamide (DMF) was purchased from Showa Chemical (Japan). E. coli-derived recombinant human bone morphogenetic protein-2 (BMP-2) was donated by Cowellmedi (Busan, Korea). Dulbecco's modified eagle's medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), and penicillin-streptomycin were purchased from Gibco-BRL (Rockville, MD, USA). The cell counting kit-8 (CCK-8) was purchased from Dojindo (Tokyo, Japan). The BMP-2 ELISA kit was purchased from PeproTech Inc. (Rocky Hill, NJ, USA). All chemicals and solvents were used without further purification. MG-63 cells (human osteosarcoma cell line) were purchased from Korea Cell Line Bank (KCLB number 21427, Seoul, Korea).
Bmp 2 elisa kit
Manufactured by Thermo Fisher Scientific
Sourced in United States
The BMP-2 ELISA kit is a laboratory test used for the quantitative measurement of Bone Morphogenetic Protein 2 (BMP-2) levels in various biological samples, such as cell culture supernatants, plasma, and serum. It employs the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify the presence of BMP-2 in the samples.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Mg 63 cells, by Korean Cell Line Bank (1 mentions)
Fluorescamine, by Merck Group (1 mentions)
2 aminoethyl methacrylate hydrochloride, by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Sodium tripolyphosphate tpp, by Merck Group (1 mentions)
Rhbmp2, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 cck 8, by Merck Group (1 mentions)
Vegf elisa kit, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Collagenase p, by Merck Group (1 mentions)
Horse serum, by American Type Culture Collection (1 mentions)
Eagle s minimum essential medium, by American Type Culture Collection (1 mentions)
Dulbecco s modified eagle s medium dmem 30 2002, by American Type Culture Collection (1 mentions)
6 protocols using bmp 2 elisa kit
1
Polycaprolactone-Based Biomaterial for BMP-2 Delivery
2
In vitro BMP-2 Release from CPC and Silk
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To measure the release of BMP-2 from CPC particles and silk sponges in vitro, the BMP-2/CPC particles and BMP-2/silk sponges were respectively put into the vials containing 1 ml PBS solution. The vials were incubated at 37 °C for 7 d. At each time point, the release medium was collected and replaced with an equal amount of fresh PBS. The amount of released BMP-2 was measured using a BMP-2 ELISA kit (BGK8C060, PeproTech, USA). The release profile was defined as the cumulative percentage of released BMP-2 (%, w/w) during the period of incubation. The results were shown in Supplementary Fig. 5 .
3
Chitosan-Poly(dioxanone) Nanoparticles for BMP-2 Delivery
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CH was procured from Aladdin Inc (Shanghai, China). Viscosity average molecular weight and deacetylation degree of CH were determined as 1.19 (±0.15) × 105 and 92.6 (±1.8)% using reported methods [11 (link),12 (link)]. Human recombinant BMP-2 and BMP-2 ELISA Kit were purchased from PeproTech Inc (Rocky Hill, NJ, USA) and Abcan Inc (Shanghai, China), respectively. HA (sodium salt, Mw: 90–110 kDa), GP and sodium tripolyphosphate (TPP) were purchased from Sigma-Aldrich (Shanghai, China). All other reagents and chemicals were of analytical grade and purchased from Sinopharm (Shanghai, China).
CH-PDO copolymers were synthesized by grafting PDO side chains onto the C-6 sites of CH backbone using a group-protection method described elsewhere [13 (link)], and the optimized CH-PDO with a PDO weight percentage of about 30 wt% was used for the preparation of nanoparticles.
CH-PDO copolymers were synthesized by grafting PDO side chains onto the C-6 sites of CH backbone using a group-protection method described elsewhere [13 (link)], and the optimized CH-PDO with a PDO weight percentage of about 30 wt% was used for the preparation of nanoparticles.
4
BMP-2 Encapsulation and Release Kinetics
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To encapsulate BMP-2 into μRB-based scaffold, 5 mg μRBs were rehydrated in 100 μl BMP-2 solution (10 μg/ml). After that, 10 μl μRBs were injected and crosslinked in situ by thrombin, resulting in 10 scaffolds with 100 ng BMP-2 loaded per scaffold.
To assess the release of BMP-2 in vitro, scaffolds containing 100 ng BMP-2 were incubated in 24 well plates with serum containing medium. At each time point, the supernatant was collected, and 1ml fresh medium was added into the well. The amount of released BMP-2 was measured by using BMP-2 ELISA kit (Peprotech, Rocky Hill, NJ).
To assess the release of BMP-2 in vitro, scaffolds containing 100 ng BMP-2 were incubated in 24 well plates with serum containing medium. At each time point, the supernatant was collected, and 1ml fresh medium was added into the well. The amount of released BMP-2 was measured by using BMP-2 ELISA kit (Peprotech, Rocky Hill, NJ).
5
rhBMP-2 Release from Scaffolds
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Sterile PLGA/TCP
and TCP scaffolds were coated with rhBMP-2 (from E. coli; purity >98%; Jiuyuan Genetic Institute of Huadong
Medicine, China) for in vivo and in vitro release study. For in vivo
study, 6 mg of rhBMP-2 was coated on each scaffold (20 mm × 10
mm × 10 mm). Briefly, 5 mL of rhBMP-2 solution (108 mg rhBMP-2
dissolved in 90 mL of 3% w/v gelatin solution prepared in 1% v/v acetic
acid and the final rhBMP-2 concentration was 1.2 mg/mL) was added
to each scaffold, then lyophilized for 24 h, and stored at 2–7
°C for further use. In vitro release kinetics of rhBMP-2 from
the coated PLGA/TCP and TCP scaffolds (200 μg rhBMP-2 per 5
mm × 5 mm × 5 mm scaffold) was analyzed. For rhBMP-2 release
study, the scaffolds (n = 3) were soaked in 1 mL
of 10% phosphate buffer saline (PBS; pH 7.4, at 37 °C) in a closed
centrifuge tube and agitated at 60 rpm. The total volume of the medium
was collected after 3, 6, 9, 24, 48, 72, 120, 168, 336, and 504 h
and thereafter analyzed by enzyme linked immunosorbent assay (ELISA)
using a BMP-2 Elisa kit (Peprotech, U.S.A.) to calculate the percentage
of rhBMP-2 release by dividing the measured contents (from a standard
curve) with loaded amount of rhBMP-2. An equal volume of fresh medium
was replaced every time after withdrawal of medium.
and TCP scaffolds were coated with rhBMP-2 (from E. coli; purity >98%; Jiuyuan Genetic Institute of Huadong
Medicine, China) for in vivo and in vitro release study. For in vivo
study, 6 mg of rhBMP-2 was coated on each scaffold (20 mm × 10
mm × 10 mm). Briefly, 5 mL of rhBMP-2 solution (108 mg rhBMP-2
dissolved in 90 mL of 3% w/v gelatin solution prepared in 1% v/v acetic
acid and the final rhBMP-2 concentration was 1.2 mg/mL) was added
to each scaffold, then lyophilized for 24 h, and stored at 2–7
°C for further use. In vitro release kinetics of rhBMP-2 from
the coated PLGA/TCP and TCP scaffolds (200 μg rhBMP-2 per 5
mm × 5 mm × 5 mm scaffold) was analyzed. For rhBMP-2 release
study, the scaffolds (n = 3) were soaked in 1 mL
of 10% phosphate buffer saline (PBS; pH 7.4, at 37 °C) in a closed
centrifuge tube and agitated at 60 rpm. The total volume of the medium
was collected after 3, 6, 9, 24, 48, 72, 120, 168, 336, and 504 h
and thereafter analyzed by enzyme linked immunosorbent assay (ELISA)
using a BMP-2 Elisa kit (Peprotech, U.S.A.) to calculate the percentage
of rhBMP-2 release by dividing the measured contents (from a standard
curve) with loaded amount of rhBMP-2. An equal volume of fresh medium
was replaced every time after withdrawal of medium.
6
Engineered Bone Regeneration Scaffold
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Gelatin from bovine skin Type B with ≈ 225 bloom strength, Cell Counting Kit-8 (CCK-8) and Collagenase P were purchased from Sigma-Aldrich (Spain). mTG derived from Streptoverticillium mobaraense with a specific activity of 100 U/g was kindly supplied by Ajinomoto Foods Europe (France). Mouse L-929 fibroblasts and mouse D1 ORL UVA Mesenchymal Stem Cells (MSCs) cell lines, Dulbecco's Modified Eagle's Medium (DMEM 30-2002) , horse serum and Eagle's Minimum Essential Medium (EMEM 30-2003) culture mediums were obtained from ATCC (Spain). MG63 cells were immortalized osteoblasts isolated from a 14 years-old caucasian male osteosarcoma. Trypsin, Hoechst 33342, fetal bovine serum (FBS), Phosphate Buffered Saline (PBS) pH=7.4 (1X) and Penicillin-Streptomycin were purchased from Fisher Scientific (Spain). rhVEGF was kindly supplied by Agrenvec (Spain). Finally, VEGF ELISA kit, BMP-2 ELISA kit and rhBMP-2 were obtained from Peprotech (UK).
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