Fetal bovine serum (FBS), normal goat serum (NGS), Dulbecco’s modified Eagle’s medium (DMEM; #1200-046), and 10X MEM (11430-030) were purchased from Life TechnologiesTM. Amphotericin B (BP2645) and GlutaMaxTM Supplement (35050-061) were obtained from Gibco®, and Pen Strep (15140-122) was obtained from Fisher Scientific. PureCol® type I bovine collagen (3 mg/mL) was obtained from Advanced Biomatrix. Antibodies against p53 (SC6243), PUMA (SC374223) and β-actin (SC8432) were obtained from Santa Cruz Biotechnology®, Inc. The antibody against α-SMA (SAB5500002) was purchased from Sigma, Inc.
Purecol bovine type 1 collagen
Manufactured by Advanced BioMatrix
Sourced in United States
PureCol is a high-purity, bovine-derived type I collagen product. It is a natural, fibrillar collagen that maintains its native structure. PureCol can be used in a variety of research and cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fibronectin, by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Propidium iodide, by Thermo Fisher Scientific (2 mentions)
Recombinant human hgf, by R&D Systems (2 mentions)
Collagenase type 1, by Merck Group (2 mentions)
Crizotinib, by Merck Group (2 mentions)
E4500 digital camera, by Nikon (2 mentions)
Mz6 modular stereomicroscope, by Leica camera (2 mentions)
Sylgard 184 silicone elastomer, by Dow (2 mentions)
D21490, by Thermo Fisher Scientific (2 mentions)
Ec plan neofluar 10 0.3 dici objective lens, by Zeiss (2 mentions)
Antibody against α sma sab5500002, by Merck Group (1 mentions)
Amphotericin b bp2645, by Thermo Fisher Scientific (1 mentions)
Glutamaxtm supplement, by Thermo Fisher Scientific (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Nsc23766, by Santa Cruz Biotechnology (1 mentions)
Recombinant human tgf β1, by Cell Signaling Technology (1 mentions)
Complete dmem f12, by Thermo Fisher Scientific (1 mentions)
Cabozantinib, by Selleck Chemicals (1 mentions)
R428 bgb324, by Selleck Chemicals (1 mentions)
17 protocols using purecol bovine type 1 collagen
1
Fetal Bovine Serum Collagen Tissue Culture
2
Cell Adhesion Assay with EV1 and Collagen
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
2.5 μg/cm2 EV1, 5 μg/cm2 Pure Col type I bovine collagen (Advanced BioMatrix), or 0.1 mg/ml poly-L-lysine in PBS was incubated on cell culture plate overnight at +4 °C, and plates were blocked with 0.1% BSA in PBS for 1 hour at +37 °C. Cells were grown in low serum (0.5%) overnight, and where indicated, treated with inhibitors, shRNA, or EDTA. Inhibitors for PKC (safingol, 10 µM, 30 min, Calbiochem) and Rac1 (NSC23766, 100 µM, 1 h, Santa Cruz) were added prior cell plating and kept during cell adhesion. 5 mM EDTA was added 10 minutes before plating the cells and kept during cell adhesion. When detaching the cells for the experiment, trypsin-EDTA was allowed to completely detach the cells. Trypsin was inhibited with equal volume of 1 mg/ml trypsin inhibitor, and cells were pelleted and washed, and then re-suspended in serum free DMEM. Cells were allowed to attach the virus coated surface for indicated time (15–60 min) and lysed for Western blot analysis. Control samples for Western blot (0 min) were pelleted and lysed without cell plating.
3
3D Collagen Gel Fibroblast Contraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three-dimensional collagen gels were prepared by mixing 16 mL of chilled collagen solution (PureCol® Type I bovine collagen; Advanced BioMatrix, Sand Diego, CA), with 2 mL of sterile 10X PBS. While keeping the solution on ice, the pH was adjusted to 7.4 using sterile 0.1 M NaOH, and the volume was brought up to 20 mL with sterile water. Gels were cast in a 24-well plate by adding 600 µL solution to each well and allowing them to solidify at 37 °C, 5% CO2 overnight. The following day, P0 fibroblasts cultured on 5 kPa elastic surfaces were passaged and seeded on the collagen matrices at a density of 1.5 × 104 cells per well and allowed to adhere for 24 h. The gels were then released from the wells using a circular cutting tool and immediately infected with their corresponding adenoviral vector, or treated with 4 ng/mL recombinant human TGF-β1 (Cell Signaling, #8915) as a positive control. Images were taken immediately following treatment, and subsequently every 24 h for a total of 72 h post-treatment. Gel contraction was estimated by measuring the surface area of the top of the gel using ImageJ image processing software [44 (link)].
4
Stacks Microfluidic Platform for 3D Cell Culture
5
Stacks Microfluidic Platform for 3D Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Description of the Stacks microfluidic platform is previously described in detail 27 (link) . 24-well polystyrene microdevices (dimensions: 75 mm x 50 mm) were fabricated by injection molding and sterilized via sonication in isopropanol. Collagen hydrogels (2.0 mg/ml) were prepared and a 4.5 µL volume was suspended within each microwell of the device (thickness ~1.2 mm), creating an open-air culture system compatible for two-photon imaging. The hydrogel was generated from a mixture of 6 µL 10X PBS, 14 µL sterile water, 6 µL NaOH (Sigma), 160 µL Bovine collagen type I (PureCol, Advanced BioMatrix), 3 µL fibronectin (Sigma) and 50 µL cell type-specific medium (RPMI1640 -mouse; complete DMEM/F12 (Gibco) -human). Collagen hydrogels were polymerized by incubation at 37⁰C for at least 6 hours, prior to sequential seeding of breast carcinoma cells and macrophages (~1000 cells/μL each) on the opposing ends of the collagen layer.
6
Immune Cell Interactions in Collagen Matrices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen matrices were prepared as previously described [24 (link)]. Briefly, CAFs (1 × 105), 20 PANC-1 spheroids and PBMCs (1 × 106) were embedded in PureCol bovine type I collagen (Advanced Biomatrix, San Diego, CA, USA, cat. no. 5005) mixed with 5 × DMEM, NaHCO3, FBS and PEST to get a final concentration of 2 mg/mL of collagen I.
On day 5, PBMCs were released from the matrices by mechanical disaggregation using a GentleMACSTM Dissociator and analyzed by flow cytometry.
On day 5, PBMCs were released from the matrices by mechanical disaggregation using a GentleMACSTM Dissociator and analyzed by flow cytometry.
7
Evaluation of Anti-Tumor Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PureCol bovine type I collagen was purchased from Advanced Biomatrix, Inc. (San Diego, CA, USA, #5005-100 ml). Unless specified otherwise, all cell culture components were purchased from Hyclone Laboratories, Inc. (Omaha, NE, USA). DMEM was purchased from Corning Mediatech (Manassas, VA, USA, #10-017-CV). Crizotinib was purchased from MilliporeSigma (Temecula, CA, USA, #PZ0191). Cabozantinib (#S1119), BMS-777607 (#S1561), and R428 (BGB324, #S2841) were purchased from Selleck Chemicals, Houston, TX, USA. Collagenase, type I was purchased from MilliporeSigma (Temecula, CA, USA, #234153). Recombinant human HGF was purchased from R&D Systems (Minneapolis, MN, USA, #294-HG/CF). Propidium iodide was purchased from Invitrogen (Carlsbad, CA, USA, #P3566).
8
Collagen Gel Contraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen gels were prepared by mixing fibroblasts with PureCol bovine type I collagen (Advanced Biomatrix), 5x DMEM, 0.1 M NaOH, and distilled water (final concentrations were 1.7 mg/ml PureCol, 1x DMEM, and 3 mM NaOH) in the presence of 1,25(OH)2D3, Wnt3A, both, or vehicle. The mixture was seeded in 24-well cell culture plates and allowed to polymerize for 1 h at 37 °C. Then, culture medium with the corresponding treatments (1,25(OH)2D3, Wnt3A, both, or vehicle) was added. After 24 h and to initiate gel contraction (time 0), gels were gently released from the 24-well plates and transferred into 6-well plates containing culture medium with 1,25(OH)2D3, Wnt3A, both, or vehicle. Gels were photographed at 0 and 96 h with a Leica DFC550 digital camera mounted in a Leica S6D stereomicroscope and the gel areas were measured with ImageJ software. Images were processed using Adobe Photoshop CC software. All experiments were performed using triplicates.
9
Collagen Gel Contraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen gels were prepared by mixing fibroblasts with PureCol bovine type I collagen (Advanced Biomatrix, San Diego, California, USA), 5x Dulbecco's modified Eagle's medium (DMEM), 0.1 M NaOH and distilled water (final concentrations: 1.7 mg/mL PureCol, 1x DMEM and 3 mM NaOH) in the presence of 100 nM 1,25(OH)2D3 or vehicle. The mixture was seeded in 24-well cell culture plates and allowed to polymerise. Then, culture medium with 100 nM 1,25(OH)2D3 or vehicle was added. After 24 h and to initiate gel contraction (time 0), gels were released from the 24-well plates and transferred into 6-well plates containing culture medium with 100 nM 1,25(OH)2D3 or vehicle. At the indicated times, images were taken with an E4500 digital camera (Nikon, Tokyo, Japan) mounted in a MZ6 modular stereomicroscope (Leica, Wetzlar, Germany), and gel area was measured with ImageJ (National Institutes of Health, Bethesda, Maryland, USA). Images were processed using Adobe Photoshop CS6 (San Jose, California, USA). Experiments were performed using triplicates.
10
Collagen-based Cell Culture System
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PureCol bovine type I collagen was purchased from Advanced Biomatrix, Inc. (San Diego, CA, USA, #5005–100 ml). Unless specified otherwise, all cell culture components were purchased from Hyclone Laboratories, Inc. (Omaha, NE, USA). DMEM and 10X DMEM were purchased from Gibco (#11,430–030). Collagenase, type I was purchased from MilliporeSigma (Temecula, CA, USA, #234,153). Recombinant human HGF was purchased from R&D Systems (Minneapolis, MN, USA, #294-HG/CF). Recombinant human HAI-1 was purchased from SinoBiological (Houston, TX, USA, #11,742-H08H). Propidium iodide was purchased from Invitrogen (Carlsbad, CA, USA, #P3566). Sodium Hydroxide 10 Normal was purchased from VWR Chemicals BDH (Radnor, PA, USA, #BDH3247-1). Crizotinib was purchased from MilliporeSigma (Temecula, CA, USA, #PZ0191). Cetuximab was purchased from Eli Lilly and Company. ZFH7116, VD2173, MM3122, VD4162, and VD5064 were generated in the lab of James Janetka at Washington University in St. Louis, MO.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!