Human gingival fibroblasts were treated with IL + 0.1N + 50HGF and anti-human HGF antibody (anti-HGF, R&D system) for 48 h, after which the culture supernatants were collected and used to measure the level of type I collagen.
Anti hgf
Manufactured by R&D Systems
Sourced in United States
Anti-HGF is a recombinant protein that acts as an antagonist for Human Hepatocyte Growth Factor (HGF). It is used for research purposes to study the biological functions and signaling pathways of HGF.
Automatically generated - may contain errors
Lab products found in correlation
Amd3100, by Merck Group (1 mentions)
Pf184, by Bio-Techne (1 mentions)
Amd3100, by Sanofi (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Anti e cadherin, by BD (1 mentions)
Anti p met, by Cell Signaling Technology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Enhanced chemi luminescence (ecl), by Cytiva (1 mentions)
Anti met, by R&D Systems (1 mentions)
Anti p gab1, by Cell Signaling Technology (1 mentions)
Anti vimentin, by Cell Signaling Technology (1 mentions)
Anti p erk1 2, by Cell Signaling Technology (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Transwell system, by Corning (1 mentions)
Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Gm csf, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Hepatocyte growth factor (hgf), by R&D Systems (1 mentions)
Roswell park memorial institute (rpmi), by Corning (1 mentions)
5 protocols using anti hgf
1
Gingival Fibroblast Collagen Regulation
2
Inhibiting Malaria Sporozoite Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For in vitro experiments, AMD3100 (1 µM; Sigma-Aldrich), anti-HGF (20 µg/ml; R&D Systems), PF184 (500 nM; Tocris), or FR182024 (10 µM; Merck Millipore) was added to cell culture media 4 h before infection. Each inhibitor and medium were changed at 0, 4, and 24 h after infection.
For in vivo experiments, AMD3100 (10 mg/kg) was intraperitoneally injected into mice at 24 and 12 h before infection. After intravenous infection of sporozoites, mice were injected with AMD3100 at 0, 12, and 24 h after infection. According to the manufacturer’s protocol for AMD3100 (also known as Plerixafor; Genzyme Plerixafor Injection Investigator's Brochure, 2009, version 13, May 27, 2009), a half-life of AMD3100 was previously shown to be ∼5–6 h. Also, the manufacturer reported that the effect of AMD3100 in vivo continues four half-life times (∼20–24 h). In addition, our preliminary experiments to test how often normal mice can be treated with AMD3100 suggested that mice treated with AMD3100 (10 mg/kg) every 6 h become unhealthy. Therefore, we decided to administer AMD3100 treatment in mice every 12 h; treatments were performed at 24 h and 12 h before infection and at 0 h, 12 h, and 24 h after infection.
For in vivo experiments, AMD3100 (10 mg/kg) was intraperitoneally injected into mice at 24 and 12 h before infection. After intravenous infection of sporozoites, mice were injected with AMD3100 at 0, 12, and 24 h after infection. According to the manufacturer’s protocol for AMD3100 (also known as Plerixafor; Genzyme Plerixafor Injection Investigator's Brochure, 2009, version 13, May 27, 2009), a half-life of AMD3100 was previously shown to be ∼5–6 h. Also, the manufacturer reported that the effect of AMD3100 in vivo continues four half-life times (∼20–24 h). In addition, our preliminary experiments to test how often normal mice can be treated with AMD3100 suggested that mice treated with AMD3100 (10 mg/kg) every 6 h become unhealthy. Therefore, we decided to administer AMD3100 treatment in mice every 12 h; treatments were performed at 24 h and 12 h before infection and at 0 h, 12 h, and 24 h after infection.
3
Paracrine Effects of hASCs on Cardiac Myofibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Conditioned medium of hASCs was given to cardiac fibroblasts following 5-day culture on PA hydrogels, to determine the paracrine effects of hASCs on cardiac myofibroblast differentiation. The treatment lasted for 3 days. To explore the potential role of HGF on anti-fibrotic activity of conditioned medium of hASCs, the conditioned medium was incubated with 3 ng/mL neutralizing antibody for HGF (anti-HGF) (R & D System, Minneapolis, MN, USA) at 37 °C for 1 hour prior to being given to cardiac fibroblasts. After 3 day-treatment, myofibroblast differentiation evaluation of these cells was performed. The concentration of HGF in conditioned medium of hASCs was determined by ELISA (described later).
4
Immunoblotting Analysis of Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoblotting was performed using standard procedures. Membranes were blocked with 5% non-fat milk for 1 hour at RT, and incubated with primary antibodies overnight at 4°C. Primary antibodies used were anti-HGF (R&D Systems), anti-MET, anti-p-MET, anti-p-GAB1, anti-p-AKT, anti-p-ERK1/2, anti-vimentin (Cell Signaling Technology), anti-p-STAT3 (Upstate Cell Signaling Solutions), anti-E-cadherin (BD Transduction Laboratories), anti-β-actin (Sigma). Following washing with TBS-T buffer, membranes were incubated with horseradish peroxidase-conjugated secondary antibodies for 1 hour and then washed with TBS-T. Immunoblots were developed using the chemiluminescent detection system with ECL (Amersham). Protein loading was normalized by probing blots for the expression of β-actin.
5
Modulation of Monocyte Differentiation by UC-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly isolated monocytes (CD14+) were cultured at a concentration of 4.0 × 105 cells/well in 12-well plates containing 1 ml RPMI (Invitrogen Corp., Paisley, UK) with 100 U/ml penicillin and streptomycin (P/S; Invitrogen Corp), L-glutamine (Invitrogen Corp., Paisley, UK), 10% FBS and the growth factors IL4 (50 ng/ml, PeproTech, USA) and GM-CSF (50 ng/ml, PeproTech, USA) for 5 days, resulting in the generation of immature DCs (iDCs; CD14-/CD1a+). After the 5-day incubation, to gain mature DCs (mDCs), LPS (100 ng/ml, Sigma-Aldrich, USA) was added to stimulate the cells for 48 h.
To examine the effect of UC-MSCs on monocyte differentiation into mDCs, UC-MSCs treated with mitomycin were cultured at a concentration of 2.0 × 105 cells/well in 12-well plates containing 1 ml RPMI for 24 h, and then monocytes, IL4 and GM-CSF were added with direct cell-cell contact or in a transwell system (pore size 0.4 μM; Corning Inc., Lowell, MA, USA) at a MSC-monocyte ratio of 1:10 for 5 days and then LPS was added to stimulate the cells for 48 h. After 7-day incubation, monocytes were the separated from the UC-MSCs by spinning the cells in suspension and then washing them.
To examine the effect of some soluble factors, IL10 (50 ng/ml), IL6 (50 ng/ml), HGF (50 ng/ml), anti-IL10 (5 μg/ml), anti-HGF or anti-IL6 (5 μg/ml) (R&D Systems Europe Ltd., Abingdon, UK) was added every 3 days.
To examine the effect of UC-MSCs on monocyte differentiation into mDCs, UC-MSCs treated with mitomycin were cultured at a concentration of 2.0 × 105 cells/well in 12-well plates containing 1 ml RPMI for 24 h, and then monocytes, IL4 and GM-CSF were added with direct cell-cell contact or in a transwell system (pore size 0.4 μM; Corning Inc., Lowell, MA, USA) at a MSC-monocyte ratio of 1:10 for 5 days and then LPS was added to stimulate the cells for 48 h. After 7-day incubation, monocytes were the separated from the UC-MSCs by spinning the cells in suspension and then washing them.
To examine the effect of some soluble factors, IL10 (50 ng/ml), IL6 (50 ng/ml), HGF (50 ng/ml), anti-IL10 (5 μg/ml), anti-HGF or anti-IL6 (5 μg/ml) (R&D Systems Europe Ltd., Abingdon, UK) was added every 3 days.
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!