Nasal tissues from 21 participants with adequate specimens, including seven smokers with asthma and CRS, nine non-smokers with asthma and CRS, and five asthmatic non-smokers without CRS were processed for western blot analyses of MMPs. Total protein was prepared using a 1% IGEPAL lysis buffer (Sigma-Aldrich. St. Louis, MO, USA). Cellular proteins (10 μg) were fractionated by SDS-PAGE and electroblotted onto polyvinylidene difluoride membranes. Primary antibodies including MMP-1 (1:200; Millipore, Billerica, MA, USA), MMP-9 (1:1000; CST, MA, USA), MMP-12 (1:100; Santa Cruz Biotechnology, Dallas, TX, USA), and β-actin (1:20,000; Millipore, Billerica, MA, USA) were used to assess differences in protein levels by enhanced chemiluminescence. Protein bands were visualized using a gel documentation system (Alpha Innotech, San Leandro, CA. USA). Relevant band intensities were quantified via densitometric analyses and normalized to β-actin.
Mmp 1
Manufactured by Merck Group
Sourced in United States, Italy
MMP-1 is a laboratory instrument designed for the detection and quantification of Matrix Metalloproteinase-1 (MMP-1), an enzyme involved in the breakdown of the extracellular matrix. The core function of this product is to provide accurate and reliable measurements of MMP-1 levels in various samples, such as cell culture media or biological fluids. The instrument utilizes advanced detection technologies to enable researchers to study the role of MMP-1 in biological processes and disease states.
Automatically generated - may contain errors
Lab products found in correlation
Uvband, by Eppendorf (3 mentions)
Amplified opti 4cn, by Bio-Rad (3 mentions)
Mmp 9, by Merck Group (3 mentions)
β actin, by Merck Group (2 mentions)
Amplified opti 4cn substrate, by Bio-Rad (2 mentions)
Col 3, by Merck Group (2 mentions)
Mmp 2, by Merck Group (2 mentions)
Mmp 13, by Merck Group (2 mentions)
Mmp 8, by Merck Group (2 mentions)
Gel documentation system, by Bio-Techne (1 mentions)
Igepal lysis buffer, by Merck Group (1 mentions)
Mmp 12, by Santa Cruz Biotechnology (1 mentions)
Elastin, by Merck Group (1 mentions)
Elastin, by Abcam (1 mentions)
Chemiimager analyzer system, by Bio-Techne (1 mentions)
Mmp 1, by Abcam (1 mentions)
Supersignal west dura extended duration substrate, by Thermo Fisher Scientific (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Bio-Rad (1 mentions)
Mmp 3, by Merck Group (1 mentions)
11 protocols using mmp 1
1
Airway MMP Regulation in Asthma and CRS
2
Quantitative Analysis of Collagen I and MMP-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen type I (COL-I) and matrix metalloproteinase (MMP)-1 protein levels secreted by tenocytes in serum-free cell supernatants were analyzed by slot blot analysis, as previously detailed [18 (link)]. Membranes were incubated for 1 h at room temperature with primary monoclonal antibodies to COL-I (1:1000 in TBST) (Sigma-Aldrich, Milan, Italy) or MMP-1 (1 µg/mL in TBST) (Millipore, Milan, Italy). Immunoreactive bands were revealed by the Amplified Opti-4CN substrate (Amplified Opti-4CN, Bio Rad, Segrate, Milan, Italy) and quantification was obtained after densitometric scanning of immunoreactive bands (UVBand, Eppendorf, Italy).
3
Slot Blot Analysis of Tenocyte Secretions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen type I and III (COL-I, COL-III), matrix metalloproteinase (MMP)-1 protein levels secreted by tenocytes were assessed in duplicate samples by Slot blot in serum free cell culture medium, as previously detailed [21 ]. Membranes were incubated for 1 h at room temperature in monoclonal antibody to COL-I (1:1000 in TBST) (Sigma-Aldrich, Milan, Italy), COL-III (1:2000 in TBST) (Sigma-Aldrich, Milan, Italy), MMP-1 (1 µg/mL in TBST) (Millipore, Milan, Italy). Immunoreactive bands, revealed by the Amplified Opti-4CN substrate (Amplified Opti-4CN, Bio Rad, Italy), were scanned densitometrically (UVBand, Eppendorf, Italy).
4
Quantifying Collagen and MMP-1 in Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Collagen type I and III (COL-I and COL-III) and matrix metalloproteinase (MMP)-1 protein levels secreted by muscle fibroblasts were analysed by Slot Blot in serum-free cell supernatants as previously described [67 (link)]. After blocking, membranes were incubated for 1 h at room temperature with primary polyclonal antibodies to COL-I (1:1000 in 1X Tris-Buffered Saline plus 0.1% Tween 20, TBST) (Sigma-Aldrich, Milan, Italy), COL-III (1:1000 in TBST) (Sigma-Aldrich), or MMP-1 (1 μg/mL in TBST) (Millipore, Milan, Italy). After 1 h incubation with HRP-conjugated antibody (1:20,000 in TBST), immunoreactive bands were revealed by the Amplified Opti-4CN substrate (Amplified Opti-4CN, Bio Rad, Segrate, Milan, Italy) and quantified by densitometric scanning (UVBand, Eppendorf, Milan, Italy).
5
Protein Expression Analysis in HaCaT Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the cells were treated in the same manner as the method for measuring cell viability, total protein from HaCaT cell lysates were subjected to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis using 12% gels. Protein bands were then transferred to PVDF membranes (BioRad, Hercules, CA, USA), which were blocked with 5% skim milk in PBS then incubated with a 1:1000 v/v dilution of primary antibodies against COX-2, PGE2, PCOL1 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), β-actin (Cell Signaling, Danvers, MA, USA), MMP-1 and Elastin (Abcam, Cambridge, MA, USA) in PBS with 1% skim milk overnight at 4°C. The blots were then incubated with peroxidase-conjugated goat anti-rabbit IgG (PGE2, β-actin, MMP-1, Elastin) and were then incubated with peroxidase-conjugated goat anti-mouseIgG (COX-2, PCOL1) (1:10,000 v/v, Millipore, CA, USA) for 1 hr. The immunoreactions were visualized with SuperSignal West Dura Extended Duration Substrate (Thermo Scientific, CA, USA) on a ChemiImager analyzer system (Alpha Innotech, San Leandro, CA, USA).
6
Inhibition of MMP and ADAM Proteases
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
KP‐457 and GM‐6001 were tested for their ability to inhibit MMP‐ and ADAM‐catalyzed cleavage of substrates in a fluorescence‐based assay. Human ADAM17, ADAM10, and MMP17 were from R&D Systems (Minneapolis, MN, http://www.rndsystems.com ), and human MMP1, MMP2, MMP3, MMP8, MMP9, MMP13, and MMP14 were from EMD Millipore. MMP1, MMP2, MMP8, MMP9, MMP13, and MMP17 were activated using p‐aminophenylmercuric acetate (Sigma‐Aldrich) before testing. Fluorogenic substrates for measuring activity of ADAM10 and ADAM17 [Nma‐LAQAVRSSK(Dnp)r‐NH2, based on the cleavage site of TNF‐α]; MMP1, MMP9, MMP13, and MMP14 [Dnp‐P‐Cha‐GC(Me)HAK(N‐Me‐Abz)‐NH2]; MMP3 [MOCAc‐RPKPVE‐Nva‐WRK(Dnp)‐NH2]; and MMP2, MMP8, and MMP17 [MOCAc‐PLGL‐A2pr(Dnp)‐AR‐NH2] were all provided by Peptide Institute (Osaka, Japan, https://www.peptide.co.jp/en ) and used as substrates. In addition, inhibitory activities were measured using LC/MS/MS with a GPIbα‐based substrate peptide (KKTIPELDQPPKLRGVLQGHLESSRNDPFLHPDF), a C terminal‐based standard peptide (VLQGHLESSRNDPFLHPDF), and an internal standard peptide (VTTGKGQDHSPFWGF). These peptides were synthetized by Scrum (Tokyo, Japan, http://www.scrum‐net.co.jp/english/top_en.htm ).
7
Placental Amniotic Membrane Tissue Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This work followed the tenets of the Declaration of Helsinki involving human subjects and received approval from our institutional review board. Human AM tissue (ie, the innermost layer of the placental membrane) was obtained with informed consent at the time of elective cesarean section from mothers when human immunodeficiency virus, syphilis, and hepatitis B and C had been excluded by serological tests. Separation of the AM was performed by cutting the tissue samples approximately 2 cm from the placental disc. The average thickness of the AM samples was about 100 μm. EDC, l -lysine, ninhydrin reagent, MMP-1 (EC 3.4.24.7), and MMP-9 (EC 3.4.24.35) were obtained from Sigma-Aldrich (St Louis, MO, USA). N-hydroxysuccinimide (NHS) was supplied by Acros Organics (Geel, Belgium). Balanced salt solution (BSS, pH 7.4) was purchased from Alcon Laboratories (Fort Worth, TX, USA). Dulbecco’s Modified Eagle’s Medium and TRIzol reagent were purchased from Gibco-BRL (Grand Island, NY, USA). The 24-well tissue culture polystyrene plates (Falcon 353047) and cell culture inserts (Falcon 3095) were purchased from Becton Dickinson Labware (Franklin Lakes, NJ, USA). All other chemicals were of reagent grade and were used as received without further purification.
8
Immunohistochemical Validation of Ovarian Endometriosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Five ovarian endometriotic cysts obtained from patients treated with dienogest (Dienogest group) and five ovarian endometriotic cysts obtained from the patients not treated with dienogest (Control group) were evaluated by IHC to validate the data from the GO and Ingenuity® pathway analyses. Of the genes extracted by the GO and pathway analyses, CSF1, MST1, MMP-1, and MMP-3 were selected for the IHC analysis. Deparaffinized and rehydrated tissue sections were incubated in the PT Link pre-treatment system (Dako Agilent, Santa Clara, CA) for antigen retrieval and then processed on Autostainer Link 48 (Dako Agilent) in accord with the manufacturer’s protocol. The following primary antibodies were used: CST1 (1:100 dilution, cat.# ab52864; Abcam, Boston, MA), MST1 (1:50 dilution, cat.# HPA024036; Sigma-Aldrich), MMP-1 (1:100 dilution, cat.# 52631; Abcam), and MMP-3 (1:100 dilution, cat.# 52915; Abcam). The staining intensity was scored as follows: 0, none of the cells stained positively; 1, weak staining; 2, moderate staining; and 3, strong staining. The results were analyzed with Student’s t-test to compare the staining intensity for each gene between the Dienogest and Control groups, and p-values < 0.05 were accepted as significant.
9
Quantification of Collagenases Using ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
10
Western Blot Analysis of pYSTAT3 and MMP-1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
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!