Dulbecco’s modified Eagle’s medium (DMEM)/F12 medium and fetal bovine serum (FBS) were purchased from Invitrogen Life Technologies (Carlsbad, CA, USA). Recombinant human PDGF-BB was purchased from ACROBiosystems, Inc. (Newark, DE, USA) and AS-IV was purchased from Tauto Biotech (Shanghai, China). Dimethyl sulfoxide (DMSO) and MTT were purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse anti-smoothelin, anti-α-smooth muscle actin (α-SMA), anti-desmin, anti-phospho-p38 mitogen-activated protein kinase (MAPK), anti-p38 MAPK, anti-matrix metalloproteinase (MMP)2, anti-MMP9 and anti-GAPDH antibodies, as well as a goat anti-mouse secondary antibody, were obtained from Abcam (Cambridge, UK).
Anti matrix metalloproteinase mmp 2
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-matrix metalloproteinase (MMP)2 is a laboratory product used for the detection and quantification of MMP2 protein. MMP2 is an enzyme involved in the breakdown of extracellular matrix components. This product can be used for research purposes to study the role of MMP2 in various biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti mmp9, by Abcam (3 mentions)
Anti gapdh, by Abcam (2 mentions)
Anti p38 mapk, by Abcam (1 mentions)
Mouse anti smoothelin, by Abcam (1 mentions)
Dulbecco s modified eagle s medium dmem f 12 medium, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse secondary antibody, by Abcam (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Chemiluminescence detection kit, by Cytiva (1 mentions)
Bca protein assay kit, by Bio-Rad (1 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Gapdh, by Abcam (1 mentions)
Anti wnt5a, by Abcam (1 mentions)
Anti vegf receptor 2, by Abcam (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Proteintech (1 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Chemidoc xrs system, by Bio-Rad (1 mentions)
4 protocols using anti matrix metalloproteinase mmp 2
1
PDGF-BB and AS-IV Effects on VSMCs
2
Quantifying Protein Levels via Western Blot
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Total protein was extracted from cells with RIPA lysis buffer (Thermo Fisher Scientific, Inc.). Total protein was quantified using a BCA protein assay kit (Bio-Rad Laboratories, Inc.). Equal amounts (30 µg) of protein samples were separated by SDS-PAGE on 10% gels and then electrophoretically transferred onto PVDF membranes (EMD Millipore). The PVDF membranes were incubated with primary antibodies overnight at 4°C after blocking with 10% non-fat milk for 1 h at room temperature. Subsequently, the PVDF membranes were incubated with HRP-conjugated secondary antibody (1:5,000; cat. no. ab150077; Abcam) at room temperature for 2 h. Protein signals were detected using a chemiluminescence detection kit (Amersham; Cytiva) and a semi-quantitative analysis was conducted using ImageJ software (version 1.8.0; National Institutes of Health). The following primary antibodies were purchased from Abcam: Anti-matrix metalloproteinase (MMP)2 (1:1,000; cat. no. ab92536), anti-MMP9 (1:1,000; cat. no. ab76003), anti-N-cadherin (1:1,000; cat. no. ab76011) and anti-GAPDH (dilution, 1:1,000; cat. no. ab181602) antibodies.
3
Protein Quantification and Analysis in Cultured Cells and Rat Bladder Tissues
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Cultured cells were lysed in 1 × SDS sample buffer. The rat bladder tissues were lysed using RIPA lysis buffer containing protease inhibitor cocktail (Roche, Shanghai, China). The supernatants were collected after centrifugation at 13,000 g at 4°C for 30 min. The protein concentration was quantified by a BCA protein assay kit (Beyotime, Nanjing, China). Equal amounts of protein were subject to 10% or 15% SDS-PAGE and transferred onto polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA), followed by blocking in 5% nonfat milk in distilled water for 2 h at room temperature. The membrane was then incubated overnight with the diluted primary antibody at 4°C, followed by incubation with horseradish-peroxidase-conjugated secondary antibody (Proteintech, Chicago, IL, USA) at room temperature for 2 h. The primary antibodies used were anti-matrix metalloproteinase (MMP) 2, anti-MMP9, anti-VEGF receptor 2, anti-Wnt5a, and anti-sFlt-1 (all obtained from Abcam). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a control (Abcam). The membranes with protein bands were analyzed using the ChemiDoc™ XRS system (Bio-Rad, Hercules, CA, USA). Densities of the bands of the Western blot images were evaluated using ImageJ software (National Institutes of Health).12 (link)
4
Protein Expression Analysis in Cells
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Protein was extracted from the cells with RIPA lysis buffer (Beyotime Institute of Biotechnology). Total protein was quantified using a BCA protein assay kit. Following extraction of the total protein from the cells, 10% SDS-PAGE was used to separate the proteins (30 µg), and then separated proteins were transferred to PVDF membranes. Membranes were blocked with 5% skimmed milk for 1 h at room temperature, and incubated with the following primary antibodies overnight at 4°C: Anti-Ki-67 (1:1,000; cat. no. ab245113; Abcam), anti-PCNA (1:1,000; cat. no. ab92552; Abcam), anti-matrix metalloproteinase (MMP)2 (1:1,000; cat. no. ab92536; Abcam), anti-MMP4 (1:1,000; cat. no. ab232865; Abcam),anti-MMP9 (1:1,000; cat. no. ab76003; Abcam) and anti-GAPDH (1:1,000; cat. no. ab8245; Abcam). Following which, membranes were incubated with a horseradish peroxidase-conjugated secondary antibody (1:5,000; cat. no. ab6721; Abcam) for 2 h at room temperature. Proteins were detected by the ECL™ Western Blotting Analysis System (GE Healthcare). The experiments were repeated at least 3 times. Protein expression levels were semi-quantified using ImageJ software (version 146; National Institutes of Health).
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