Total cellular proteins were extracted by MSCs using lysis buffer (0.1 M KH2PO4, pH 7.5, 1% NP-40, 0.1 mM β-glycerolphosphate, supplemented with protease inhibitor cocktail; Sigma-Aldrich) and quantified spectrophometrically by the Bio-Rad Protein Assay (Bio-Rad Laboratories). Thirty micrograms of proteins were subjected to 8–12% SDS-PAGE and transferred to nitrocellulose membrane (GE Healthcare Life Sciences) at 30 mA for 2 hours and 30 min. Membranes were blocked with 5% non-fat dry milk in TBS-Tween for 1 hour at room temperature and incubated with primary antibodies against OPN (1:500; Millipore), BMP-2 (1:1000; Santa Cruz), PPAR-γ (1:200; Santa Cruz) and β-actin (clone AC-74; Sigma-Aldrich) at 4 °C o/n. Incubation with secondary antibody human anti-rabbit/mouse horseradish peroxidase-conjugated (GE Healthcare) was performed for 1 hour at room temperature. The protein signal was detected using Westar ηC chemiluminescent substrate (Cyanagen) and band intensities were quantified by densitometry analysis by ImageJ software (NIH, USA). Similarly, equal volumes (18 μl) of MSC supernatants, previously cultured in serum-free DMEM for 24 hours and stored at −80 °C, were loaded into 8% SDS-PAGE and transferred to nitrocellulose membrane as already described, using primary MMP-9 antibody (1:1000; Cell Signaling).
Mmp 9 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The MMP-9 antibody is a laboratory tool used to detect and measure the presence of matrix metalloproteinase-9 (MMP-9) in biological samples. MMP-9 is an enzyme involved in the breakdown of extracellular matrix proteins, which is a crucial process in various physiological and pathological conditions. The MMP-9 antibody can be used in techniques such as Western blotting, ELISA, and immunohistochemistry to quantify or visualize MMP-9 levels in research applications.
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Lab products found in correlation
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Nitrocellulose membrane, by Cell Signaling Technology (1 mentions)
β actin clone ac 74, by Merck Group (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Protein assay, by Bio-Rad (1 mentions)
Bmp 2, by Santa Cruz Biotechnology (1 mentions)
Westar ηc chemiluminescent substrate, by Cyanagen (1 mentions)
Pparγ, by Santa Cruz Biotechnology (1 mentions)
N cadherin antibody, by Cell Signaling Technology (1 mentions)
Vimentin antibody, by Cell Signaling Technology (1 mentions)
β actin, by Merck Group (1 mentions)
Snail antibody, by Cell Signaling Technology (1 mentions)
V5 antibody, by Merck Group (1 mentions)
Phospho stat5, by Cell Signaling Technology (1 mentions)
Jak1 antibody, by Cell Signaling Technology (1 mentions)
β actin antibody, by Cell Signaling Technology (1 mentions)
Phospho stat6, by Cell Signaling Technology (1 mentions)
Pkcζ antibody, by Cell Signaling Technology (1 mentions)
Plcγ1 antibody, by Cell Signaling Technology (1 mentions)
Phospho stat2, by Cell Signaling Technology (1 mentions)
8 protocols using mmp 9 antibody
1
Protein Extraction and Analysis of MSCs
2
Western Blot Antibody Profiling
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Antibodies against Nudt2 (#10484-1-AP, dilution 1:1000, Proteintech, Biotest, Kfar Saba, Israel), β-Actin (#A1978, dilution 1:10,000, Sigma-Aldrich, Jerusalem, Israel), V5-antibody (#v8012, dilution 1:2000, Sigma Aldrich), vimentin antibody (#5741T, dilution 1:2000, Cell Signaling, Jerusalem, Israel), N cadherin antibody (#13116, dilution 1:2000, Cell Signaling, Jerusalem, Israel), MMP9 antibody (#13667, dilution 1:2000, Cell Signaling, Jerusalem, Israel), and Snail antibody (#3879, dilution 1:2000, Cell Signaling, Jerusalem, Israel) were used.
3
Protein Kinase Signaling Pathway Analysis
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The following antibodies were purchased from Cell Signaling (Danvers, MA, USA): JAK1 Antibody (#3332, 130kDa), Phospho-JAK2 (Tyr1007/1008) Antibody (#3771, 125kDa), β-Actin Antibody (#4967, 45kDa), Phospho-STAT1 (Tyr701) (58D6) Rabbit mAb (#9167, 84,91kDa), Phospho-STAT2 (Tyr690) Antibody (#4441, 113kDa), Phospho-STAT3 (Tyr705) Antibody (#9131, 79,86kDa), Phospho-STAT5 (Tyr694) Antibody (#9351, 90kDa), Phospho-STAT6 (Tyr641) Antibody (#9361, 110kDa), Ras Antibody (#3965, 21kDa), p44/42-MAPK (Erk1/2) Antibody (#9102, 42,44kDa), GSK-3β (27C10) Rabbit mAb (#9315, 46kDa), MMP-9 Antibody (#3852, 84,92kDa), Phospho-PKCα/β II (Thr638/641) Antibody (#9375, 80,82kDa); PKCα Antibody (#2056, 80kDa); Phospho-PKC (pan) (βII Ser660) Antibody (#9371, 78 a 85kDa); Phospho-PKD/PKCμ (Ser744/748) Antibody (#2054, 115kDa); PKD/PKCμ Antibody (#2052, 115kDa); PKCδ Antibody (#2058, 78kDa); Phospho-PKCδ/θ (Ser643/676) Antibody (#9376, 78kDa), PKCζ Antibody (#9372, 78kDa); PLCγ1 Antibody (#2822, 155kDa); anti-mouse, anti-rabbit and anti-goat IgGs antibodies. From Abcam (Cambridge, MA, USA): anti-PKC antibody (ab59363) was purchased. PepChip1-Kinomics™ slides were obtained from Pepscan Presto BV (Lelystad, the Netherlands).
WP1066 (#573097; C17H14BrN3O) and Tofacitinib (#PZ0017; C16H20N6O · C6H8O7) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
WP1066 (#573097; C17H14BrN3O) and Tofacitinib (#PZ0017; C16H20N6O · C6H8O7) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
4
Molecular Mechanisms of Sulforaphane-Induced Apoptosis
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Culture media were purchased from Hyclone (Logan, Utah, USA), FBS and penicillin–streptomycin were acquired from Invitrogen (Carlsbad, CA, USA). MMP-9 antibody was purchased from Cell Signaling Technology, Inc (Shanghai, China). MMP-2 antibody was supplied by Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against Bax, Bad, Bcl-2, cytochrome C and α-tubulin were purchased from Proteintech Group, Inc (Chicago, USA). d , l -Sulforaphane (SFN) was acquired from Sigma (St Louis, MO, USA). MTS assay kit was purchased from Promega (Madison, USA). Transwell plates for invasion assay were bought from BD Biosciences (Bedford, MA, USA). BCA protein assay kit was obtained from Thermo Scientific Pierce Protein Research Products (Rockford, IL., USA). Annexin V-FITC Apoptosis Assay Kit was acquired from Nanjing KeyGEN Biotech (Nanjing, China).
5
Platelet Signaling Pathway Analysis
6
Investigating Therapeutic Targets in Myeloma
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T3, T4 and tetrac (Sigma-Aldrich, St. Louis, MO, USA) were dissolved to 1 mM in KOH-PG (final concentration of 0.04 N KOH with 0.4% polyethylene glycol (vol/vol). RGD/RGE peptides (Sigma) were dissolved to 100mM in PBS). Bortezomib was obtained from the oncology pharmacy at Meir Medical Center. Vehicle control was used in each experiment. APC-CD138 antibodies (clone B-B4) were from Miltenyi Biotec, Bergisch Gladbach, Germany. αvβ3 (LM609 unconjugated/PE) monoclonal antibody was from Chemicon International, Harrow, UK. U0126 was purchased from Cell Signaling Technology, Danvers, MA, USA and MMP-9 antibody (#3852) was obtained from Cell Signaling (Boston, MA, USA).
7
Evaluation of Neuroinflammation Markers
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The ipsilateral hemisphere was collected 24 hours after reperfusion and processed for a Western blot analysis as described previously. 14 The first antibodies were an anti-myeloperoxidase (MPO) antibody (Cell Signaling Technology, Danvers, MA, USA, dilution 1:2000) as a marker for neutrophils, an anti-matrix metalloproteinase (MMP)-9 antibody (Cell Signaling Technology, Danvers, MA, USA, dilution 1:2000), and anti-intercellular molecule-1 (ICAM-1) antibody (Abcam, Cambridge, MA, USA, dilution 1:2000). Western blot bands were quantified by densitometry using an image analysis system (ImageJ 1.37v; NIH Bethesda, MD, USA)
8
Molecular Profiling of Ocular Angiogenesis
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Fresh isolated eye tissues and cells were lysed, quantified, blotted and developed as previously described. 18 The following primary antibodies were used for the purpose of this experiment: goat anti-mouse endocan antibody (0.1 lg/mL; R&D Systems); rabbit anti-matrix metalloproteinase (MMP)-2 antibody (1:1000; Cell Signaling Technology); MMP-9 antibody (1:1000; Cell Signaling Technology); rabbit anti-PIGF antibody (1:1000; Abcam, Cambridge, UK); goat anti-VEGF antibody (1:1000; Abcam); rabbit anti-VEGFR1 antibody (1:1000; Abcam); rabbit anti-VEGFR2 antibody (Cell Signaling Technology); rabbit anti-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-phospho (p)-P38 antibody (1:1000; Cell Signaling Technology); rabbit anti-extracellular signal-regulated kinase (ERK) antibody (1:1000; Cell Signaling Technology); rabbit anti-p-ERK (1:1000; Cell Signaling Technology); rabbit anti-c-Jun Nterminal kinases (JNK) antibody (1:1000; Cell Signaling Technology) and rabbit anti-p-JNK antibody (1:1000; Cell Signaling
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