Proteins extracted from CRC fresh tissues or cells with various treatments were fractionated by SDS-PAGE, transferred to polyvinylidene difluoride (PVDF) membrane, and then incubated with a primary specific antibody for ESCO2 (1:1,000, ab86003; Abcam, Cambridge, MA, USA), E-cadherin (1:1,000, sc-14472; Santa Cruz Biotechnology Inc.), N-cadherin (1:1,000, #13116; Cell Signaling Technology, Danvers, MA, USA), Vimentin (1:1,000, #5741; Cell Signaling Technology), MMP2 (1:1,000, #87809; Cell Signaling Technology), active MMP2 (1:500; KeyGen, Jiangsu, China), and β-actin (1:1,000, #4970; Cell Signaling Technology) in 5% of nonfat milk, followed by a horseradish peroxidase (HRP)-conjugated anti-rabbit/mouse secondary antibody. Enhanced chemiluminescence (ECL) detection reagent (Amersham Life Science, Piscataway, NJ, USA) was used to visualize the results.
Enhanced chemiluminescence ecl detection reagent
Manufactured by Cytiva
Sourced in United States
The Enhanced chemiluminescence (ECL) detection reagent is a laboratory product designed to facilitate the detection of proteins in Western blot analysis. It provides a reliable and sensitive method for visualizing target proteins labeled with specific antibodies.
Automatically generated - may contain errors
Lab products found in correlation
β actin, by Cell Signaling Technology (1 mentions)
Mmp 2, by Cell Signaling Technology (1 mentions)
E cadherin, by Santa Cruz Biotechnology (1 mentions)
N cadherin, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
Zymogram gel, by Thermo Fisher Scientific (1 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (1 mentions)
Aprotinin, by Roche (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Chromozym try, by Roche (1 mentions)
Opti mem, by Merck Group (1 mentions)
Hybond p polyvinylidene difluoride pvdf membrane, by Cytiva (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Dulbecco modified eagle medium dmem, by Merck Group (1 mentions)
Horseradish peroxidase conjugated donkey anti rabbit immunoglobulin g igg, by Cytiva (1 mentions)
Fetal bovine serum 3 4 5 dimethyl thiazol 2 yl 2 5 diphenyl tetrazolium bromide mtt, by Merck Group (1 mentions)
Sheep anti mouse igg, by Cytiva (1 mentions)
Anti phospho pp2a c mab, by Santa Cruz Biotechnology (1 mentions)
4 protocols using enhanced chemiluminescence ecl detection reagent
1
Immunoblotting Analysis of CRC Proteins
2
Sulfur Mustard Impacts on Keratinocyte Proteases
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Normal human epidermal keratinocytes (NHEK), human keratinocyte growth supplement (Insulin, BPE, Transferrin, EGF, Hydrocortisone, PSA), and basal media for epithelial cells—Epilife with calcium, kit for splitting cells were purchased from Cascade Biologics (Portland, OR). Sulfur mustard (>98% pure) was from the Edgewood Chemical and Biological Center (ECBC), Aberdeen Proving Ground, Maryland. Protein molecular weight markers, zymogram gel, zymogram renaturing and developing buffers, precast SDS-PAGE gels and buffers were from Invitrogen (Carlsbad, CA). Anti-laminin-5 γ2 polyclonal antibody and horseradish peroxidase-conjugated secondary goat antibody were from Santa Cruz (Santa Cruz, CA). Enhanced chemiluminescence (ECL) detection reagent was from Amersham (Piscataway, CA). Protease inhibitor cocktail, chromozym TRY (serine specific substrate), and aprotinin were from Roche (Basel, Switzerland). Amastatin was purchased from Axxora, LLC (San Diego, CA). Dimethyl sulfoxide (DMSO), coomassie blue R-250, and E-64 were from Sigma (St. Louis, MO). Anti-membrane type matrix metalloptotease-1 antibody (MT-MMP-1), anti-matrix metalloprotease-2 antibody (MMP-2), and anti-matrix metalloprotease-9 (MMP-9) antibody were from Research Diagnostics, Inc. (Flanders, NJ).
3
Molecular Signaling Pathway Analysis
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The Dulbecco modified Eagle medium (DMEM), optiMEM, trypsin (0.25%), l -glutamine, penicillin, streptomycin, fetal bovine serum (FBS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Okadaic acid was obtained from Calbiochem (San Diego, CA, USA), and 3-O-Methyl-sphingomyeline (3-OME-SM) was purchased from Biomol (Plymouth Meeting, PA, USA). The anti-phospho-p44/p42 extracellular signal-regulated kinase (ERK) (Thr202/Tyr204) was purchased from Cell Signaling (Beverly, MA, USA). The anti-phospho-Akt (Ser473) and anti-Akt monoclonal antibodies (mAbs) were purchased from Biovision (Mountain View, CA, USA). The anti-phospho-PP2A-C mAb, and anti-demethylated-PP2A-C mAb were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The anti-α-tubulin mAb was purchased from Neo Markers (Fremont, CA, USA). The horseradish peroxidase conjugated donkey anti-rabbit immunoglobulin G (IgG) and sheep anti-mouse IgG were purchased from Amersham (Buckinghamshire, UK). The Hybond-P polyvinylidene difluoride (PVDF) membrane and enhanced chemiluminescence (ECL) detection reagent were purchased from Amersham. Ketamine was dissolved in 0.1% DMSO, and stored at 4 °C.
4
Western Blot Analysis of DDR-2 Protein
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Bicinchoninic acid (BCA) protein assay kit was purchased from Pierce (Bologna, Italy). Electrophoresis reagents were prepared from Bio-Rad (Hercules, CA, USA). Glycine, sodium dodecyl sulfate, Tris, polyacrylamide gel electrophoresis (PAGE), N-methylene-bisacrylamideand mercaptoethanol were purchased from Merk Co. (Germany). Rabbit polyclonal antibody against DDR-2 was purchased from Santa Cruz (Santa Cruz Biotech, Santa Cruz, CA, U.S.A.). Enhanced chemiluminescence (ECL) detection reagent was procured from Amersham (Amersham Corp., Arlington Heights, IL, USA). RNeasy Minikit was supplied by Qiagen (Qiagen Inc, USA). DNase was purchased from Fermentas (Burlington, Ontario, Canada). BCA protein assay kit was obtained from Pierce (TemaRicerca SrI, Bologna, Italy).
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