Syringetin (CAS 4423-37-4) was purchased from Extrasynthese (Genay Cedex, France), while α-melanocyte stimulating hormone (α-MSH), protease/phosphatase inhibitor cocktail, sodium hydroxide (NaOH), and L-DOPA were obtained from Sigma-Aldrich (St. Louis, MO, USA). MTT, DMSO, PBS, TBS, SDS, RIPA buffer, and the ECL kit were purchased from Biosesang (Seongnam, Gyeonggi-do, Korea), while DMEM, penicillin–streptomycin, BCA protein assay kits, and 0.5% trypsin-ethylenediaminetetraacetic acid (10×) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). For Western blots, the primary antibodies against tyrosinase, TRP-1, TRP-2, and MITF were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), while the other antibodies, against p-ERK, ERK, p-p38, p38, p-JNK, JNK, p-PKA, PKA, p-AKT, AKT, p-GSK-3β, GSK-3β, p-β-catenin, β-catenin, β-actin, anti-rabbit, and secondary antibodies, were purchased from Cell Signaling Technology (Danvers, MA, USA). Skimmed milk was purchased from BD Difco (Sparks, MD, USA), fetal bovine serum (FBS) from Merck Millipore (Burling, USA), and Tween 20 and 2 × Laemmli sample buffer were obtained from Bio-rad (Hercules, CA, USA).
Ecl kit
Manufactured by Biosesang
The ECL kit is a laboratory equipment used for the detection and quantification of proteins in Western blot analysis. It contains the necessary reagents for the enhanced chemiluminescent (ECL) detection of proteins that have been labeled with a horseradish peroxidase (HRP)-conjugated antibody.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Laemmli sample buffer, by Bio-Rad (1 mentions)
L dopa, by Merck Group (1 mentions)
Tween 20, by Bio-Rad (1 mentions)
Phosphate buffered saline (pbs), by Biosesang (1 mentions)
Dimethyl sulfoxide (dmso), by Biosesang (1 mentions)
P gsk3β, by Cell Signaling Technology (1 mentions)
α melanocyte stimulating hormone α msh, by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Biosesang (1 mentions)
Sodium dodecyl sulfate (sds), by Biosesang (1 mentions)
Tris buffered saline (tbs), by Biosesang (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Protease phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
2 protocols using ecl kit
1
Melanogenesis Regulation via Signaling Pathways
2
Keratinocyte Response to P. acnes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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HaCaT keratinocytic cells were plated at 1 × 10 5 cells/well in 12-well plates and cultured overnight. The cells were then pretreated with CC extracts or berberine for 1 h. After stimulating the cells with P. acnes for 24 h, total cellular proteins were prepared using PBS lysis buffer. These proteins were separated using 10% SDS-PAGE and transferred to PVDF membranes. These membranes were then blocked and incubated with a primary antibody overnight in a shaker at 4°C. After washing three times with TBST, the membranes were incubated for 1 h with a horseradish peroxidase-conjugated secondary antibody. Protein bands were detected using an ECL kit (Biosesang, Korea) and ChemiDoc system (Bio-Rad, USA). All experiments were repeated three times and the most obvious results are presented.
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