α-Melanocyte-stimulating hormone (10 ng/mL; Sigma, c#M4135) was added to 0.5% human patient serum-supplemented endothelial medium to rescue the effect of MN serum on the GFB. The chip was incubated for 24 h. After 24 h, the serum-supplemented media was removed from the chips and the albumin assays was performed as described above.
α melanocyte stimulating hormone
Manufactured by Merck Group
Sourced in United States
α-melanocyte stimulating hormone is a lab equipment product. It is a peptide hormone that plays a role in the regulation of melanin production in melanocytes.
Automatically generated - may contain errors
Lab products found in correlation
Mushroom tyrosinase, by Merck Group (2 mentions)
Arbutin, by Merck Group (2 mentions)
Gsk3β, by Cell Signaling Technology (2 mentions)
α msh, by Merck Group (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
L 3 4 dihydroxyphenylalanine l dopa, by Merck Group (1 mentions)
L 4 hydroxyphenylalanine l tyrosine, by Merck Group (1 mentions)
3 isobutyl 1 methylxanthine, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trolox, by Merck Group (1 mentions)
Phenylmethylsulfonyl fluoride, by Merck Group (1 mentions)
Potassium dihydrogen phosphate, by Merck Group (1 mentions)
2 2 azino bis, by Merck Group (1 mentions)
Neuropeptide y, by Merck Group (1 mentions)
β endorphin, by Merck Group (1 mentions)
Phospho creb, by Cell Signaling Technology (1 mentions)
Phospho gsk3β, by Cell Signaling Technology (1 mentions)
Pd98059, by Merck Group (1 mentions)
7 protocols using α melanocyte stimulating hormone
1
Rescuing Endothelial Dysfunction with α-MSH
2
Tyrosinase Inhibitor Assay Protocol
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The kojic acid, trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), l -4-hydroxyphenylalanine (l -tyrosine), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), l -3,4-dihydroxyphenylalanine (l -dopa), phenylmethylsulfonyl fluoride (PMSF), α-melanocyte stimulating hormone (α-MSH), 3-isobutyl-1-methylxanthine (IBMX), 2,2-diphenyl-1-picrylhydrazyl (DPPH), 4-(1,1,3,3-tetramethylbutyl)phenylpolyethylene glycol (TritonTM X-100), dimethyl sulfoxide (DMSO), 3-morpholinosydnonimine (SIN-1), potassium hydrogen phosphate, potassium dihydrogen phosphate, and mushroom tyrosinase were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Hypothalamic Peptide Binding Assay
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The antiserum obtained (gC2 (1:30,000)) was incubated with 10 mmol of several hypothalamic peptides dissolved in 100 or 200 µl distilled water, or an equal volume of distilled water (control), for 1 h at RT and overnight at 4°C.
The pre-treated serum was then used for kisspeptin single-labeling immunohistochemistry as described above. The following peptides were used: rat- or human-type kisspeptin-10 and GnRH (Peptide Institute, Minoh, Osaka, Japan), NKB,
β-endorphin (ovine), Neuropeptide Y (NPY, ovine), α-melanocyte-stimulating hormone (Sigma-Aldrich, St Louis, MO, USA), Dyn (porcine), prolactin-releasing peptide-31 (bovine), prepro-RF-amide-related peptide (RFRP) (rat, 103-125),
pyroglutamylated-RF-amide peptide (rat, 13-26), and SP (human, 2-11) (Phoenix Pharmaceuticals Inc).
The anti-SP monoclonal antibody (1:2,000,000) was incubated with either SP (10 nmol), NKB (10 nmol), or 100 µl distilled water for 1 h at RT and overnight at 4°C, and subjected to SP immunohistochemistry using a similar protocol
as the kisspeptin single-labeling immunohistochemistry, except that the normal serum and the second antibody were substituted with normal horse serum and anti-mouse IgG, respectively.
The pre-treated serum was then used for kisspeptin single-labeling immunohistochemistry as described above. The following peptides were used: rat- or human-type kisspeptin-10 and GnRH (Peptide Institute, Minoh, Osaka, Japan), NKB,
β-endorphin (ovine), Neuropeptide Y (NPY, ovine), α-melanocyte-stimulating hormone (Sigma-Aldrich, St Louis, MO, USA), Dyn (porcine), prolactin-releasing peptide-31 (bovine), prepro-RF-amide-related peptide (RFRP) (rat, 103-125),
pyroglutamylated-RF-amide peptide (rat, 13-26), and SP (human, 2-11) (Phoenix Pharmaceuticals Inc).
The anti-SP monoclonal antibody (1:2,000,000) was incubated with either SP (10 nmol), NKB (10 nmol), or 100 µl distilled water for 1 h at RT and overnight at 4°C, and subjected to SP immunohistochemistry using a similar protocol
as the kisspeptin single-labeling immunohistochemistry, except that the normal serum and the second antibody were substituted with normal horse serum and anti-mouse IgG, respectively.
4
Melanin Synthesis Mechanism Investigation
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Dulbecco’s modified Eagle’s medium (DMEM) was purchased from WElGENE Inc. (Republic of Korea). Gallic acid was purchased from Sigma-Aldrich (St. Louis, MO, USA) for use in high-performance liquid chromatography (HPLC) and for investigating the mechanism of Gallic acid on melanin synthesis. α-melanocyte-stimulating hormone, arbutin, LY294002, and PD98059 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies recognizing GAPDH, AKT, phospho-AKT, GSK-3β, phospho-GSK-3β, CREB, phospho-CREB, PKA, and phospho-PKA were obtained from Cell Signaling Technology (Beverly, MA, USA). Other primary antibodies and secondary antibodies were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). All other unlabeled chemicals and reagents were analytical grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).
5
Melanogenesis Pathway Modulation Protocol
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Sigma-Aldrich (St. Louis, MO, USA) provided the 6-methylcoumarin (CAS 92-48-8), 7-methylcoumarin (CAS 2445-83-2), 4-hydroxy-6-methylcoumarin (CAS 13252-83-0), 4-hydroxy-7-methylcoumarin (CAS 18692-77-8), α-melanocyte-stimulating hormone (α-MSH), a protease/phosphatase inhibitor cocktail, sodium hydroxide (NaOH), and L-DOPA. Biosesang (Seongnam, Gyeonggi-do, Korea) supplied the MTT, DMSO, PBS, TBS, SDS, a RIPA buffer, and an ECL kit. Thermo Fisher Scientific (Waltham, MA, USA) provided the DMEM, penicillin–streptomycin, a BCA protein assay kit, and 0.5% trypsin-ethylenediaminetetraacetic acid (10×). The primary antibodies, tyrosinase, TRP-1, TRP-2, and MITF, were purchased from Santa Cruz Biotechnology (Dallas, TX, USA), and the other antibodies, 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, and antirabbit, and secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Skim milk was purchased from BD Difco (Sparks, MD, USA), the fetal bovine serum (FBS) was purchased from Merck Millipore (Burling, VT, USA), and the Tween 20 and 2× Laemmle sample buffer were obtained from Bio-rad (Hercules, CA, USA).
6
Evaluating Melanogenesis Inhibitors in Dermal and Skin Cell Lines
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Neonatal, primary HDF cells (a human dermal fibroblast cell line) were obtained from Lifeline (Oceanside, CA, USA). HaCaT (a human keratinocyte cell line), HEK293 (a human embryonic kidney 293 cell line), and B16F10 cells (a murine melanoma cell line) were purchased from the American Type Culture Collection (Rockville, MD, USA). Dulbecco’s modified Eagle’s medium (DMEM), penicillin, streptomycin, phosphate-buffered saline (PBS), and fetal bovine serum (FBS) were purchased from HyClone (Grand Island, NY, USA). Mushroom tyrosinase, polyethylenimine (PEI), H2DCF-DA, Kojic acid, arbutin, SB203580, SP600125, α-melanocyte stimulating hormone, U0126, BAY11-7082, and Annexin V-FITC Apoptosis Detection Kit were obtained from Sigma-Aldrich (St. Louis, MO, USA). Specific antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA) or Santa Cruz Biotechnology (Santa Cruz, CA, USA).
7
Melanin Extraction from SK-MEL2 Cells
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Two hundred nM of α-melanocyte stimulating hormone (α-MSH; Sigma-Aldrich, USA) and test samples were treated in http://www.e-ajbc.org cultured SK-MEL2 and cultured for 48 h at 37℃ under 5% CO2.
Cells were harvested using trypsin-EDTA and centrifuged at 12000 rpm for 10 min. One molar NaOH was added to dissolve melanin on cell pellets.
Cells were harvested using trypsin-EDTA and centrifuged at 12000 rpm for 10 min. One molar NaOH was added to dissolve melanin on cell pellets.
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