Medium 254 and human melanocyte growth supplement were purchased from Cascade Biologics (Portland, OR). Dulbecco modified Eagle medium, penicillin‐streptomycin and 0.5% trypsin‐ethylene diamine tetra acetic acid were obtained from GIBCO Invitrogen (Auckland, NZ). The primary and secondary antibodies used in this study were purchased from Santa Cruz Biotech (Santa Cruz, CA), Cell Signaling Technology (Danver, MA), or BD Biosciences (San Jose, CA). Chemiluminescence detection kits were purchased from GE Healthcare (Piscataway, NJ). All other chemicals, including α‐MSH, were purchased from Sigma‐Aldrich (St Louis, MO).
Protocol detail
Melanocyte Growth Supplement Protocol
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Medium 254
human melanocyte growth supplement
Dulbecco modified Eagle medium
penicillin‐streptomycin
Chemiluminescence detection kits
α‐MSH
Acetic acid
Antibodies
Biologics
Chemiluminescence
Eagle
Ethylene diamine
Human
Melanocyte
Penicillin
Streptomycin
Supplement
Tetra
Trypsin
α msh
Corresponding Organization : Korea University
Other organizations : Korea Institute of Oriental Medicine
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Variable analysis
independent variables
- Medium 254
- Human melanocyte growth supplement
- Not explicitly mentioned
- Dulbecco modified Eagle medium
- Penicillin‐streptomycin
- 0.5% trypsin‐ethylene diamine tetra acetic acid
- Primary and secondary antibodies
- Chemiluminescence detection kits
- α‐MSH
- Positive controls: Not specified
- Negative controls: Not specified
Annotations
Based on most similar protocols
The protocols mention the use of various chemical reagents, including α-MSH, DMSO, MbBr, PMSF, pyrogallol, NBT, NADPH, H2O2, BSA, SP600125, PD98059, L-DOPA, minocycline hydrochloride, DCFH2-DA, penicillin, amphotericin B, and phosphatase inhibitor cocktail. These reagents are critical for the experiments and should be of high quality and properly stored to ensure reproducibility (Protocols 1, 2, 3).
The protocols indicate the use of various cell lines, including primary human melanocytes, the PIG1 melanocyte cell line, the MNT-1 malignant melanoma cell line, and the B16F10 cell line. The use of appropriate cell lines and their proper maintenance conditions (e.g., temperature, CO2 levels) are crucial for the success of the experiments (Protocols 4, 5).
The protocols mention the use of various growth media, such as DMEM, M-254, and M-154, as well as growth supplements like HMGS and HKGS. The appropriate selection and preparation of culture media are important for maintaining the viability and functionality of the cells (Protocols 1, 2, 3, 4, 5).
The protocols highlight the use of various antibodies, including tyrosinase, β-actin, phospho-JNK, phospho-p38, and phospho-ERK antibodies, for Western blotting or immunofluorescence experiments. The proper selection, dilution, and validation of these antibodies are crucial for accurate and reliable results (Protocols 1, 2, 4).
The protocols mention the use of various detection kits, such as ECL kits and the Cell Proliferation Reagent WST-1, for visualizing and quantifying the experimental results. The appropriate use and validation of these detection methods are important for ensuring the accuracy and reproducibility of the experiments (Protocols 1, 2, 3).
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