RIPA buffer (R0010; Beijing Solarbio) was used to prepare cell lysates. After the protein was quantified and denatured, the lysate (40 µg) was separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and transferred to a nitrocellulose membrane. The membranes were incubated with the following primary antibodies: anti‐OPN5 (GTX100173; GeneTex, Irvine, CA, USA), anti‐TYR (GTX16389, GeneTex), anti‐TRP1 (ab235446; Abcam), anti‐TRP2 (ab221144; Abcam), anti‐MITF (ab12039; Abcam), anti‐phospho‐MITF (ab59201; Abcam), anti‐PKC (AF6197; Affinity Biosciences, Cincinatti, OH, USA), anti‐phospho‐PKC (AF3197; Affinity), anti‐phospho‐CAMKII (MD1677‐100; MDL Medical Discovery Leader Biotech Co., Ltd, Beijing, China; mdlbiotech.com), anti‐CAMKII (MD2007‐100; MDL), anti‐actin (MD409‐020), anti‐p38 MAPK (MD2025‐100; MDL), anti‐phospho‐p38 MAPK (MD2084‐100; MDL), anti‐ERK1/2 (MD1853‐100), anti‐phospho‐ERK1/2 (MD1412‐100; MDL), anti‐JNK (MD1929‐20; MDL) and anti‐phospho‐JNK (MD1483‐20; MDL) for specific protein detection, and then incubated with horseradish peroxidase‐conjugated secondary antibodies. Specific bands were visualized using a chemiluminescent reaction (electrogenerated chemiluminescence) (E003‐50; 7Seabiotech, Shanghai, China; 7seapharmatech.com).
Gtx16389
Manufactured by GeneTex
Sourced in United States
GTX16389 is a laboratory equipment product. It is designed for scientific research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Af6197, by Affinity Biosciences (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
Ab221144, by Abcam (1 mentions)
Ab12039, by Abcam (1 mentions)
Triton x 100, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ab6310, by Abcam (1 mentions)
Alexa fluor 488 goat anti mouse secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
2 protocols using gtx16389
1
Western Blot Analysis of Melanogenic Proteins
2
Immunofluorescence Staining of Primary Melanocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human primary melanocytes were cultured on 24-well cell culture plates under standard conditions, as described above. The cells emigrating from the skin explants were fixed in 4% formaldehyde, permeabilized in 0.1% Triton X-100 (Sigma) and subsequently incubated for 30 min at room temperature in a blocking solution of 3% bovine serum albumin (BSA). Then, cells were incubated for 1 h at RT with rabbit anti-tyrosinase (1:100, GTX16389; GeneTex, CA, USA) and mouse anti-collagen III (1:200, ab6310; Abcam, Cambridge, MA, USA) antibodies. For the detection of the primary antibodies, Alexa Fluor 594 goat anti-rabbit and Alexa Fluor 488 goat anti-mouse secondary antibodies (1:200, Thermo Fisher Scientific Inc.) were used. The nuclei were counterstained with DAPI (Thermo Fisher Scientific Inc.).
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