Recombinant human MG53 (GST-tagged) was purchased from Cyclex (Japan, # CY-R2073). The plasmid encoding GFP-tagged TRIM72 (GFP-MG53) and the corresponding empty vector (GFP-Turbo) were purchased from OriGene Technologies, Inc. (Rockville, USA). Insulin was provided by Sigma Aldrich (St Quentin Fallavier, France).
Turbo gfp
Manufactured by OriGene
Sourced in United States
Turbo-GFP is a fluorescent protein derived from the green fluorescent protein (GFP) of the jellyfish Aequorea victoria. It is engineered to have enhanced fluorescence properties, making it a useful tool for various biological applications.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (3 mentions)
Gfp mg53, by OriGene (2 mentions)
Insulin, by Merck Group (2 mentions)
Hsp90, by Cell Signaling Technology (1 mentions)
Vinculin, by Abcam (1 mentions)
Total akt, by Cell Signaling Technology (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
Peroxidase conjugated anti mouse and anti rabbit iggs, by Thermo Fisher Scientific (1 mentions)
Ndufs1, by GeneTex (1 mentions)
Cox5b, by Santa Cruz Biotechnology (1 mentions)
Ecl prime western blotting detection reagent, by Cytiva (1 mentions)
Chemidoc mp imager, by Bio-Rad (1 mentions)
β actin, by Merck Group (1 mentions)
Image lab, by Bio-Rad (1 mentions)
Ndufb8, by Abcam (1 mentions)
Core 2, by Abcam (1 mentions)
Cox5a, by Abcam (1 mentions)
Alexa488 coupled goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Cf633nm coupled donkey anti rat, by Biotium (1 mentions)
E cadherin, by BD (1 mentions)
5 protocols using turbo gfp
1
Recombinant MG53 and TRIM72 Protocols
2
Recombinant MG53 Protein Purification
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Recombinant human MG53 (GST-tagged) was purchased from Cyclex (Japan, # CY-R2073).
The plasmid encoding GFP-tagged TRIM72 (GFP-MG53) and the corresponding empty vector (GFP-Turbo) were purchased from OriGene Technologies, Inc. (Rockville, USA). Insulin was provided by Sigma Aldrich (St QuentinFallavier, France).
The plasmid encoding GFP-tagged TRIM72 (GFP-MG53) and the corresponding empty vector (GFP-Turbo) were purchased from OriGene Technologies, Inc. (Rockville, USA). Insulin was provided by Sigma Aldrich (St QuentinFallavier, France).
3
Western Blot Analysis of Signaling Proteins
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Protein extracts (30μg) were fractionated on 10% polyacrylamide gel under reducing conditions (sample buffer containing 10 mM dithiothreitol (DTT)) and transferred onto nitrocellulose membranes. Membranes were blocked with 5% milk or BSA 5% in TBS-T 0.1% for 1h. Membranes were then washed three times with TBS-T 0.1% for 10 minutes and incubated overnight at 4°C in blocking buffer containing primary antibodies. Primary antibodies used: phospho-Akt Ser473 (#4060), total Akt (#4691), Hsp90 (# 4877) (Cell Signaling Technology, Danvers, MA), MG53 (Sigma Aldrich, # SAB2108735), Vinculin (Abcam, #ab73412) and Turbo-GFP (OriGene, # TA150041). After incubation with a secondary peroxidase-conjugated antibody, signals were detected by chemiluminescence (GE Healthcare).
4
Comprehensive Western Blot Analysis
5
Immunostaining Protocol for Thoracic Mammary Glands
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Thoracic mammary glands were fixed for 24 hours in 4% paraformaldehyde and embedded in paraffin wax. Paraffin sections of 5 µm were prepared and subjected to 1 mM disodium-EDTA antigen retrieval as described previously [28] (link). Primary antibodies used for immunofluorescence are the following: Cytokeratin 8 (Developmental Studies Hybridoma Bank TROMA-I, rat, 1∶100), Estrogen receptor (Novocastra NCL-ER-6F11, mouse, 1∶100), E-Cadherin (BD Biosciences 610181, mouse, 1∶250), Progesterone receptor (Abnova MAB9785, rabbit, 1∶400), Smooth muscle actin (Sigma A2547, mouse, 1∶1000), Tbx3 (Invitrogen 424800, rabbit, 1∶100), turboGFP (Pierce Antibodies PA522688, rabbit, 1∶400), turboGFP (OriGene TA150041, mouse 1∶250). Secondary antibodies used at 1∶400 dilution are from Invitrogen: Alexa488-coupled goat anti-mouse (A11029), Alexa488-coupled goat anti-rabbit (A11034), Alexa568-coupled goat anti-mouse (A11031) and Alexa568-coupled goat anti-rabbit (A11036). Additionally, CF633nm-coupled donkey anti-rat (Biotium 20137-1) was used at 1∶400 dilution. Images were acquired on a Zeiss LSM-710 confocal microscope with a pinhole aperture of 1 Airy unit.
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