The following reagents were purchased from the indicated providers: dimethyl sulfoxide (DMSO; Sigma, D8418) and mifepristone (RU-486; Sigma, M8046). The following antibodies were used for immunoblotting: mouse anti-TurboGFP (Origene, TA150041), rabbit anti-TDP-43 (Proteintech, 10782-2-AP), rabbit anti-LC3 (MBL, PM036), mouse anti-Polyubiquitin (Enzo Life Science, BML-PW8805), mouse anti-Flag (Cell Signaling Technology, 2044), rabbit anti-HDAC6 (Santa Cruz Biotechnology, sc-11420), mouse anti-Lamin A/C (EMD Millipore, 05-714), HRP-conjugated anti-alpha-tubulin (Cell Signaling Technology, 9099), HRP-conjugated anti-rabbit IgG (Santa Cruz Biotechnology, sc-2004), HRP-conjugated mouse IgM (Abcam, ab97230), and HRP-conjugated mouse IgG (Santa Cruz Biotechnology, sc-2005). The following antibodies were used for immunocytochemistry (ICC): rabbit anti-cleaved caspase-3 (CC3) antibody (Cell Signaling Technology, 9664) and Alexa 594-conjugated anti-rabbit IgG (Jackson ImmunoResearch, 111-585-144). The following antibodies were used for immunohistochemistry: rat anti-ELAV (DSHB, RAT-ELAV-7), mouse anti-Polyubiquitin (Enzo Life Science, BML-PW8805), Alexa-488 conjugated rat IgG (Jackson ImmunoResearch, 112-545-167), and Alexa-594 conjugated mouse IgM (Jackson ImmunoResearch, 115-587-020).
Mouse anti turbogfp
Manufactured by OriGene
The Mouse anti-TurboGFP is a monoclonal antibody that specifically recognizes the TurboGFP protein, a variant of the green fluorescent protein (GFP) derived from the jellyfish Aequorea victoria. This antibody can be used to detect the presence and expression of TurboGFP in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Hrp conjugated anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Ru486, by Merck Group (1 mentions)
Mouse anti polyubiquitin, by Enzo Life Sciences (1 mentions)
Mouse anti lamin a c, by Merck Group (1 mentions)
Rabbit anti tdp 43, by Proteintech (1 mentions)
Mouse anti flag, by Cell Signaling Technology (1 mentions)
Alexa 594 conjugated anti rabbit igg, by Jackson ImmunoResearch (1 mentions)
Rabbit anti hdac6, by Santa Cruz Biotechnology (1 mentions)
Hrp conjugated anti alpha tubulin, by Cell Signaling Technology (1 mentions)
Hrp conjugated mouse igg, by Santa Cruz Biotechnology (1 mentions)
Permafluor mounting medium, by Thermo Fisher Scientific (1 mentions)
Lsm 700 confocal laser scanning microscope, by Zeiss (1 mentions)
Ta150041, by Merck Group (1 mentions)
Mouse anti gfp, by Merck Group (1 mentions)
Mouse anti ha, by Thermo Fisher Scientific (1 mentions)
Rabbit anti prrt2, by Merck Group (1 mentions)
3 protocols using mouse anti turbogfp
1
Protein Immunoblotting and Immunostaining
2
PLA Assay for Protein-Protein Interactions
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A standardized procedure for PLA was used (Jalili et al. 2018 (link)). Briefly, HEK293 cells were seeded on circular coverslips and transfected with specific plasmids for 72 h using a 3:1 ratio of PEI to pDNA. Cells were fixed with 4% paraformaldehyde for 10 min, followed by permeabilization and blocking (10% heat-inactivated goat serum, 1% Triton X-100) for 1 h. Cells were then probed with two specific primary antibodies raised in different species—mouse anti-TurboGFP (1:250; Origene TA150041)/rabbit anti-Flag (1:500; Sigma F7425)—overnight at 4°C. The cells were incubated with PLA probes for 1 h at 37°C, with ligase for 30 min at 37°C, and with polymerase for 100 min at 37°C, based on the manufacturer's protocols. Cells were stained with DAPI and then mounted with PermaFluor mounting medium (Thermo Scientific TA030FM). Images were captured on a Zeiss LSM 700 laser scanning confocal microscope.
3
Surface Labeling of PRRT2 Constructs
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Cells transfected with the various PRRT2 constructs were live-labeled by diluting primary antibodies (rabbit anti-PRRT2, 1:200, Sigma-Aldrich; mouse anti-HA, 1:200, Invitrogen; mouse anti-turboGFP, 1:200, Origene; and mouse anti-GFP, 1:500, Millipore) in culture medium for 1 h at 37 °C in a 5% CO2 incubator to detect the surface epitopes, followed by incubation with Alexa Fluor 488 or 594 secondary antibodies for 1 h at 37 °C, and fixed. In the experiments of live-labeling with HA antibodies, after live incubation with Alexa Fluor 488 secondary antibodies, cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100, and stained with anti-PRRT2 antibodies, followed by Alexa Fluor 594 secondary antibodies to label total PRRT2.
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