Cell lines were transfected with LTX lipofectamine (Thermo Fisher, A12621) following manufacturer's instructions with pLAralar alone or together with pCMV5-AlbEnh-βglobin-mCitrin-Flag (pLCitrin-Flag) generated similarly to pLAralar from slc25a13/AGC2/citrin cDNA ORF Clone (Sino Biological). Fixed cells in 4% PFA were blocked (10% Horse serum, PBS) and probed with polyclonal antibody raised against aralar (1/500; [3 (link)]) and either monoclonal antibody against Flag epitope (Sigma, F1804, 1/500) or monoclonal antibody against citrate synthase (Santa Cruz, sc-390693, 1/500) and appropiate secondary antibodies. To visualize nuclei, DAPI (1 μg/ml; Merck) was used. Images were obtained at Axiovert 200 M inverted microscope equipped with a 100×/1.3 Plan-Neofluor objective with appropriate filters.
Sc 390693
Manufactured by Santa Cruz Biotechnology
Sc-390693 is a lab equipment product manufactured by Santa Cruz Biotechnology. It is designed for use in various scientific research applications. The core function of this product is to facilitate specific experimental procedures, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Orf clone, by Sino Biological (1 mentions)
Lipofectamine ltx, by Thermo Fisher Scientific (1 mentions)
F1804, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Imp1 2 3, by Santa Cruz Biotechnology (1 mentions)
Igf2bp2, by Proteintech (1 mentions)
Protease inhibitor cocktails 1 and 2, by Merck Group (1 mentions)
2 protocols using sc 390693
1
Visualizing Aralar and Citrin Protein Localization
2
Protein Extraction and Western Blotting
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Protein was extracted with RIPA buffer supplemented with protease inhibitor cocktails I and II (Sigma) and phosphatase inhibitor cocktail set III (Calbiochem). Protein was quantified with Pierce BCA protein assay kit (Thermo Fisher) and analysed with a Sunrise Tecan plate reader. After SDS-PAGE and electro-transfer onto PVDF membranes, Western blotting was performed with the following primary antibodies and concentrations: Lin28a (1:1000, CST), Pax7 (0.28μg/ml, DSHB), Pax3 (0.31μg/ml, DSHB), MyoD (1:1000, Santa Cruz Biotechnology), MHC (0.23μg/ml, DSHB), MyoG (1:1000, Santa Cruz Biotechnology), IGF2BP2 (1:1000, Proteintech), Hmga2 (1:1000, CST), Cre (1:1000, Millipore), GAPDH (1:1000, CST), Vinculin (K106900P, 1:5000, Solarbio), Tubulin (ab210797; Abcam; 1:1000), P53 (sc-126; Santa Cruz; 1:100), IMP1/2/3 (sc-271785; Santa Cruz; 1:1000), citrate synthase (G-3) (sc-390693; Santa Cruz ; 1:1000).
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