Sc 1657

Manufactured by Santa Cruz Biotechnology

Sc-1657 is a laboratory instrument designed for cell culture applications. It serves as an incubator for maintaining optimal environmental conditions for cell growth and proliferation.

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Lab products found in correlation

4 protocols using sc 1657

Western Blot Analysis of Insulin Signaling

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Typically, 10 µg of protein was resolved by SDS–PAGE, transferred to PVDF membranes and immunoblotted as previously described [5 (link)]. Antibodies detecting TRARG1 (sc-292062 or sc-377025) and 14-3-3 (sc-1657) were from Santa Cruz Biotechnology. Antibodies against pHSL S660 (#4126), pp90RSK (#9344), p4EBP1 S65 (#9456), pTBC1D4 T642 (#4288S), pAKT S473 (#4051), pAKT T308 (#9275L), AKT (#4685), HA (#C29F4), GSK3α (#9338S), GSK3β (#9832S), pGSK3 Ser 9/21 (#8566S), pGS S641 (#3891) and Caveolin1 (#3267) were purchased from Cell Signaling Technology. Anti-α-tubulin (#T9026) was from Sigma–Aldrich. Antibody against TBC1D4 was produced as previously described [4 (link)].

Duan X., Norris D.M., Humphrey S.J., Yang P., Cooke K.C., Bultitude W.P., Parker B.L., Conway O.J., Burchfield J.G., Krycer J.R., Brodsky F.M., James D.E, & Fazakerley D.J. (2022). Trafficking regulator of GLUT4-1 (TRARG1) is a GSK3 substrate. Biochemical Journal, 479(11), 1237-1256.

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Quantifying 14-3-3 Isoform Phosphorylation

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Primary antibodies included rabbit polyclonal antibody against pan 14‐3‐3 isoforms (Abcam #ab6081, Cambridge, MA), mouse monoclonal antibody against pan 14‐3‐3 isoforms (Santa Cruz #sc‐1657, Dallas, TX), mouse monoclonal antibody against 14‐3‐3θ (Abcam #ab10439), rabbit polyclonal antibody against phospho‐S58 (pS58) 14‐3‐3 (Abcam #ab51109), rabbit polyclonal antibody against phospho‐S232 (pS232) 14‐3‐3θ (Abcam #ab63369), sheep polyclonal antibody against phospho‐S185 (pS185) 14‐3‐3 (ENZO Life Science #SA‐479, Farmingdale, NY), and rabbit monoclonal antibody against phospho‐S129 αsyn (Abcam #ab168381). Horseradish peroxidase‐conjugated secondary antibodies were goat anti‐rabbit and goat anti‐sheep (Jackson Immunoresearch, West Grove, PA). Fluorescent‐conjugated secondary antibodies used were IRDye 800CW donkey anti‐rabbit and IRDye 680RD donkey anti‐mouse (LI‐COR Biotechnology, Lincoln, NE).

McFerrin M.B., Chi X., Cutter G, & Yacoubian T.A. (2017). Dysregulation of 14‐3‐3 proteins in neurodegenerative diseases with Lewy body or Alzheimer pathology. Annals of Clinical and Translational Neurology, 4(7), 466-477.

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Antibody Validation Protocols for Cell Biology

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Antibodies to GluR2 (75–002, NeuroMab), Bassoon (ADI-VAM-PS003-F, Enzo Life Sciences), DsRed (632496, Clonetech), 14-3-3 (sc-1657, Santa Cruz Biotechnology), Tubulin (T5326, Sigma), NeuroD1 (ab60704, Abcam), HERC2 (612366, BD Bioscience), Flag (A2220, Sigma M2), ERK1/2 (9102, Cell Signaling), Hoechst (B2883, Sigma), Cre (69050, Millipore), UBC13 (37-1100, Invitrogen), HA (MMS-101P, Covance), Parp (Cell Signaling), GFP (A6455, Invitrogen/ab3970, Abcam), goat serum (G9023, Sigma), rabbit IgG (12-370, Millipore), and mouse IgG (sc-2025, Santa Cruz) were purchased. The NEURL4 antibody was kindly provided by Brian Dynlacht (NYU Cancer Institute).
A rabbit RNF8 antibody was raised against bacterially produced glutathione-S-transferase fusion protein containing 100–250 aa of RNF8 and purified by affinity chromatography.
Uncropped western blot images are shown in Supplementary Fig. 11.

Valnegri P., Huang J., Yamada T., Yang Y., Mejia L.A., Cho H.Y., Oldenborg A, & Bonni A. (2017). RNF8/UBC13 ubiquitin signaling suppresses synapse formation in the mammalian brain. Nature Communications, 8, 1271.

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14-3-3 Protein Immunoblot Analysis

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Nine μl Urea/SDS samples (insoluble proteins) were loaded on a 4–12% gradient gel. The membrane was probed with anti-14-3-3 (1:5000, SC-1657, Santa Cruz Biotechnology). The quantification was done with ImageJ.

Groh N., Bühler A., Huang C., Li K.W., van Nierop P., Smit A.B., Fändrich M., Baumann F, & David D.C. (2017). Age-Dependent Protein Aggregation Initiates Amyloid-β Aggregation. Frontiers in Aging Neuroscience, 9, 138.

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