Redd1 10638 1 ap

Manufactured by Proteintech

Sourced in United States

REDD1 (10638-1-AP) is a primary antibody product manufactured by Proteintech. It is designed to detect the REDD1 protein, also known as the regulated in development and DNA damage responses 1 protein. The REDD1 protein is involved in cellular processes and is commonly used in research applications.

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4 protocols using redd1 10638 1 ap

Coomassie and Western Blot Analysis

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A Coomassie protein assay was performed using Coomassie Plus Reagent (Thermo Scientific; Rockford, IL, USA). Western blotting was performed using a Bio-Rad mini-PROTEAN Tetra Cell system. Polyvinylidine difluoride (PVDF) membrane was purchased from Bio-Rad Laboratories (Hercules, CA, USA). Primary antibody for SREBP-1c (NB600-582) from NOVUS biologicals (Littleton, CO, USA), antibodies for phospho-p70 S6 Kinase Thr389 (9234), phospho-S6 Ribosomal Protein Ser240/244 (5364), phospho-MEK1/2 Ser217/221 (9154), phospho-ERK1/2 Thr202/Tyr204 (4370), cleaved caspase 3 (9664), LC3A/B (4108), and GAPDH (2118), were purchased from Cell Signaling Technology (Beverly, MA, USA), and REDD1 (10638-1-AP) was purchased from Proteintech. Enhanced chemiluminescence (ECL) reagent was purchased from Bio-Rad Laboratories (Clarity western ECL). Chemiluminescence imaging was performed on a Bio-Rad ChemiDoc MP Imager.

Dungan C.M., Li J, & Williamson D.L. (2016). Caloric Restriction Normalizes Obesity-Induced Alterations on Regulators of Skeletal Muscle Growth Signaling. Lipids, 51(8), 905-912.

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Signaling Pathway Regulation in Cells

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Fluocinolone acetonide (FA), croton oil (CO), Wortmannin (WM), AZD8055 (AZD), NVP-BEZ235 (NVP), MK-2206 (MK), and 5-bromo-2′-deoxyuridine (BrdU), were from Sigma Aldrich (St. Louis, MO), LY294002 (LY) was from LC Laboratories (Woburn, MA). TNF-α and cytoplasmic and nuclear fractionation kit were from Thermo Fisher Scientific (Waltham, MA).
We used antibodies to GR (sc-8992, RRID:AB_2155784), RelA/p65 (sc-8008, RRID:AB_628017), phospho-RelA/p65 (Ser536) (sc-136548, RRID:AB_10610391), IκB (sc-1643, RRID:AB_627772), phospho-IκB (Ser32) (sc-8404, RRID:AB_627773), FKBP51 (sc-271547, RRID:AB_10649040) (Santa Cruz Biotechnology, Dallas, TX); GAPDH (G9545, RRID:AB_796208, Sigma Aldrich); phospho-GR (Ser211) (4161, RRID:AB_2155797), phospho-rpS6 (Ser235/236) (4856S, RRID:AB_2181037), phospho-4E-BP1 (Thr37/46) (9459L, RRID:AB_2262165), phospho-P70S6K (Thr389) (9234, RRID:AB_2269803), phospho-AKT (Thr308 and Ser473) (9266S, RRID:AB_659801 and 9271, RRID:AB_329825), lamin B (12586, RRID:AB_2650517) and tubulin (2148, RRID:AB_2288042) (Cell Signaling, San Jose, CA), REDD1 (10638-1-AP, RRID,AB_2245711, Proteintech Group, Rosemont, IL).

Agarwal S., Mirzoeva S., Readhead B., Dudley J.T, & Budunova I. (2019). PI3K inhibitors protect against glucocorticoid-induced skin atrophy. EBioMedicine, 41, 526-537.

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Western Blot Analysis of Cellular Proteins

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Cell lysate was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. The protein was transferred to a nylon filter and was hybridized with antibody against AMPK (catalog number: #2532), phosphorylated AMPKα (Thr172) (#2535), 4E–BP1 (#9452), phosphorylated 4E–BP1 (#9459), p70 S6 kinase (p70S6K) (#9202), phosphorylated p70S6K (Thr389) (#9205), Bcl-2 (#2872), Bax (#2772), phosphorylated p53 (Ser15) (#9284), caspase-3 (#9668), cleaved caspase-3 (#9661), LC3A/B (#4108), Atg-5 (#2630), Beclin-1 (#3495) (Cell Signaling, Danvers, MA, USA), REDD1 (10638–1-AP) (Proteintech, Chicago, IL, USA), p53 (Ab-6, Clone DO-1) (Thermo Fisher Scientific, Fremont, CA, USA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab9484) (Abcam, Cambrige, UK) as a loading control followed by appropriate second antibody. Dimethyl sulfoxide (DMSO), a solvent for nutlin-3a, was also used as a control. The membranes were developed with the ECL system (GE Healthcare, Buckinghamshire, UK).

Shimazu K., Tada Y., Morinaga T., Shingyoji M., Sekine I., Shimada H., Hiroshima K., Namiki T., Tatsumi K, & Tagawa M. (2017). Metformin produces growth inhibitory effects in combination with nutlin-3a on malignant mesothelioma through a cross-talk between mTOR and p53 pathways. BMC Cancer, 17, 309.

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Analyzing mTOR Pathway Regulation

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The raptor antibody was from EMD Millipore (09-217); S6K1 pT389 (9234), S6K1 (2708), rpS6 pS235/S236 (2211), rpS6 (2217), Sestrin-2 (8487), ATF-4 (11815), raptor pS792 (2083), acetyl-CoA carboxylase (ACC) (3662), ACC pS79 (3661), and GCN2 (3302) antibodies were from Cell Signaling Technology (CST); ND1 (19703-1-AP), Sestrin2 (10795-1-AP), and Redd1 (10638-1-AP) antibodies were from Proteintech; HRI (MBS2538144) antibody was from MyBioSource; and the HSD17B10 (TA500724) antibody was from Life Technologies.

Condon K.J., Orozco J.M., Adelmann C.H., Spinelli J.B., Helm P.W.v.d., Roberts J.M., Kunchok T., & Sabatini D.M. (2020). Genome-wide CRISPR screens reveal multitiered mechanisms through which mTORC1 senses mitochondrial dysfunction.

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