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Anti phospho raptor

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Anti-phospho-raptor is a primary antibody that detects the phosphorylated form of the raptor (regulatory associated protein of mTOR) protein. Raptor is a critical component of the mTOR complex 1 (mTORC1), which plays a key role in regulating cell growth, proliferation, and metabolism.

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Lab products found in correlation

Anti raptor, by Cell Signaling Technology (3 mentions) Anti phospho ampkα, by Cell Signaling Technology (2 mentions) Anti s6k, by Cell Signaling Technology (2 mentions) Anti ampkα, by Cell Signaling Technology (2 mentions) Anti crbn, by Merck Group (1 mentions) Anti lamin b, by Santa Cruz Biotechnology (1 mentions) Anti orai1, by Santa Cruz Biotechnology (1 mentions) Anti stim1, by Abcam (1 mentions) Anti phospho s6k, by Cell Signaling Technology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti cd16 32, by BioLegend (1 mentions) Anti orai1, by Alomone (1 mentions) Anti orai2, by Abcam (1 mentions) Anti cul4a, by Abcam (1 mentions) Anti ddb1, by Fortis Life Sciences (1 mentions) Anti erk2, by Santa Cruz Biotechnology (1 mentions) Anti ha, by Santa Cruz Biotechnology (1 mentions) Anti gst, by Santa Cruz Biotechnology (1 mentions) Anti flag, by Merck Group (1 mentions) Anti cd11b, by BioLegend (1 mentions)

4 protocols using anti phospho raptor

1

Molecular Mechanisms of CRAC Channel Regulation

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All plasmids used in this study were sequenced to confirm their identity. Crbn, Ampk, Slo, or HA-Ubiquitin constructs used in this study were described previously26 (link). Orai1 (MMM1013-202764440), Orai2 (MMM1013-202859855), and Orai3 (MMM1013-202842392) cDNA were purchased from Open Biosystems to construct expression vectors by a PCR-based strategy. Glutamine synthetase (GS) cDNA were synthesized from mRNA of the testis of mice and GS expression vectors were generated. The complete list of all primer sequences used in the study is provided as a Supplementary table. Anti-Flag (Sigma, F1804), anti-HA (Santa Cruz, sc-7392), anti-GST (Santa Cruz, sc-138), anti-Crbn (Sigma, HPA045910), anti-Orai1 (Santa Cruz, sc-68895), anti-Orai1 (Alomone labs, ACC-062), anti-Orai2 (Abcam, ab180146), anti-Ampkα (Cell signaling, #2532), anti-phospho-AMPKα (cell signaling, #2535), anti-Stim1 (Abcam, ab108994), anti-Cul4A (Abcam, ab92554), anti-DDB1 (Bethyl Laboratories, A300-462A), anti-phospho-raptor (Cell signaling, #2083), anti-raptor (Cell signaling, #2280), anti-phospho-S6K (Cell signaling, #9206), anti-S6K (Cell signaling, #9202), anti-Lamin B (Santa Cruz, sc-374015), anti-Erk2 (Santa Cruz, sc-154), anti-CD16/32 (BioLegend, #101302), anti-CD11b (BioLegend, #1010207), and anti-β-Actin (Santa Cruz, sc-1616) were purchased.
Moon H., Min C., Kim G., Kim D., Kim K., Lee S.A., Moon B., Yang S., Lee J., Yang S.J., Cho S.K., Lee G., Lee C.S., Park C.S, & Park D. (2020). Crbn modulates calcium influx by regulating Orai1 during efferocytosis. Nature Communications, 11, 5489.
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2

Phospho-AMPK Thr 172 Quantification

Cited 1 time
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Slides were prepared from isolated pancreata and visualized as previously detailed (11 (link)). ImageJ software was used to calculate the mean intensity on phospho-AMPK Thr 172 in the β-cell area and in the acinar tissue surrounding the islets. Western blotting was performed as previously described (11 (link)) with 100–150 human or mouse islets. Anti–acetyl-CoA-carboxylase (ACC), anti-phospho-ACC, anti-AMPK, anti–phospho-AMPK Thr 172, anti-Raptor, and anti–phospho-Raptor were from Cell Signaling Technology (Danvers, MA, USA). Anti-glucagon was from Sigma-Aldrich (St. Louis, MO, USA).
Martinez-Sanchez A., Nguyen-Tu M.S., Cebola I., Yavari A., Marchetti P., Piemonti L., de Koning E., Shapiro A.M., Johnson P., Sakamoto K., Smith D.M., Leclerc I., Ashrafian H., Ferrer J, & Rutter G.A. (2018). MiR-184 expression is regulated by AMPK in pancreatic islets. The FASEB Journal, 32(5), 2587-2600.
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3

Cell Lysis and Western Blot Analysis

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Cells were collected and lysed with RIPA buffer (1% NP40, 0.5% Sodium Deoxycholate, 0.1% SDS, Tris pH8.0) with protease inhibitors (Complete, Roche) or SDS Buffer (20% SDS, Tris pH6.8) with protease and phosphatase inhibitors (Roche). Western blots were probed with anti-p53 (1:500 CM5, Vector Labs), anti-LC3 (1:2000, NB100-2220, Novus), anti-S6K (1:1000, 9206, Cell Signaling Technology), anti-P-S6k (1:1000, 2708, Cell Signaling Technology), anti-4EBP1 (1:1000, 9452, Cell Signaling Technology), anti-P-4EBP1 (1:1000, 9459, Cell Signaling Technology), anti-SIVA (1:500, J.V.N, see below), anti-phospho-S6 (1:1000, 4858, Cell Signaling Technology), anti-S6 (1:1000, 2217, Cell Signaling Technology), anti-phospho-RAPTOR (1:1000, 2083, Cell Signaling Technology), anti-RAPTOR (1:1000, 2280, Cell Signaling Technology), anti-phospho-TSC2 (1:1000, 3617, Cell Signaling Technology), anti-TSC2 (1:1000, 3612, Cell Signaling Technology), anti-PARP (1:1000, 9532, Cell Signaling Technology) or anti-ACTIN (1:30,000 Sigma A2228). SIVA antibodies were generated by injecting rabbits with a recombinant mouse MBP-SIVA fusion protein and purified using recombinant GST-SIVA.
Van Nostrand J.L., Brisac A., Mello S.S., Jacobs S.B., Luong R, & Attardi L.D. (2015). The p53 Target Gene Siva Enables Non-Small Cell Lung Cancer Development. Cancer discovery, 5(6), 622-635.
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4

Western Blotting Analysis of Signaling Pathways

Cited 1 time
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Cell lysates were prepared and western blotting was performed as described previously (42 (link)). Proteins were separated using 6%, 10%, or 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were detected using the following primary antibodies: anti-phospho-AMPKα (Cell Signaling Technology, clone 40H9), anti-AMPKα (Cell Signaling Technology), anti-phospho-mTOR (Cell Signaling Technology, clone D9C2), anti-mTOR (Cell Signaling Technology, clone 7C10), anti-β-actin (Cell Signaling Technology, clone 13E5), anti-DDIT4 (Proteintech), anti-phospho-Rictor (Cell Signaling Technology, clone D30A3), anti-Rictor (Cell Signaling Technology, clone 53A2), anti-phospho-FoxO3a (Cell Signaling Technology, clone D18H8), anti-FoxO3a (Cell Signaling Technology, clone D19A7), anti-phospho-AKT (Cell Signaling Technology, clone Ser473), anti-AKT (Cell Signaling Technology, clone Ser473), anti-phospho-p70 S6 Kinase (Thr389) (Cell Signaling Technology, 9205S), anti p70 S6 Kinase (Cell Signaling Technology.9202S), anti-phospho-Raptor(Ser792) (Cell Signaling Technology, 2083S), and Raptor (24C12) (Cell Signaling Technology.2280S) antibodies. After the incubating with the corresponding secondary antibody, the ECL substrate (Thermo Fisher) signals were detected using an Amersham Imager 600 (Cytiva Life Sciences, GE HealthCare).
Kim H.E., Lee J.Y., Yoo D.H., Park H.H., Choi E.J., Nam K.H., Park J, & Choi J.K. (2024). Imidazole propionate ameliorates atopic dermatitis-like skin lesions by inhibiting mitochondrial ROS and mTORC2. Frontiers in Immunology, 15, 1324026.
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