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Phospho akt1

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Phospho-Akt1 is a laboratory reagent used for the detection and quantification of phosphorylated Akt1 protein. Akt1 is a serine/threonine-specific protein kinase that plays a key role in cellular processes such as glucose metabolism, apoptosis, cell proliferation, transcription, and cell migration. The Phospho-Akt1 reagent allows researchers to measure the activation state of Akt1 in biological samples.

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Lab products found in correlation

Phospho akt2, by Cell Signaling Technology (3 mentions) Vdac 4866, by Cell Signaling Technology (3 mentions) Gsk3α β, by Cell Signaling Technology (2 mentions) β actin a5441, by Merck Group (2 mentions) Phospho gsk3α β, by Cell Signaling Technology (2 mentions) Pgc 1α, by Cell Signaling Technology (2 mentions) Phospho pgc 1α s571 af6650, by R&D Systems (2 mentions) P38 sc 7972 and tom20 sc 11415, by Santa Cruz Biotechnology (2 mentions) Spc ab3786, by Merck Group (2 mentions) Akt1 2, by Cell Signaling Technology (2 mentions) Phospho drp1, by Cell Signaling Technology (2 mentions) Drp1 611113, by BD (2 mentions) Parkin, by Cell Signaling Technology (2 mentions) Phospho p38 mapk, by Cell Signaling Technology (2 mentions) Phospho stat3, by Cell Signaling Technology (2 mentions) Phospho erk1 2, by Cell Signaling Technology (2 mentions) β actin, by Merck Group (1 mentions) Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor, by Merck Group (1 mentions) Gapdh, by Bio-Rad (1 mentions)

Close spelling variants of this product

Phospho-AKT1 (Ser473), (7 citations) PathScan Phospho-Akt1 (Ser473) Sandwich ELISA Kit (6 citations) Anti-phospho-AKT1 (Ser473) (5 citations) Phospho-AKT1 (S473) (4 citations) Anti-phospho-Akt1 (4 citations) Pathscan phospho-Akt1 sandwich ELISA kit (2 citations)

13 protocols using phospho akt1

1

Multimodal Mitochondrial Function Analysis

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Primary antibodies used: Akt1 (2938), Akt2 (5239), Akt1/2 (9272), GSK-3α/β (5676), LC3 (4108S), Lamin A/C (2032), NFR1 (12381), PGC-1α (2178), phospho-p38 MAPK (9215), phospho-Akt1 (9018), phospho-Akt2 (8599), phospho-DRP1 (Ser616) (4494), phospho-GSK-3α/β (9331), Parkin (2132S), Tfam (7495S), VDAC (4866) (Cell Signaling); β-actin (A5441) (Sigma); PINK1 (ab23707), Mfn-2 (ab124773) (Abcam); Drp1 (611113) (BD Biosciences); phospho-PGC-1α (S571) (AF6650) (R&D Systems); p38 (sc-7972) and Tom20 (sc-11415) (Santa Cruz); SPC (AB3786) (Millipore).
Larson‐Casey J.L., Gu L., Davis D., Cai G., Ding Q., He C, & Carter A.B. (2021). Post‐translational regulation of PGC‐1α modulates fibrotic repair. The FASEB Journal, 35(6), e21675.
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2

Mitochondrial Dynamics and Signaling Pathways

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Primary antibodies used: Akt1 (2938), Akt2 (5239), Akt1/2 (9272), GSK‐3α/β (5676), LC3 (4108S), Lamin A/C (2032), NFR1 (12381), PGC‐1α (2178), phospho‐p38 MAPK (9215), phospho‐Akt1 (9018), phospho‐Akt2 (8599), phospho‐DRP1 (Ser616) (4494), phospho‐GSK‐3α/β (9331), Parkin (2132S), Tfam (7495S), VDAC (4866) (Cell Signaling); β‐actin (A5441) (Sigma); PINK1 (ab23707), Mfn‐2 (ab124773) (Abcam); Drp1 (611113) (BD Biosciences); phospho‐PGC‐1α (S571) (AF6650) (R&D Systems); p38 (sc‐7972) and Tom20 (sc‐11415) (Santa Cruz); SPC (AB3786) (Millipore).
Larson‐Casey J.L., Gu L., Davis D., Cai G., Ding Q., He C, & Carter A.B. (2021). Post‐translational regulation of PGC‐1α modulates fibrotic repair. The FASEB Journal, 35(6), e21675.
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3

Western Blot Analysis of Signaling Proteins

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Cells were lysed in 1× Laemmli buffer supplemented with phosphatase inhibitors (Sigma-Aldrich) and 1× complete protease inhibitor cocktail (Roche; Indianapolis, IN), size fractionated on 12% acrylamide gels and subsequently electro-transferred onto Hybond enhanced chemiluminescent membranes (GE Healthcare; Waukesha, WI). Membranes were blocked in 5% milk in tris-buffered saline containing 1% tween 20 and probed with antibodies against phospho-jun proto-oncogene (pJUN; pc-Jun) (9164; Cell Signaling Technology; Danvers, MA), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (NFKBIA; IκBα) (sc-371; Santa Cruz Biotechnology; Dallas, TX), phospho-NFKBIA (9246; Cell Signaling Technology), phospho-AKT1 (9271S; Cell Signaling Technology), AKT (9272S; Cell Signaling Technology) and GAPDH (4699–9555(ST); AbD Serotec; Raleigh, NC). After washing, membranes were incubated with horseradish peroxidase-conjugated anti-rabbit (111–035–003; Jackson ImmunoResearch Laboratories Inc; West Grove, PA) or anti-mouse immunoglobulin (115–035–003; Jackson ImmunoResearch Laboratories Inc). Detection was by enhanced chemiluminescence (Pierce ECL Western Blotting Substrate; Thermo Fisher Scientific Inc.; Rockford, IL) and visualization by autoradiography.
Rider C.F., Shah S., Miller-Larsson A., Giembycz M.A, & Newton R. (2015). Cytokine-Induced Loss of Glucocorticoid Function: Effect of Kinase Inhibitors, Long-Acting β2-Adrenoceptor Agonist and Glucocorticoid Receptor Ligands. PLoS ONE, 10(1), e0116773.
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4

Analyzing Vascular Signaling Proteins

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The venous limb of the AVF was harvested and treated with RIPA lysis buffer containing protease and phosphatase inhibitors (Roche). Equal amounts of protein were loaded and run in SDS-PAGE followed by Western blot analysis. Protein expression was probed with the following antibodies: Cav-1 (BD Biosciences, 610406), phospho-eNOS (Cell Signaling Technology, 9571), eNOS (Santa Cruz, sc-654), phospho-Akt1 (Cell Signaling Technology, 9018), Akt1 (Cell Signaling Technology, 2967), or GAPDH (Cell Signaling Technology, 2118). Membrane signals were detected using ECL detection reagent (GE Healthcare, Denville scientific).
Hashimoto T., Isaji T., Hu H., Yamamoto K., Bai H., Santana J.M., Kuo A., Kuwahara G., Foster T.R., Hanisch J.J., Yatsula B.A., Sessa W.C., Hoshina K, & Dardik A. (2019). Stimulation of Caveolin-1 Signaling Improves Arteriovenous Fistula Patency. Arteriosclerosis, thrombosis, and vascular biology, 39(4), 754-764.
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5

Osteoclastogenesis Regulation by ISO

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ISO was purchased from MedChem Express (Monmouth Junction, NJ, USA), dissolved in dimethyl sulfoxide (DMSO; Beyotime) at the stock concentration of 20 mM, and diluted with culture medium to form working concentrations prior to use. Recombinant human RANKL was obtained from PeproTech EC, Ltd. Minimum Essential Medium Alpha (α-MEM), Dulbecco’s modified Eagle’s medium (DMEM), and fetal bovine serum (FBS) were purchased from Gibco. The CCK8 assay kit and tartrate‐resistant acid phosphatase (TRAP) assay kit were obtained from Beyotime. The TRAP staining kit was provided by Sigma‐Aldrich. The primary antibodies of NFATc1 and c-Fos were acquired from ABclonal. The primary antibodies for β-Actin, P38, phospho-P38, extracellular signal-egulated kinase (ERK), phospho-ERK, c-Jun N-terminal kinase (JNK), phospho-JNK, Nuclear factor erythroid 2‐related factor 2 (Nrf2), heme oxygenase‐1 (HO-1), catalase (CAT), and γ‐glutamyl cysteine synthetase catalytic subunit (GCLC) were acquired from Beyotime. The primary antibodies for phosphoinositide 3-kinase (PI3K), phospho-PI3K, Protein Kinase B α (AKT1), and phospho-AKT1 were purchased from Cell Signaling Technology.
Zhang B., Qu Z., Hui H., He B., Wang D., Zhang Y., Zhao Y., Zhang J, & Yan L. (2024). Exploring the therapeutic potential of isoorientin in the treatment of osteoporosis: a study using network pharmacology and experimental validation. Molecular Medicine, 30, 27.
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6

Western Blot Analysis of AKT Signaling

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Protein lysates from human or mouse heart samples were prepared in RIPA buffer supplemented with a cocktail of protease and phosphatase inhibitors (Cell Signaling). Equal amounts of total proteins were loaded and separated in SDS-polyacrylamide gel. After electrophoresis, the proteins were electrotransferred onto nitrocellulose membranes (0.2 μm, GE Healthcare) and incubated for 1 h with 5% bovine serum albumin followed by overnight incubation at 4 °C with primary antibody. Finally, the membranes were incubated for 1 h with secondary antibodies conjugated to horseradish peroxidase. Immunoblots were visualized with the Western Blotting Chemiluminescence Luminol (Pierce, Thermo Fisher Scientific Inc.) using the ChemiDoc™ XRS System (Bio-Rad). The primary antibodies used in the study were: phospho-AKT1 (1:600; Cell Signaling, cat. 9018); total AKT1 (1:1000; Cell Signaling, cat. 2938); phospho AKT2 (1:600, Cell Signaling, cat. 8599); total AKT2 (1:1000; Cell Signaling, cat. 3063); and GAPDH (1: 5000; Sigma-Aldrich, cat. G9545). Full scans of the original uncropped western blots are shown in Supplementary Figs. 9 and 10.
Silva M.C., Davoli-Ferreira M., Medina T.S., Sesti-Costa R., Silva G.K., Lopes C.D., Cardozo L.E., Gava F.N., Lyroni K., Dias F.C., Frade A.F., Baron M., Nakaya H.I., Figueiredo F., Alves-Filho J.C., Cunha F.Q., Tsatsanis C., Chevillard C., Cunha-Neto E., Hirsch E., Silva J.S, & Cunha T.M. (2018). Canonical PI3Kγ signaling in myeloid cells restricts Trypanosoma cruzi infection and dampens chagasic myocarditis. Nature Communications, 9, 1513.
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7

Immunostaining and Image Analysis of Mouse Aortas

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We performed immunofluorescence staining on 5-μm sections, using antibodies to Ki67 (Abcam #ab16667), smooth muscle α-actin (SMA; Sigma-Aldrich, #A5228, St. Louis, MO, USA), CD45 (BioLegend #103101, San Diego, CA, USA), phospho-Stat3 (Cell Signaling Technology #9145, Danvers, MA, USA), phospho-Akt1 (Cell Signaling Technology #4060), phospho-Akt2 (Abcam #ab38513, Cambridge, UK), B220 (BD #553086, Becton Dickinson, Franklin Lakes, NJ, USA), CD3 (Abcam #ab11089), Iba-1 (Millipore #MABN92, Burlington, MA, USA), and Ly-6G (Abcam #ab25377). Nuclei were visualized using DAPI staining. Imaging cytometric analysis of mouse aortas was performed using ArrayScan XTI (Thermo Fisher Scientific, Waltham, MA, USA) and FlowJo 10 software (Becton Dickinson). Two aortic tissue sections were obtained from each mouse in four experimental groups, namely DMSO, BA + DMSO, rapamycin, and BA+rapamycin, where each experimental group included three mice.
Hayashi-Hori M., Aoki H., Matsukuma M., Majima R., Hashimoto Y., Ito S., Hirakata S., Nishida N., Furusho A., Ohno-Urabe S, & Fukumoto Y. (2020). Therapeutic Effect of Rapamycin on Aortic Dissection in Mice. International Journal of Molecular Sciences, 21(9), 3341.
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8

Protein Extraction and Western Blot Analysis

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Cells have been lysed using M-PER reagent (Thermo Scientific); protein concentration was measured by a BCA protein assay (Thermo Scientific). Cellular lysates have been denatured in reducing SDS-loading buffer, separated by SDS-PAGE (Bio-Rad) and transferred to nitrocellulose membrane by wet blotting. vGPCR protein levels were assessed using an anti-myc antibody (Biolegend, #626802). Additionally used antibodies: leptin receptor (US Biological, #L1671-03R), phosho-Jak2 (#3776), phospho-Stat3 (#9145), phospho-Akt1 (#4060), phospho-Erk1/2 (#4377), all Cell Signaling Technology, β-actin (Abcam, #ab6276) and secondary HRP-conjugated antibodies from Jackson Immunoresearch. Western blots have been developed using ECL reagent (GE Healthcare).
Krause C.J., Popp O., Thirunarayanan N., Dittmar G., Lipp M, & Müller G. (2016). MicroRNA-34a promotes genomic instability by a broad suppression of genome maintenance mechanisms downstream of the oncogene KSHV-vGPCR. Oncotarget, 7(9), 10414-10432.
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9

Apoptosis Signaling Pathway Profiling

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Primary antibodies used: Akt1 (2938), Bad (9239), Lamin A/C (2032), Bax (2772), phospho-Akt1 (9018), phospho-Bad (Ser136) (4366), VDAC (4866) (Cell Signaling); β-actin (A5441) (Sigma); p38 (sc-7972) (Santa Cruz); NOX4 (NB110-58849) (Novus), NOX4 (109225) (Abcam); Bak (A0404) (Abclonal).
Larson-Casey J.L., Gu L., Kang J., Dhyani A, & Carter A.B. (2021). NOX4 regulates macrophage apoptosis resistance to induce fibrotic progression. The Journal of Biological Chemistry, 297(1), 100810.
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10

Molecular Mechanisms of Melanoma Regulation

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Melanoma cell lines G-361, SK-Mel-28, SK-Mel-30, RPMI-7951, and normal human primary epidermal melanocytes were all purchased from American Type Culture Collection (ATCC) (Manasas, VA), and were authenticated according to the ATCC guidelines. I3C and MG-132 were purchased from Sigma Aldrich (St. Louis, MO). Two different PTEN siRNAs, two distinct NEDD4-1 siRNAs and the corresponding scrambled siRNAs, and the HiPerFect reagents were purchased from Qiagen (Valencia, CA). Antibodies to PTEN, NEDD4-1, ubiquitin, Bcl-2 and MDM-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA), the antibody to Hsp90 was purchased from BD Biosciences Franklin (Lakes, NJ) and antibodies to PARP, cleaved PARP, cleaved caspase 3, p53, phosphor-MDM2, Phospho-AKT-1, and AKT-1 were acquired from Cell Signaling Technology (Beverly MA). The secondary anti-mouse and anti-rabbit antibodies conjugated with HRP were obtained from Bio-Rad Laboratories (Hercules, CA). Secondary antibodies conjugated to fluorescent probes were purchased from Molecular Probes/Invitrogen (Eugene, OR). All primers used in RT-PCR reactions were synthesized by IDT technologies (San Diego, CA.)
Aronchik I., Kundu A., Quirit J.G, & Firestone G.L. (2014). The Anti-proliferative Response of Indole-3-carbinol in human melanoma cells is Triggered by an Interaction with NEDD4-1 and Disruption of Wild-type PTEN Degradation. Molecular cancer research : MCR, 12(11), 1621-1634.
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