Akt pan 11e7 rabbit mab

Manufactured by Cell Signaling Technology

Sourced in United States

Akt (pan) (11E7) rabbit mAb is a monoclonal antibody that recognizes the Akt protein. Akt is a serine/threonine protein kinase that plays a key role in the regulation of cellular processes such as cell proliferation, survival, and metabolism.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (1 mentions) Odyssey clx imager, by LI COR (1 mentions) Irdye 680rd donkey anti mouse igg, by LI COR (1 mentions) Phospho akt ser473 antibody, by Cell Signaling Technology (1 mentions) β actin antibody, by Merck Group (1 mentions) Phospho p70 s6 kinase thr421 ser424, by Cell Signaling Technology (1 mentions) Ube3a, by Abcepta (1 mentions) Anti psd 95 antibody, by Abcam (1 mentions) Nitrocellulose membrane, by Merck Group (1 mentions) Immobilon western chemiluminescent hrp substrate system, by Merck Group (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions) Ab50011, by Abcam (1 mentions) Ab17942, by Abcam (1 mentions) Goat anti rabbit igg h l hrp conjugate, by Bio-Rad (1 mentions) Iblot gel transfer system, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Pan-AKT (169 citations) Akt (11E7) (6 citations) Akt (pan) (11E7) (4 citations) Akt (pan) rabbit mAb (3 citations) 11E7 Akt rabbit mAb (2 citations) Rabbit mABs (2 citations)

4 protocols using akt pan 11e7 rabbit mab

Insulin Signaling Pathway Activation

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Insulin signaling pathway activation in mice was performed by administering 5 U/kg insulin intraperitoneally 15 min prior to the sacrifice of the mice. In control mice, saline was injected. Liver, epididymal fat, and skeletal muscle tissue were collected, snap frozen in liquid nitrogen, and stored at −80°C. Tissues were homogenized through the Ripa buffer (ThermoFisher, VH310061, Waltham, MA, USA). The gels for Western blot were made by a gel-making kit (Solarbio, PC0020). Proteins (30 μg) were applied to each lane. For total and phosphorylated Akt measurements, we used antibody phospho-Akt (Ser473) (D9E) XP rabbit mAb (Cell Signaling, #4060, Danvers, MA, USA) and Akt (pan) (11E7) rabbit mAb (Cell Signaling, #4685). The optical densities of bands were measured using the ImageJ software. Insulin signaling was assessed by calculation of the ratio of pAkt to actin or total Akt after insulin injection.

Rozjan A., Shan W, & Yao Q. (2021). Metabolic Consequences of Neuronal HIF1α-Deficiency in Mediobasal Hypothalamus in Mice. Frontiers in Endocrinology, 12, 668193.

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Protein Expression Profiling Assay

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NuPAGE Novex Midi Gels (4–12%) from Life Technologies were used for protein separation. The following antibodies were used: Anti-PSD95 antibody (Abcam, Cat. #ab18258), Synaptophysin (D35E4) XP^® Rabbit mAb (Cell Signaling, Cat. #5461), Complex II Subunit Monoclonal Antibody (clone 21A11AE7, Life Technology, #439454), Phospho-p70 S6 Kinase (Thr421/Ser424) (Cell Signaling, #9204), S6K(p70) (Cell Signaling, #2217), Phospho-Akt (Ser473) Antibody (Cell Signaling, Cat. #9271s), Akt (pan) (11E7) Rabbit mAb (Cell Signaling, Cat. #4685s), Ube3a (Abgent, #346954), β-Tubulin Antibody (TUB 2.1, Santa Cruz, Cat. #sc-58886) and β-Actin antibody (Sigma, Cat. #A1978). After incubating with corresponding infrared fluorescent secondary antibodies from Li_COR Biosciences (#926–32213 IRDye^® 800CW Donkey anti-Rabbit IgG, and #926–68072 IRDye^® 680RD Donkey anti-Mouse IgG), the signal of specific protein was detected and quantified on the Odyssey CLx imager (LI-COR).

Wang D, & Mitchell E.S. (2016). Cognition and Synaptic-Plasticity Related Changes in Aged Rats Supplemented with 8- and 10-Carbon Medium Chain Triglycerides. PLoS ONE, 11(8), e0160159.

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Western Blot Analysis of Protein Expression

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Protein samples (5–10 µg) were separated on 8–10% gels by SDS-PAGE and transferred onto a nitrocellulose membrane (Merck, Millipore). The membrane was then blocked and incubated with the desired primary antibodies diluted 1:1000: anti OPCML (goat pAb, #AF2777, R&D Systems), phospho-AKT (Thr308 D25E6 XP® rabbit mAb, #13038, Cell Signaling), AKT (pan 11E7 rabbit mAb, #4685 Cell Signaling), phospho-ERK1 (pT202/pY204) + phospho-ERK2 (pT185/pY187) (mouse mAb, ab50011, Abcam), ERK1/2 (rabbit pAb, ab17942, Abcam) and anti-GAPDH (Cell Signalling), over-night at 4 °C. The membrane was washed, then incubated with the appropriate secondary antibodies (DAKO) diluted 1:5000. After final washings, proteins were detected using Immobilon Western Chemiluminescent HRP Substrate system (Millipore) and GE Healthcare Amersham™ Hyperfilm™ ECL film with a Kodak SRX2000 (Rochester, NY, USA) developer machine.

Birtley J.R., Alomary M., Zanini E., Antony J., Maben Z., Weaver G.C., Von Arx C., Mura M., Marinho A.T., Lu H., Morecroft E.V., Karali E., Chayen N.E., Tate E.W., Jurewicz M., Stern L.J., Recchi C, & Gabra H. (2019). Inactivating mutations and X-ray crystal structure of the tumor suppressor OPCML reveal cancer-associated functions. Nature Communications, 10, 3134.

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Western Blot Analysis of Signaling Proteins

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The cells were lysed in a buffer of 20 mM HEPES, pH 7.4, 50 mM β-glycerophosphate, 1 mM Na₃VO₄, 0.5% (vol/vol) Triton X-100, 0.5% (vol/vol) CHAPS (3-[(3-cholamidopropyl)-dimethylammonio]-1- propane sulfonate hydrate), and 10% (vol/vol) glycerol with a protease inhibitor 'cocktail' tablet. The lysates were separated by SDS-PAGE and transferred to nitrocellulose membrane by iBLOT Gel Transfer System (Invitrogen). The membrane was incubated with 5% nonfat milk w/v in TBS buffer (25 mM Tris- HCl, pH 7.5; 150 mM NaCl; 0.1% Tween-20) for 1 h, and then reacted with the primary antibody in TBS buffer with 2.5 % nonfat milk or 5% BSA w/v for overnight by shaking at 4 degree. The appropriated second antibodies conjugated to HRP were used to detect the protein of interest via enhanced chemoluminescence (ECL). The following primary antibodies were used phosphor-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) mouse mAb #9106, 1:2000; phosospho-Akt (Ser473) (D9E) XP^® rabbit mAb #4060, 1:2000; p44/42 MAPK (Erk1/2) antibody #9102, 1:1000; Akt (pan) (11E7) rabbit mAb #4685, 1:1000 (all from Cell Signaling); Gα_i2 antibody (L5): sc-13534, mouse mAb, 1:500 (Santa Cruz). Secondary antibody for rabbit primaries was Goat anti-rabbit IgG (H + L)-HRP conjugate #170- 6515, 1:3000 (BioRad); and for the mouse primaries was horse anti-mouse IgG, HRP-linked antibody #7076, 1:3000, (Cell Signaling).

Hwang I.Y., Park C., Harrison K., Boularan C., Galés C, & Kehrl J.H. (2015). An Essential Role for RGS Protein/Gαi2 Interactions in B Lymphocyte Directed Cell Migration and Trafficking. Journal of immunology (Baltimore, Md. : 1950), 194(5), 2128-2139.

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