Akt pan 11e7

Manufactured by Cell Signaling Technology

Sourced in United States, United Kingdom

Akt (pan) (11E7) is a mouse monoclonal antibody that recognizes the Akt protein, also known as Protein Kinase B (PKB). Akt is a serine/threonine-specific protein kinase that plays a key role in multiple cellular processes, including cell proliferation, growth, and survival. The Akt (pan) (11E7) antibody is designed to detect the Akt protein across all three isoforms (Akt1, Akt2, and Akt3).

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Pan-AKT (169 citations) Akt (11E7) (6 citations) Akt (pan) (11E7) rabbit mAb (4 citations) Rabbit anti-pan-AKT (11E7; (3 citations) Anti-pan-Akt (11E7) (3 citations)

4 protocols using akt pan 11e7

Immunoblotting Analysis of Signaling Pathways

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The following antibodies were purchased from Cell Signaling Technology: HIF-1α (D1S7W) (Rabbit mAb#36,169); c-Myc (D84C12) (Rabbit mAb #5605); Phospho-Akt (Thr308) (244F9) (Rabbit mAb #4056); Phospho-Akt (Ser473) (587F11) (Mouse mAb #4051); Akt (pan) (11E7) (Rabbit mAb #4685); p70 S6 Kinase (49D7) (Rabbit mAb #2708); Phospho-p70 S6 Kinase (Thr421/Ser424) (Antibody #9204); Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) (Rabbit mAb (HRP Conjugate) #8544); p44/42 MAPK (Erk1/2) (137F5) (Rabbit mAb #4695); FAK antibody ( #3285); Phospho-FAK (Tyr576/577) antibody (#3281); CDK4 (D9G3E) (Rabbit mAb #12790); Rb (4H1) (Mouse mAb #9309); Phospho-Rb (Ser807/811) (D20B12) (Rabbit mAb #8516); Cyclin D1 antibody (#2922), PRAS40 antibody (#2610); Phospho-PRAS40 (Thr246) (D4D2) (Rabbit mAb #13175). The followings reagents were purchased from ABCAM: Anti-LOX antibody (ab174316); Anti-TGF beta 1 antibody (ab27969); Anti-Ki67 antibody (ab15580). rLOX was purchased from OriGene Technologies. BAPN was purchased from Sigma-Aldrich.

Li Q., Zhu C.C., Ni B., Zhang Z.Z., Jiang S.H., Hu L.P., Wang X., Zhang X.X., Huang P.Q., Yang Q., Li J., Gu J.R., Xu J., Luo K.Q., Zhao G, & Zhang Z.G. (2019). Lysyl oxidase promotes liver metastasis of gastric cancer via facilitating the reciprocal interactions between tumor cells and cancer associated fibroblasts. EBioMedicine, 49, 157-171.

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Western Blot Analysis of Cellular Proteins

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Total and cytosolic proteins were extracted and quantified as previously described [22 (link)], separated on SDS-PAGE gel, and transferred to nitrocellulose membranes. Proteins were detected using the following specific monoclonal antibodies (mAbs): FoxO3a (75D8, #2497), p-FoxO3a (Ser253; #13129), integrin α5 (#4705), pAkt (Ser 473, #4060), AKT (pan) (11E7, #4685), GAPDH (14C10 #2118) (all from Cell Signaling Technology, Danvers, MA, USA), β−Actin (AC-15) (Sigma-Aldrich, Merck, Milan, Italy), Lamin B1 (Invitrogen #702972) and IRDye secondary Abs (LI-COR Biosciences GmbH, Bad Homburg, Germany). Odyssey FC Imaging System was used for image acquisitions. Protein bands were quantified by means of Image Studio™ Lite v5.2 (LI-COR Biosciences GmbH, Bad Homburg, Germany). All “Original blots and densitometries” have been included in a separate file as Supplementary Information.

Ricci E., Fava M., Rizza P., Pellegrino M., Bonofiglio D., Casaburi I., Lanzino M., Giordano C., Bruno R., Sirianni R., Barone I., Sisci D, & Morelli C. (2022). FoxO3a Inhibits Tamoxifen-Resistant Breast Cancer Progression by Inducing Integrin α5 Expression. Cancers, 14(1), 214.

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Western Blot Analysis of XO, Akt, and Phospho-Akt

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After experiments, CMECs were homogenized in RIPA lysis buffer (Beyotime, China) containing 1 × phosphatase inhibitor cocktail (Cell Signaling Technology, MA, USA). Protein (40μg) was separated by SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with primary antibodies at 4°C overnight. Antibodies for XO (Abcam, Cambridge, UK), Akt (pan) (11E7) (Cell Signaling Technology, MA, USA), phosphor-Akt (Ser473) (D9E) (Cell Signaling Technology, MA, USA) at the concentration of 1:1000 were used. Horseradish peroxidase-conjugated secondary antibodies at the concentration of 1:5000 were incubated for 1 h at 37°C. The immunoreactive proteins on the membrane were detected by using enhanced chemiluminescence (Beyotime, China). The signal density of respective bands were qualified and normalized to GAPDH expression.

Zhang Y.Q., Tian F., Chen J.S., Chen Y.D., Zhou Y., Li B., Ma Q, & Zhang Y. (2016). Delayed reendothelialization with rapamycin is rescued by the addition of nicorandil in balloon-injured rat carotid arteries. Oncotarget, 7(46), 75926-75939.

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MSLN CAR-T Cells Transwell Invasion Assay

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FITC- or PE-conjugated mAbs (1:100 dilution) were used for staining and analyzed using a FACS Fortessa flow cytometer.
For Western blot analysis, β-Actin (8H10D10) (catalog# 3700T) mouse mAb and Cleaved Caspase-3 Ab (catalog# 4926), Cleaved Caspase-1 (catalog# 89332), RIP (E8S7U) XP^® (RIPK1, catalog# 73271), CD71 (D7G9X) XP® (TFRC, catalog# 13113), Phospho-Akt (Ser473) (D9E) XP® (catalog# 4060), and Akt (pan) (11E7) (catalog# 4685) rabbit mAbs from Cell Signaling Technology were used at a 1:1000 dilution. Anti-PI 3-kinase p100 (F-11) (catalog# sc-365404) and anti-alpha 1 sodium potassium ATPase/ATP1A1 (F-2) (catalog# sc-514614) mouse mAbs from Santa Cruz Biotechnology were used at a 1:500 dilution. In some experiments, SKOV3 tumor cells were seeded into upper (2×10⁵) and lower (2×10⁴) Transwell chambers with 400 μl of the completed medium. MSLN CAR-T (2×10⁴) or MSLN CAR-T^10%MYXV (2×10⁴) cells were added to the upper Transwell chamber. The pore size of Transwell inserts is 0.4 μM. After 24 hrs of coculture, the SKOV3 cells in the upper (after T cell removal) and lower chambers were lysed and used for Western blot analyses.

Zheng N., Fang J., Xue G., Wang Z., Li X., Zhou M., Jin G., Rahman M.M., McFadden G, & Lu Y. (2022). Induction of tumor cell autosis by myxoma virus-infected CAR-T and TCR-T cells to overcome primary and acquired resistance. Cancer cell, 40(9), 973-985.e7.

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