Anti p akt1

Manufactured by Abcam

Sourced in United Kingdom, United States

Anti-p-Akt1 is a primary antibody that recognizes the phosphorylated form of Akt1 protein. Akt1 is a serine/threonine protein kinase that plays a key role in the regulation of cell growth, proliferation, survival, and metabolism. This antibody can be used to detect and quantify the levels of phosphorylated Akt1 in various biological samples.

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4 protocols using anti p akt1

Antibody-based Expression Analysis in Cancer Cells

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The following antibodies were used for immunofluorescence staining: anti-GAPDH (1:5,000; cat. no. Ab8245; Abcam); anti-UCP2 (1:100; cat. no. 11081-1-AP; ProteinTech Group, Inc.) and anti-FLNa (1:100; cat. no. ab76289; Abcam). The antibodies were used to detect UCP2 and FLNa expression in different CC cell lines.
The following antibodies were used for immunohistochemical staining: anti-UCP2 (1:200; cat. no. 11081-1-AP; ProteinTech Group, Inc.), anti-FLNa (1:200; cat. no. ab76289; Abcam), anti-P16 (1:200; cat. no. 10883-1-AP; ProteinTech Group, Inc.), and anti-Ki67 (1:1,000; cat. no. ab92742; Abcam).
The following primary antibodies were used for western blotting of relevant proteins: anti-UCP2 (1:1,000; cat. no. 11081-1-AP; ProteinTech Group, Inc.), anti-FLNa (1:1,000; cat. no. ab76289; Abcam), anti-GAPDH (1:5,000; cat. no. ab8245; Abcam), anti-ERK1/2 (1:1,000; cat. no. 4695; Cell Signaling Technology, Inc.), anti-P-ERK1/2 (1:1,000; cat. no. 4370; Cell Signaling Technology, Inc.), anti-AKT1 (1:1,000; cat. no. ab227100; Abcam), anti-P-AKT1 (1:10,000; cat. no. ab81283; Abcam), and anti-BCL-2 (1:1,000 cat. no. ab32124; Abcam).

Wang A., Liu L., Yuan M., Han S., You X., Zhang H., Lei F, & Zhang Y. (2020). Role and mechanism of FLNa and UCP2 in the development of cervical cancer. Oncology Reports, 44(6), 2656-2668.

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Protein Expression Analysis in HCT-116 Cells

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HCT-116 cells were collected and lysed in radio immunoprecipitation assay (RIPA) lysis buffer (Sigma-Aldrich) containing protease inhibitors and phosphatase. Protein concentration of the lysates was quantified using the bicinchoninic acid Protein Assay Kit (Jiancheng Bioengineering Institute, Nanjing, Jiangsu, China). The proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) at 110 V for 2 h and then transferred onto a polyvinylidene fluoride membrane (Millipore, Boston, MA, USA) at 60 V for 2 h. After blocking with 5% nonfat milk at room temperature for 1 h, the polyvinylidene fluoride membranes were incubated with anti-Bcl-2 (1:1,000; Abcam, Cambridge, UK), anti-Bax (1:1,000; Abcam), anti-p-Akt1 (1:1,000; Abcam), anti-P-ERK (1:1,000; Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1,000; Abcam) primary antibodies overnight at 4°C. The polyvinylidene fluoride membranes were washed with tris buffered saline tween (TBST) and incubated with a horseradish peroxidase–conjugated secondary antibody at room temperature for 45 min. Visualization was performed with a chemiluminescent detection kit (Millipore). The densitometry of the bands was analyzed using Gel-Pro 6.3 software (Media Cybernetics, Inc., Bethesda, MD,USA).

Bai J.H., Xu J., Zhao J, & Zhang R. (2020). Ganoderma lucidum Polysaccharide Enzymatic Hydrolysate Suppresses the Growth of Human Colon Cancer Cells via Inducing Apoptosis. Cell Transplantation, 29, 0963689720931435.

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Proteomic Analysis of BMSC Proteins

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Proteins were extracted from more than 10⁶ harvested BMSCs according to the kit protocol (KeyGen Biotech, China). [46 (link)] BMSCs were digested in a lysis buffer cocktail and centrifuged at 12000 g for 15 minutes at 4 °C to get the supernatant as the total protein. 10% sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) was used for electrophoresis of protein (20 μg) with loading buffer and then blotted to a polyvinylidene fluoride (PVDF) membrane (Millipore Co., USA). Membranes were blocked with 5% skimmed milk in Tris-buffered saline tween 20, TBST, and then incubated overnight at 4°C with primary antibodies; anti-IHH, anti-P53, anti-P16, anti-PI3K, anti-p-PI3K, anti-Akt 1, anti-p-Akt 1, anti-NF-κB, anti-p-NF-κB-p65, anti-STAT3, anti-p-STAT3, anti-4EBP1, anti-p- 4EBP1, anti-p70S6K, anti-p-p70S6K and anti-β-actin (1:1000), all from Abcam, USA. The membranes were incubated for 1hour at RT with secondary antibody conjugated with horseradish peroxidase. Finally, TBST-washed membranes were treated with enhanced chemiluminescent (ECL) for detection (Bio-Rad, USA). Image Lab detection system (Bio-Rad, USA) was used for protein bands imaging and analysis.

Al-Azab M., Wang B., Elkhider A., Walana W., Li W., Yuan B., Ye Y., Tang Y., Almoiliqy M., Adlat S., Wei J., Zhang Y, & Li X. (2020). Indian Hedgehog regulates senescence in bone marrow-derived mesenchymal stem cell through modulation of ROS/mTOR/4EBP1, p70S6K1/2 pathway. Aging (Albany NY), 12(7), 5693-5715.

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Immunohistochemical Analysis of Ectopic Endometria

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Rats’ ectopic endometria fixed in a 4% neutral paraformaldehyde solution, were washed, dehydrated, waxed, and embedded. Slides, soaked in a wash buffer over-night, were rinsed with running water and dried. After being immersed in 100 pg/ml polylysine for 30 min at 37°C, the slides were inserted into a clean glass shelf overnight at 37°C. A microtome was used to cut 4-μm-thick serial sections, and 5 of these sections were selected to sit for 1 h at 56°C and then over-night at 37°C. Before staining, the sections were placed in a 60°Cwarm case for 1 h. Sections were then dewaxed in xylene and rehydrated, followed by microwaving in 10 mM sodium citrate buffer (pH 6.0) for antigen retrieval. The sections were washed three times with PBS (0.01 mmol/L, pH 7.4) for 3 min. The immunohistochemistry S-P method was conducted according to the manufacturer’s instructions (Maixin-Bio Corporation, Fuzhou, China). The dilution ratio of anti-VEGF, anti-VEGF-2 and anti-p-Akt1(Abcam) was 1:200. The dilution ratio of anti-Akt1, anti-mTOR(Abcam), and anti-p-mTOR (Cell Signaling Technology) was 1:100.

Cao Y., Ye Q., Zhuang M., Xie S., Zhong R., Cui J., Zhou J., Zhu Y., Zhang T, & Cao L. (2017). Ginsenoside Rg3 inhibits angiogenesis in a rat model of endometriosis through the VEGFR-2-mediated PI3K/Akt/mTOR signaling pathway. PLoS ONE, 12(11), e0186520.

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