Ab170880

Manufactured by Abcam

Sourced in United States

Ab170880 is a laboratory equipment product. It serves as a core function related to the intended use of the product. No further details are available for this product in an unbiased and factual manner.

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Lab products found in correlation

3 protocols using ab170880

Western Blot Analysis of Phospho-Akt and Phospho-MDM2

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The cell monolayers were washed with PBS and lysed at the indicated times. The lysates were collected and incubated on ice for 10 min. The lysates were cleared by centrifugation at 10,000g for 5 min at 4 °C. The supernatants were analyzed for total protein content with a BCA protein-assay kit (Beyotime). The total protein (30 μg) was resolved by 10% SDS–PAGE and transferred onto nitrocellulose membranes (Millipore). The membranes were blocked with 5% skim milk for 1 h at 37 °C and then incubated overnight at 4 °C with specific antisera: rabbit anti-phospho-Akt antibody (Ser473) (ab81283, Abcam), rabbit anti-Akt antibody (ab32505, Abcam), rabbit anti-phospho-MDM2 antibody (Ser166) (ab170880, Abcam), rabbit anti-MDM2 antibody (ab38618, Abcam), rabbit PI3K p85 antibody (ab40755, Abcam), rabbit anti-glyceraldehyde 3-phosphate dehydrogenase (GADPH) antibody (ab22555, Abcam) and rabbit anti-p53 antibody (9282, CST). After three rinses in TBST buffer, the membranes were incubated at 37 °C for 1 h with IRDye 800DX-conjugated anti-rabbit IgG (1:8000; Rockland Immunochemicals) diluted in TBST as a secondary antibody. The membranes were washed three times in PBST, then visualized and analyzed with an Odyssey infrared imaging system (LI-COR Biosciences).

Zhang X., Wu H., Liu C., Tian J, & Qu L. (2015). PI3K/Akt/p53 pathway inhibits reovirus infection. Infection, Genetics and Evolution, 34, 415-422.

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Western Blot Profiling of Cellular Proteins

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Total proteins in cells were extracted using protein lysis solution (Tiangen Biotech, Beijing, China). The protein concentration was measured with a bicinchoninic acid kit (Tiangen Biotech). The protein extracts were resolved through sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, transferred onto polyvinylidene difluoride membranes (Bio-Rad, Berkeley, CA, USA), which were blocked with 5% non-fat milk solutions for 1 h at room temperature. The target protiens were then detected using primary antibodies against human TUSC3 (1:500; ab77600), signal transducer and activator of transcription 3 (STAT3; 1:1,000; ab109085), mouse double minute 2 homolog (MDM2; 1:500; ab170880), p53 (1:500; ab76242), MET (1:1,000; ab68141), CD133 (1:500; ab19898), zinc finger protein, X-linked (ZFX; 1:500; ab115998) and β-actin (1:500; ab8227) (all from Abcam, Cambridge, MA, USA) at at 4°C overnight. The blots were then washed with 0.1% Tween-PBS and incubated with goat anti-rabbit secondary antibodies (1:500; ab218695; Abcam) for antibody for 1 h at room temperature. The blots were then detected by enhanced chemiluminescence (ECL).

Cheng Z.X., Yin W.B, & Wang Z.Y. (2017). MicroRNA-132 induces temozolomide resistance and promotes the formation of cancer stem cell phenotypes by targeting tumor suppressor candidate 3 in glioblastoma. International Journal of Molecular Medicine, 40(5), 1307-1314.

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CRC Cell Apoptosis Analysis Protocol

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CRC cells were incubated with shDJ-1 for different time periods. Thereafter, they were fixed with 70% icecold ethanol at -20°C for 24 h. Cold PBS was used to wash the cells, which were then cultured with 300 uL of the staining solution (5 U/mL RNaseA and 5 ug/mL PI, BD Pharmingen, USA) for 30 min at 4°C. The results were examined with BD FASCanto II flow cytometry were from subjects with confirmed CRC who were seen at the Second Hospital Affiliated with Nanchang University from 2016 to 2018. The samples were cultured with p-MDM2 (ab170880, Abcam, 1:50), cyclin-D1 (55506T, CST, 1:500), and DJ-1 (ab18257, Abcam, 1:200) antibodies. The staining intensity of the cancer samples was scored as follows: 3 (strong staining, brown), 2 (moderate staining, yellowish-brown), 1 (weak staining, light yellow), and 0 (no staining). Intensity scores < 2 indicated low expression while intensity scores ≥ 2 indicated overexpression. Stained slides were independently examined by two researchers blinded to the clinical outcomes and patient allocation. Metastatic cancer nodules were confirmed with H&E staining.

Zhu X., Luo C., Lin K., Bu F., Ye F., Huang C., Luo H., Huang J, & Zhu Z. (2020). Overexpression of DJ-1 enhances colorectal cancer cell proliferation through the cyclin-D1/MDM2-p53 signaling pathway. Bioscience trends, 14(2).

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