Ab113670

Manufactured by Abcam

Sourced in United States

Ab113670 is a lab equipment product. It is a tool designed for use in research and scientific experiments, but the specific core function is not detailed in an unbiased and factual manner without interpretation or extrapolation.

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Lab products found in correlation

3 protocols using ab113670

Proteomic Analysis of Cell Signaling Molecules

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The hEM15A cells were lysed in RIPA buffer (Sigma, USA) containing protease inhibitor cocktail (Sigma). Then, proteins were isolated using 10% SDS-PAGE gels, transferred onto PVDF membranes (Millipore, USA), and blocked with nonfat milk (5%) at room temperature for 2 h. Afterward, the membranes were incubated with specific primary antibodies against CDK2 (ab32147, 1:1000; Abcam, USA), CDK4 (ab137675, 1:1000), CDK6 (ab124821, 1:1000), Cyclin D1 (ab16663, 1:1000), Bcl-2 (ab32124, 1:1000), Bax (ab32124, 1:1000), cleaved Caspase-3 (ab4051, 1:1000), N-cadherin (ab18203, 1:1000), vimentin (ab137321, 1:1000), E-cadherin (ab15148, 1:1000), TMSB4X (ab167650, 1:1000), or TGF-β2 (ab113670, 1:1000) at 4°C overnight and incubated with corresponding HRP-conjugated secondary antibodies (ab131368 and ab191866; both 1:5,000) for 2 h at room temperature. Eventually, the proteins were visualized via ECL (Thermo Pierce), with GAPDH as the loading control.
Bioinformatics analysis
StarBase website (http://starbase.sysu.edu.cn) was used to find candidate microRNAs (miRNAs) for circPIP5K1A, and miRmap (https://mirmap.ezlab.org), PITA (http://genie.weizmann.ac.il/pubs/mir07/mir07_data.html), and PicTar (http://www.pictar.mdc-berlin.de) databases were used to predict the candidate target genes for miR-153-3p.

Sun L., Wei Y, & Wang J. (2021). Circular RNA PIP5K1A (circPIP5K1A) accelerates endometriosis progression by regulating the miR-153-3p/Thymosin Beta-4 X-Linked (TMSB4X) pathway. Bioengineered, 12(1), 7093-7107.

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Immunoblotting Reagents and Antibodies

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Antibodies against Fli-1 (ab153909, 1:1000), Rb (ab181616, 1:2000), MDM2 (ab38618, 1:1000), Gata-1 (ab181544, 1:1000), Bcl-2 (ab32124, 1:1000), p110 (ab151549, 1:1000), SHIP-1 (ab45142, 1:1000), MMP-1 (ab137332, 1:1000), TGF-β2 (ab113670, 1:1000), phospho-ERK 1/2 (T202 +Y204) (ab223500, 1:1000), and ERK1 (ab32537, 1:1000) were purchased from Abcam (Cambridge, UK). Antibodies against MMP-9 (#13667s, 1:1000), ICAM-1 (#4915s, 1:1000), GAPDH (#2118s, 1:1000), and VEGF-1 (#2893s, 1:1000) were purchased from Cell Signaling Technology (Beverly, MA, USA). Primers (Rb, MDM2, Gata-1, Bcl-2, PI3 K/Ras, SHIP-1, MMP-1, TGF-β2, ERK1, MMP-9, ICAM-1, and VEGF-1) were purchased from GenScript (Nanjing, China). Anti-rabbit and anti-mouse LgG (H+L) [Dylight (TM) 800 4 × PEG Conjugate] secondary antibodies used in this study were purchased from Cell Signaling Technology and used at a 1:30000 dilution in experiments.

Ma Y., Xu B., Yu J., Huang L., Zeng X., Shen X., Ren C., Ben-David Y, & Luo H. (2020). Fli-1 Activation through Targeted Promoter Activity Regulation Using a Novel 3’, 5’-diprenylated Chalcone Inhibits Growth and Metastasis of Prostate Cancer Cells. International Journal of Molecular Sciences, 21(6), 2216.

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Immunofluorescence Analysis of PDR Membranes

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IF staining was performed on 14 PDR membranes, which are seven samples from IVB treatment group, and seven samples from non-treatment group in 10 μm thick frozen sections. We used antibodies against the following antigens: TGFβ2 (1:200 dilution, ab113670, Abcam, Cambridge, Massachusetts, USA), CTGF (1:200 dilution, ab66218, Abcam), CNTF (1:200 dilution, ab66218, Abcam), VEGF (1:200 dilution, ab1316, Abcam) and vimentin (1:150 dilution, Zhongshan Golden Bridge Biotechnology, Beijing, China). Samples were counterstained with DAPI (1:1000 dilution, D9542, Sigma-Aldrich, USA). Sections were examined and photographed using a fluorescence microscope (DS-Ril-U2, Nikon, Tokyo, Japan).

Zhang Q., Qi Y., Chen L., Shi X., Bai Y., Huang L., Yu W., Jiang Y., Zhao M, & Li X. (2016). The relationship between anti-vascular endothelial growth factor and fibrosis in proliferative retinopathy: clinical and laboratory evidence. The British journal of ophthalmology, 100(10).

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