Ab61193

Manufactured by Abcam

Sourced in United States

Ab61193 is a primary antibody that can be used for the detection of the target protein in various applications. This product is manufactured by Abcam.

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Lab products found in correlation

4 protocols using ab61193

RIP Assay for AUF1 and Bcl-2 Binding

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RIP was performed using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore, Cambridge, MA). For RIP assay, the supernatant was incubated overnight with beads conjugated with anti-AUF1 (Abcam, ab61193, 1: 50) or anti-Bcl-2 antibody (Abcam, ab32124, 1: 50) or negative control mouse IgG (Millipore). The beads were then rinsed with cold NT2 buffer and cultured with proteinase K at 10 mg/ml (Sigma-Aldrich, USA).

Dai W., Mu L., Cui Y., Li Y., Chen P., Xie H, & Wang X. (2019). Berberine Promotes Apoptosis of Colorectal Cancer via Regulation of the Long Non-Coding RNA (lncRNA) Cancer Susceptibility Candidate 2 (CASC2)/AU-Binding Factor 1 (AUF1)/B-Cell CLL/Lymphoma 2 (Bcl-2) Axis. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 25, 730-738.

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Quantification of EMT-related Protein Levels

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The total proteins were prepared, and their concentrations were determined using a Total Protein Extraction Kit (Solarbio, Beijing, China). Cell lysates were separated by 10% SDS-PAGE and transferred onto nitrocellulose membranes (GE Healthcare, USA). Then, the membranes were blocked with 5% non-fat milk in TBST buffer. The following primary antibodies were used in the study: anti-AUF1 (1:1000, Abcam, ab61193), anti-FOXO3 (1:1000, Abcam, ab17026), anti-E-cadherin (1:1000, Abcam, ab1416), anti-Collagen 1 (1:1000, Abcam, ab6308), anti-α-SMA (1:1000, Abcam, ab5694), and anti-β-actin (1:1000, Abcam, ab8226). Next, the membranes were incubated with HRP-conjugated secondary goat anti-mouse (1:5000, Proteintech, Rosemont, IL, SA00001-1) or goat anti-rabbit (1:5000, Proteintech, SA00001-2) antibodies for 2 h at room temperature. The relative grey values of immunoreactive bands were calculated relative to those of β-actin levels.

Qian W., Cai X., Qian Q., Wang D, & Zhang L. (2020). Angelica Sinensis Polysaccharide Suppresses Epithelial-Mesenchymal Transition and Pulmonary Fibrosis via a DANCR/AUF-1/FOXO3 Regulatory Axis. Aging and Disease, 11(1), 17-30.

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Electrophoretic Mobility Shift Assay of Caco-2 Cells

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EMSA was performed using a LightShift Chemiluminescent EMSA kit (Pierce, Rockford, IL, USA) with nuclear extracts from Caco-2 cells. The mutant and wild-type 29-bp DNA probes (Supplementary Table 3) were synthesized by Integrated DNA Technologies (Coralville, IA, USA). Biotin-labeled DNA was visualized by chemiluminescence. In the supershift assay, for depletion, the following antibodies (1 µl) were added along with the nuclear extract in a 10-µl reaction system: HnRNP K (ab70492, Abcam), hnRNP A/B (sc98810, Santa Cruz), hnRNP Q (ab10687, Abcam), HnRNP D (ab61193, Abcam), and control IgG (ab46540, Abcam).

Xu C., Yang X., Zhou H., Li Y., Xing C., Zhou T., Zhong D., Lian C., Yan M., Chen T., Liao Z., Gao B., Su D., Wang T., Sharma S., Mohan C., Ahituv N., Malik S., Li Q.Z, & Su P. (2019). A novel ZRS variant causes preaxial polydactyly type I by increased sonic hedgehog expression in the developing limb bud. Genetics in Medicine, 22(1), 189-198.

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RNA-Binding Protein Immunoprecipitation Assay

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The RNA immunoprecipitation assay was performed as previously described [16 (link)]. Total RNAs were immunoprecipitated and extracted using an AUF1 antibody (ab61193, Abcam), CPEB1 antibody (ab73287, Abcam), HuD antibody (sc‐28299, Santa Cruz Biotechnology) and normal rabbit IgG (#2729, Cell Signaling Technology, Danvers, MA, USA). cDNA was synthesized from the precipitated RNAs by employing PrimeScript RT Reagent Kit with gDNA Eraser (TaKaRa). qPCR was performed using THUNDERBIRD SYBR qPCR Mix (TOYOBO) and Rotor‐Gene Q Real‐time PCR System (Qiagen).

Oe S., Hayashi S., Tanaka S., Koike T., Hirahara Y., Kakizaki R., Sakamoto S., Noda Y., Yamada H, & Kitada M. (2021). Cpeb1 expression is post‐transcriptionally regulated by AUF1, CPEB1, and microRNAs. FEBS Open Bio, 12(1), 82-94.

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