co-IP was performed as described previously [17 (link)]. Briefly, the cell lysate was incubated with protein A/G-Sepharose (Novex, Oslo, Norway) and 3 μl antibodies, shook overnight at 4°C, washed with western/IP lysis buffer, and centrifuged at 3500 rpm for 1 minute at room temperature three times. The remaining steps were similar to western blotting. The antibodies used for IP were anti-CREB (CST, #9104), anti-CREB (CST, #9197), anti-ILF2 (Abcam, #ab154169), anti-FLAG (CST, #8146), and anti-HA (CST, #2367). For in vitro reciprocal co-IP, the reagents used were purified protein CREB (Abnova, Taiwan, #H00001385-P01), protein ILF2 (Abnova, #H00003608-P01), and DTT (Beyotime, #ST041) and the method has been described before [18 (link), 19 (link)].
Anti ilf2
Manufactured by Abcam
Sourced in United States
Anti-ILF2 is a protein-specific antibody that recognizes the Interleukin Enhancer Binding Factor 2 (ILF2) protein. ILF2 is a transcriptional regulator involved in various cellular processes. This antibody can be used for research applications involving the detection and study of ILF2 expression in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Dithiothreitol (dtt), by Beyotime (1 mentions)
Protein a g sepharose, by Thermo Fisher Scientific (1 mentions)
Anti ilf3, by Abcam (1 mentions)
Quantity one analysis software, by Bio-Rad (1 mentions)
Immobilon polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
Anti β actin antibody, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
Anti e cadherin, by Abcam (1 mentions)
Anti fibronectin, by Abcam (1 mentions)
Anti vimentin, by Abcam (1 mentions)
Goat anti rabbit igg h l hrp secondary antibody, by Abcam (1 mentions)
Anti n cadherin, by Abcam (1 mentions)
Anti β actin, by Abcam (1 mentions)
Odyssey infrared imaging system, by LI COR (1 mentions)
3 protocols using anti ilf2
1
CREB, ILF2 Protein Interaction Analysis
2
Western Blot Analysis of ILF2, ILF3, EEF1A1, and Rbmxl1
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Cells or tissues were harvested at the indicated times and lysed in radioimmunoprecipitation assay (RIPA) extraction solution (15 mM Tris, pH 7.5, 120 mM NaCl, 25 mM KCl, 2 mM EGTA, 2 mM EDTA, 0.1 mM dithiothreitol, 0.5% Triton X-100, and protease inhibitor cocktail [Sigma]). The protein concentration was assessed by bicinchoninic acid (BCA) assay. Total protein lysates (25 μg/lane) were subjected to SDS-PAGE and transferred onto an Immobilon polyvinylidene difluoride (PVDF) membrane (Millipore). The antibodies used were the following: anti-ILF2 (Abcam), anti-ILF3 (Abcam), anti-EEF1A1 (Novus), anti-Rbmxl1 (Novus), and anti-β-actin antibody as the loading control (Santa Cruz Biotechnology). Western blots were quantified by Quantity One Analysis software (Bio-Rad).
3
Western Blot Analysis of EMT Markers
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Western blot analysis was performed as previously described (18 (link)). In brief, after incubation with the primary antibodies, anti-ILF2 (Cat no. ab154791; 1:250), anti-E-cadherin (Cat no. ab40772; 1:250), anti-N-cadherin (Cat no. ab18203; 1:250), anti-vimentin (Cat no. ab45939; 1:250), anti-fibronectin (Cat no. ab2413; 1:250) and anti-β-actin (Cat no. ab8227; 1:500) (all from Abcam, Cambridge, MA, USA) overnight at 4°C, goat anti-rabbit IgG H&L (HRP) secondary antibody (Cat no. ab7090; Abcam) was used for 30 min at room temperature. The specific proteins were visualized using the Odyssey™ infrared imaging system (Li-COR, Biosciences).
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