Anti flip

Manufactured by Enzo Life Sciences

Sourced in Belgium

Anti-FLIP is a laboratory reagent used to detect and quantify the levels of FLIP (Fas-associated death domain-like interleukin-1β-converting enzyme-inhibitory protein) in biological samples. FLIP is a regulator of apoptosis and plays a role in cell survival pathways. Anti-FLIP can be used in various analytical techniques, such as Western blotting, immunoprecipitation, and enzyme-linked immunosorbent assay (ELISA), to measure FLIP expression in cell lines, tissues, or other biological samples.

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Anti-c-FLIP (8 citations) Anti-FLIP antibody (2 citations)

2 protocols using anti flip

Protein Expression Analysis Protocol

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The total cell lysates were prepared by resuspending 0.45×10⁶ cells in 20–50 µl of RIPA lysis buffer (50 mM Tris buffer, 20 mM HEPES, 100 mM NaF; 120 mM NaCl, 0.5% Triton X-100, 100 µM Na₃VO₄, pH 7.6) Total lysates was quantified using a BCA kit (#23225; Thermo Fisher Scientific) according to the manufacturer's protocols. The proteins (30–70 µg) were isolated using 10 or 12% SDS-PAGE gels and electrotransferred onto NC membranes (GE Healthcare). Target proteins were identified using the respective antibodies and Immobilon Western Chemiluminescent HRP Substrate Solution (WBKLS0100; Millipore) and visualized by Davinch-Chemi (CAS-400SM; Davinch-K). Anti-PARP antibody (1:1,000; #9542) and anti-Bax (1:1,000; #2772) were obtain from Cell Signaling Technology. Anti-caspase-3 (1:3,000; ADI-AAP-113) and anti-FLIP (1:700; ALX-804-961-0100) were purchased from Enzo Life Sciences. Anti-Bcl-2 (1:700; sc-7832), anti-Bcl-xL (1:1,000, sc-634), anti-Mcl-1 (1:1,000; sc-12756), anti-cIAP1 (1:1,000; sc-7943), anti-cIAP2 (1:1,000; sc-517317) and anti-β-actin antibody (1:5,000; sc-47778) were supplied by Santa Cruz Biotechnology, and anti-XIAP (1:10,000; 610717) antibody was obtained from BD Biosciences.

Jang J.H., Lee T.J., Sung E.G., Song I.H, & Kim J.Y. (2022). Dapagliflozin induces apoptosis by downregulating cFILPL and increasing cFILPS instability in Caki-1 cells. Oncology Letters, 24(5), 401.

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Comprehensive Histone Enrichment and Western Blot Analysis

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Whole cell extracts were prepared using M-PER^® (Thermofisher, Erembodegen, Belgium) supplemented by 1x protease inhibitor cocktail (Complete EDTA-free, Roche, Prophac, Luxembourg, Luxembourg) according to manufacturer's instructions. Histone enrichment was performed as previously described [14 (link)]. Western blots were performed using the following primary antibodies: anti-DNMT3A (3598), anti-caspase 7 (9494S), anti-caspase 9 (9502S), anti-caspase 8 (9746), anti-PARP (9542), anti-Mcl-1 (4572S), anti-cyclin E1 (4129) from Cell Signaling (Leiden, The Netherlands); anti-DNMT1 (sc-10222), anti-caspase 3 (sc-56053), anti-p21 (sc-817), anti-p27 (sc-527), anti-PCNA (sc-9857R), anti-GRP78 (13968), anti-AHR (8088) from Santa Cruz Biotechnology (Boechout, Belgium); anti-c-myc (51–1485GR), anti-Bcl-xL (610212), anti-XIAP (610763) from BD Pharmigen (Erembodegem, Belgium); anti-FLIP (804–961) from Enzo Life Science, anti-DNMT3B (2851) from Abcam (Cambridge, UK); anti-acetylated histone H4 (06–866), anti-histone H1 (05–457), anti Bcl-2 (OP60) and anti-alpha tubulin (CP06) from Millipore; anti LC-3 (L7543) and anti-beta actin (5441) from Sigma Aldrich; anti-survivin (AF886) from R & D System (Abingdon, UK). Bands were quantified using ImageQuant TL (GE Healthcare, Buckinghamshire, UK) and values of fold change are reported underneath western blots.

Florean C., Schnekenburger M., Lee J.Y., Kim K.R., Mazumder A., Song S., Kim J.M., Grandjenette C., Kim J.G., Yoon A.Y., Dicato M., Kim K.W., Christov C., Han B.W., Proksch P, & Diederich M. (2016). Discovery and characterization of Isofistularin-3, a marine brominated alkaloid, as a new DNA demethylating agent inducing cell cycle arrest and sensitization to TRAIL in cancer cells. Oncotarget, 7(17), 24027-24049.

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