Flip antibody

Manufactured by Adipogen

Sourced in United States

The FLIP antibody is a laboratory reagent that detects the FLIP (Fas-associated death domain-like interleukin-1-beta-converting enzyme-inhibitory protein) protein. FLIP is an important regulator of apoptosis, or programmed cell death. The FLIP antibody can be used in various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and function of FLIP in different biological systems.

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5 protocols using flip antibody

Quantitative Analysis of Apoptosis Markers

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Western blotting was carried out as previously described (21 (link)). cIAP1, cIAP2 and caspase-8 antibodies were from Enzo (Exeter, UK). XIAP, caspase-3, SMAC, APAF-1, caspase-9 and cytochrome c antibodies were from Cell Signaling Technology (Danvers, MA). β-actin antibody was from Sigma (Missouri) and FLIP antibody from AdipoGen (San Diego, CA). Secondary antibodies were purchased from LI-COR Biosciences (Lincoln, NE). Images were captured using an Odyssey Imaging System (LICOR, Lincoln, NE) at 12 bit dynamic range. Quantification of protein expression amounts was conducted as described previously (21 (link)).

Crawford N., Salvucci M., Hellwig C.T., Lincoln F.A., Mooney R.E., O’Connor C.L., Prehn J.H., Longley D.B, & Rehm M. (2018). Simulating and predicting cellular and in vivo responses of colon cancer to combined treatment with chemotherapy and IAP antagonist Birinapant/TL32711. Cell Death and Differentiation, 25(11), 1952-1966.

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Western Blotting and Fractionation

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Western blotting was carried out as previously described^{19 (link)}. Nuclear/Cytoplasmic fractionations are described in detail in Supplementary methods 1. cIAP1- and cIAP2-specific antibodies were from Enzo (Exeter, UK). XIAP, RIPK1, Acetylated-Lysine, HDAC1- and caspase 3-specific antibodies were from Cell Signaling Technology (Danvers, MA, USA). Caspase 8 antibody was from Alexis Biochemicals (San Diego, CA, USA). FLIP antibody was from Adipogen (San Diego, CA, USA). PARP antibody was from eBioscience (San Diego, CA). FADD antibody was obtained from BD Transduction Laboratories (Franklin Lakes, NJ, USA). Ku70 and HSP90 antibodies were from Santa Cruz Biotechnology (Dallas, Texas, USA). Secondary horseradish peroxidase-conjugated antibodies (Amersham, Buckinghamshire, UK) were used for detection on a G-Box digital developer (Syngene. Cambridge, UK).

McCann C., Crawford N., Majkut J., Holohan C., Armstrong C.W., Maxwell P.J., Ong C.W., LaBonte M.J., McDade S.S., Waugh D.J, & Longley D.B. (2018). Cytoplasmic FLIP(S) and nuclear FLIP(L) mediate resistance of castrate-resistant prostate cancer to apoptosis induced by IAP antagonists. Cell Death & Disease, 9(11), 1081.

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Western Blotting for Apoptosis Markers

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Standard Western blotting was carried as described previously (7 (link)). PARP, caspase-8, caspase-3, TRAIL-R2, and Bid antibodies were purchased from Cell Signaling Technology, cIAP1 from Enzo, FADD from BD Biosciences, FLIP antibody from AdipoGen, and β-actin from Sigma. For absolute quantification of FLIP and caspase-8, images were captured using an Odyssey Imaging System (LICOR) at 12-bit dynamic range. Quantification of protein expression amounts was conducted as described previously using recombinant proteins and a HeLa cell line standard (8 (link)).

Grinkevitch V., Wappett M., Crawford N., Price S., Lees A., McCann C., McAllister K., Prehn J., Young J., Bateson J., Gallagher L., Michaut M., Iyer V., Chatzipli A., Barthorpe S., Ciznadija D., Sloma I., Wesa A., Tice D.A., Wessels L., Garnett M., Longley D.B., McDermott U, & McDade S.S. (2022). Functional Genomic Identification of Predictors of Sensitivity and Mechanisms of Resistance to Multivalent Second-Generation TRAIL-R2 Agonists. Molecular Cancer Therapeutics, 21(4), 594-606.

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Western Blotting of Apoptosis Regulators

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Standard Western blotting was carried as previously described (7 (link)). PARP, Caspase-8, Caspase-3, TRAIL-R2 and Bid antibodies were purchased from Cell Signaling Technology, cIAP1 from Enzo, FADD from BD Biosciences, FLIP antibody from AdipoGen and β-actin from Sigma. For absolute quantification of FLIP and Caspase-8, images were captured using an Odyssey Imaging System (LICOR, Lincoln, NE) at 12-bit dynamic range. Quantification of protein expression amounts was conducted as described previously using recombinant proteins and a HeLa cell line standard (8 (link)).

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Quantitative Apoptosis Protein Profiling

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The FEBS Journal 288 (2021) 5374-5388 ª 2021 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies obtained from Enzo (Exeter, UK). FLIP antibody was purchased from AdipoGen (San Diego, CA, USA). RIPK1, RIPK3, XIAP, caspase-3, caspase-9, SMAC, Bid, BCL-2, BCL-XL, MCL-1, Bax, Bak APAF-1 and cytochrome c antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). β-actin antibody was purchased from Sigma (St. Louis, MO, USA). Fluorophore-labelled secondary antibodies were acquired from LI-COR Biosciences (Lincoln, NE, USA). Images were captured using an Odyssey Imaging System (LI-COR) at 12-bit dynamic range. Protein expression in reference cell lines (HeLa/ HT29) was quantified against a standard curve of recombinant protein standards or previously quantified and was used to determine absolute quantities in the cell line panel. Recombinant RIPK1, RIPK3, cIAP1, cIAP2 and XIAP were purchased from Abnova (Taoyuan, Taiwan). p-18 caspase-8 was acquired from Sigma, and FLIP and FADD were produced in-house (PGJCCR, Queen's University Belfast) as described in Majkut et al. [17] . Recombinant Bid, BCL-2, BCL-XL, MCL-1, Bax, Bak, APAF-1, procaspase-9, procaspase-3 and SMAC were used to quantify absolute protein concentrations as previously published [16, 18] .

McCann C., Matveeva A., McAllister K., Van Schaeybroeck S., Sessler T., Fichtner M., Carberry S., Rehm M., Prehn J.H.M, & Longley D.B. (2021). Development of a protein signature to enable clinical positioning of IAP inhibitors in colorectal cancer. The FEBS journal, 288(18).

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