CFLAR/FLIP Polyclonal antibody

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  WB analysis of Jurkat using 10394-1-AP

  Jurkat cells were subjected to SDS PAGE followed by western blot with 10394-1-AP (CFLAR/FLIP antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  WB analysis of HepG2 using 10394-1-AP

  WB result of CFLAR/FLIP antibody (10394-1-AP; 1:500; incubated at room temperature for 1.5 hours) with sh-Control and sh-CFLAR/FLIP transfected HepG2 cells.

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  WB analysis of HeLa using 10394-1-AP

  HeLa cells were subjected to SDS PAGE followed by western blot with 10394-1-AP (CFLAR/FLIP antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours.

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  IP experiment of Jurkat using 10394-1-AP

  IP result of anti-CFLAR/FLIP (IP:10394-1-AP, 4ug; Detection:10394-1-AP 1:500) with Jurkat cells lysate 4000ug.

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  IP experiment of Jurkat using 10394-1-AP

  IP result of anti-CFLAR/FLIP (IP:10394-1-AP, 4ug; Detection:10394-1-AP 1:500) with Jurkat cells lysate 4000ug.

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  IHC staining of human skeletal muscle using 10394-1-AP

  Immunohistochemical analysis of paraffin-embedded human skeletal muscle slide using 10394-1-AP (CFLAR/FLIP antibody at dilution of 1:50.

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  IHC staining of mouse skeletal muscle using 10394-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue slide using 10394-1-AP (CFLAR/FLIP antibody) at dilution of 1:200 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of mouse skeletal muscle using 10394-1-AP

  Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle tissue slide using 10394-1-AP (CFLAR/FLIP antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HeLa using 10394-1-AP

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using CFLAR/FLIP antibody (10394-1-AP) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L), CL594-Phalloidin (red).

• Featured Product
• KD/KO Validated

CFLAR/FLIP Polyclonal Antibody for WB, IHC, IF/ICC, IP, ELISA

Cat No. 10394-1-AP

Publications(10)

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse and More (1)

Applications

WB, IHC, IF/ICC, IP, ELISA

Synonyms

CFLAR, CASP8 and FADD-like apoptosis regulator, CASH, c FLIPS, c FLIPR

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Tested Applications

• Positive WB detected in: Jurkat cells, HepG2 cells, HeLa cells
• Positive IP detected in: Jurkat cells
• Positive IHC detected in: mouse skeletal muscle tissue, human skeletal muscle tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:500-1:2000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunohistochemistry (IHC): IHC : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 9 publications below
• IHC: See 1 publications below

Product Information

10394-1-AP targets CFLAR/FLIP in WB, IHC, IF/ICC, IP, ELISA applications and shows reactivity with human, mouse samples.

• Tested Reactivity: human, mouse
• Cited Reactivity: human, rat
• Host / Isotype: Rabbit / IgG
• Class: Polyclonal
• Type: Antibody
• Immunogen: CFLAR/FLIP fusion protein Ag0593
• Full Name: CASP8 and FADD-like apoptosis regulator
• Calculated Molecular Weight: 52 kDa
• Observed Molecular Weight: 20-35 kDa, 50-55 kDa
• GenBank Accession Number: BC001602
• Gene Symbol: FLIP
• Gene ID (NCBI): 8837
• RRID: AB_2081087
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Antigen affinity purification
• UNIPROT ID: O15519
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

CFLAR, also named CASH, CASP8AP1, CLARP, MRIT, Casper, c-FLIP, and I-FLICE, belongs to the peptidase C14A family. It is an apoptosis regulator protein that may function as a crucial link between cell survival and cell death pathways in mammalian cells. CFLAR acts as an inhibitor of TNFRSF6-mediated apoptosis. It can be cleaved to be 2 fragments P43 and P12. P43 is likely retained in the death-inducing signaling complex (DISC) thereby blocking further recruitment and processing of caspase-8 at the complex. Full-length and shorter isoforms have been shown either to induce apoptosis or to reduce TNFRSF-triggered apoptosis. CFLAR lacks enzymatic (caspase) activity.

Publications

Species	Application	Title
human	WB	Cancer Res; DAB2IP is a bifunctional tumor suppressor that regulates wildtype RAS and inflammatory cascades in KRAS mutant colon cancer; Authors - Abigail L Miller
human	WB	Cell Death Differ; Coordinated repression of BIM and PUMA by Epstein-Barr virus latent genes maintains the survival of Burkitt lymphoma cells.; Authors - Leah Fitzsimmons
rat	WB	J Cell Mol Med; Exercise ameliorates insulin resistance and improves ASK1-mediated insulin signalling in obese rats.; Authors - Yong Zhang
human	WB,IHC	Biomed Pharmacother; Regorafenib suppresses epidermal growth factor receptor signaling-modulated progression of colorectal cancer.; Authors - Yu-Chang Liu
human	WB	Oncol Lett; YM155 sensitizes HeLa cells to TRAIL-mediated apoptosis via cFLIP and survivin downregulation.; Authors - Arun Pandian Chandrasekaran
human	WB	Mol Med Rep; Chidamide induces necroptosis via regulation of c‑FLIPL expression in Jurkat and HUT‑78 cells.; Authors - Zuofei Chi