Detection of Human, Mouse, and Rat FLIP by Western Blot.
Western blot shows lysates of PANC‑1 human pancreatic carcinoma cell line, L‑929 mouse fibroblast cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human/Mouse/Rat FLIP Monoclonal Antibody (Catalog # MAB8430) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for FLIP at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
FLIP in Human Pancreatic Cancer Tissue.
FLIP was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Mouse Anti-Human/Mouse/Rat FLIP Monoclonal Antibody (Catalog # MAB8430) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm of cells in islets. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
FLIP (Flice-Like Inhibitory Protein), also known as CFLAR (CASP8 and FADD-Like Apoptosis Regulator), I-FLICE, CASPER, and FLAME-1, is an apoptosis inhibitory protein with architecture similar to that of Caspases-8 and -10. Both of the major FLIP isoforms, the 55 kDa Long (L) and the 27 kDa Short (S), contain two death effector domains (DED). FLIP(L) has a C-terminal Caspase-like protease domain which lacks both a catalytic active site and residues that form a substrate-binding pocket. FLIP(S) and FLIP(L) interact with the adaptor molecule FADD, and potently inhibit apoptosis initiated by Fas Ligand/TNF signaling pathways. Over amino acids 1-200, human FLIP shares 75% sequence identity with mouse and rat FLIP.
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