Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Fas (C18C12) Rabbit mAb.
Western blot analysis of extracts from A549, LN-18 and HeLa cells using TNF-R1 (C25C1) Rabbit mAb.
Western blot analysis of extracts from SW620, NK-92 and SR cell lines, using TNF-R2 Antibody.
Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct encoding full-length human DcR2 (hDcR2; +), using DcR2 (D13H4) Rabbit mAb.
Western blot analysis of lysates from LN18, HeLa and HCT116 cell lines, using DcR3 Antibody.
Western blot analysis of extracts from Jurkat and HeLa cells treated with 4 mM hydroxyurea or 1 µg/ml nocodazole (each for 20 hours), using FADD Antibody (Human Specific).
Western blot analysis of extracts from MCF7, SR, and A673 cell lines, using TRADD (7G8) Rabbit mAb.
Flow cytometric analysis of MCF7 cells using RIP (D94C12) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Immunohistochemical analysis of paraffn embedded human colon using Fas (C18C12) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Western blot analysis of extracts from various cell lines using DR5 (D4E9) XP® Rabbit mAb.
Western blot anlaysis of extracts from various cell lines using DcR2 (D13H4) Rabbit mAb.
Confocal immunofluorescent analysis of OVCAR8 cells using RIP (D94C12) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded Raji (positive, left) or K562 cells (negative, right) using Fas (C18C12) Rabbit mAb.
Western blot analysis of extracts from A549 cells, untreated or doxorubicin-treated (500 nM) for the indicated times, using DR5 (D4E9) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using RIP (D94C12) XP® Rabbit mAb.
Western blot analysis of extracts from COS cells, mock transfected or transfected with human Fas, and from ACHN and HT-1080 cell lines using Fas (C18C12) Rabbit mAb.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with the long isoform of human DR5 (hDR5, +), using DR5 (D4E9) XP® Rabbit mAb.
Western blot analysis of extracts from HeLa cells, untransfected or transfected with human RIP construct, using RIP (D94C12) XP® Rabbit mAb.
|Fas (C18C12) Rabbit mAb 4233||20 µl||
|TNF-R1 (C25C1) Rabbit mAb 3736||20 µl||
|TNF-R2 Antibody 3727||20 µl||
||H M R||65||Rabbit|
|DR5 (D4E9) XP® Rabbit mAb 8074||20 µl||
||H||40, 48||Rabbit IgG|
|DcR2 (D13H4) Rabbit mAb 8049||20 µl||
||H Mk||45-60||Rabbit IgG|
|DcR3 Antibody 4758||20 µl||
|FADD Antibody (Human Specific) 2782||20 µl||
|TRADD (7G8) Rabbit mAb 3684||20 µl||
|RIP (D94C12) XP® Rabbit mAb 3493||20 µl||
||Hm H Mk M R||78||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Death Receptor Antibody Sampler Kit provides an economical means to investigate the machinery of death receptor-mediated apoptosis. The kit includes enough of each primary antibody to perform two western mini-blot experiments per primary.
Each antibody in the Death Receptor Antibody Sampler Kit recognizes endogenous levels of the respective target protein. The TNF-R1, DR5, and RIP antibodies do not cross-react with other related family members. TNF-R1 (C25C1) Rabbit mAb may recognize a 30 kDa splice isoform of TNF-R1 in some cell lines.
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Lys259 of human Fas protein, Ser331 of human TNF-R1 protein, Arg260 of human DR5 protein, Gly227 of human TRADD protein, Leu190 of human RIP protein, or Gly270 of human DcR2 protein.
Polyclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Asp335 of human TNF-R2 protein, Ser194 of human FADD protein, or near the amino terminus of human DcR3 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
The tumor necrosis factor receptor family, which includes TNF-RI, Fas, DR3, DR4, DR5, and DR6, plays an important role in the regulation of apoptosis in various physiological systems (1,2). The receptors are activated by a family of cytokines that include TNF, FasL, and TRAIL. They are characterized by a highly conserved extracellular region containing cysteine-rich repeats and a conserved intracellular region of about 80 amino acids termed the death domain (DD). The DD is important for transducing the death signal by recruiting other DD containing adaptor proteins (FADD, TRADD, RIP) to the death-inducing signaling complex (DISC) resulting in activation of caspases. Death receptor signaling is also controlled by a family of decoy receptors (DcR1, DcR2, and DcR3) which lack a cytoplasmic DD and inhibit death receptor-mediated apoptosis by competing for ligand (3-5). The RIP (receptor-interacting protein) family of serine-threonine kinases (RIP, RIP2, RIP3, and RIP4) are important regulators of cellular stress that can trigger pro-survival and inflammatory responses through the activation of NF-κB as well as pro-apoptotic pathways (6). In addition to the kinase domain, RIP contains a death domain responsible for interaction with the death domain receptor Fas and for the recruitment to TNFR1 through interaction with TRADD (6,7). Overexpression of RIP induces both NF-κB activation and apoptosis (7,8). Caspase-8 dependent cleavage of the death domain on RIP can trigger the apoptotic activity of RIP (9). RIP-deficient cells show a failure in TNF-mediated NF-κB activation, making the cells more sensitive to apoptosis (10,11).
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