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.
Western blot analysis of various cell lines using DR3 (D4O3X) Rabbit mAb (upper), or β-Actin (D6A8) Rabbit mAb #8457 (lower). KARPAS cell line source: Dr. Abraham Kapas at the University of Cambridge.
Flow cytometric analysis of fixed and permeabilized U266 cells using DR4 (D9S1R) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunoprecipitation of DR6 from RT4 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DR6 (E8D2I) Rabbit mAb. Western blot was performed using DR6 (E8D2I) Rabbit mAb. Anti-Rabbit, HRP-linked Antibody #7074 was used as a secondary antibody.
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 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full length human DR3 (hDR3-Myc/DDK; +) using DR3 (D4O3X) Rabbit mAb.
Confocal immunofluorescent analysis of HCT 116 (positive, left) and FaDu (negative, right) cells using DR4 (D9S1R) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Western blot analysis of extracts from various cell lines using DR5 (D4E9) XP® Rabbit mAb.
Western blot analysis of extracts from various cell lines using DR6 (E8D2I) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemical analysis of paraffin-embedded Raji (positive, left) or K562 cells (negative, right) using Fas (C18C12) Rabbit mAb.
Immunoprecipitation of DR4 from PC-3 cell extracts. Lane 1 represents 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is DR4 (D9S1R) Rabbit mAb. Western blot was performed using DR4 (D9S1R) Rabbit mAb. A conformation specfic secondary antibody was used to avoid IgG cross-reactivity.
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 293T cells, mock transfected (-) or transfected with a construct expressing full-length human DR6 (hDR6; +), using DR6 (E8D2I) 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 various cell lines using DR4 (D9S1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
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 293T cells, mock transfected (-) or transfected with a construct expressing full-length human DR4 (hDR4; +) using DR4 (D9S1R) 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|
|DR3 (D4O3X) Rabbit mAb 20772||20 µl||
|DR4 (D9S1R) Rabbit mAb 42533||20 µl||
|DR5 (D4E9) XP® Rabbit mAb 8074||20 µl||
||H||40, 48||Rabbit IgG|
|DR6 (E8D2I) Rabbit mAb 93026||20 µl||
||H||80, 120||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Death Receptor Antibody Sampler Kit II provides an economical means to investigate members of the death receptor family. The kit includes enough antibody to perform two western blot experiments with each primary antibody.
Each antibody in the Death Receptor Antibody Sampler Kit II detects endogenous levels of its target protein. TNF-R1 (C25C1) Rabbit mAb recognizes a 30 kDa splice isoform of TNF-R1 in some cell lines. DR3 (D4O3X) Rabbit mAb detects a band unknown origin at 110 kDa in some cell lines.
Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Lys259 of human Fas, Ser331 of human TNF-R1, Leu335 of human DR3, Arg260 of human DR5, His235 of human DR6, or a recombinant protein within the carboxy terminus of DR4. The polyclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp339 of human TNF-R2. Antibodies are purified by protein A and peptide affinity chromatography.
The tumor necrosis factor receptor family, which includes TNF-RI, TNF-R2, 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, TWEAK, 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. The two receptors for TNF-α, TNF-R1 (55 kDa) and TNF-R2 (75 kDa) can mediate distinct cellular responses (3,4). In most cases cytotoxicity elicited by TNF has been reported to act through TNF-R1 (5,6). DR3/WSL-1/Apo-3/TRAMP/LARD is a TNFR family member containing the characteristic extracellular cysteine-repeats, transmembrane region, and an intracellular DD (7-11). DR3 is activated by its ligand Apo-3L/TWEAK to induce apoptosis and activation of NF-κB (12,13). Like TNF-R1, DR3 binds to the DD adaptor protein TRADD, which can then associate with other DD proteins like FADD and RIP as well as members of the TRAF family (7,8). Tissue expression of DR3 is very restricted, primarily seen on the surface of activated thymocytes and lymphocytes and plays an important role in thymocyte negative selection (7,8,14). Studies have also indicated an association with DR3 and rheumatoid arthritis (15,16). DR4 (TRAIL-RI, TNFRSF10A) and DR5 (TRAIL-R2, TNFRSF10B) are receptors for the cytokine TRAIL. Both receptors contain death domains that recruit DISC complexes triggering caspase activation and apoptosis (17-20). DR6, also known as TNFRSF21, is a TNFR family member able to induce apoptosis as well as activation of NF-κB and JNK (21). DR6 appears to play a critical role in the activation and differentiation of T and B lymphocytes (22,23). In the nervous system, β-amyloid precursor protein (APP) activates DR6 to trigger neuronal degeneration (24).
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