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8356
Death Receptor Antibody Sampler Kit
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Death Receptor Antibody Sampler Kit #8356

Citations (6)
Simple Western™ analysis of LN18 lysates (1 mg/mL) using TNF-R1 (C25C1) Rabbit mAb #3736. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
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.
Western blot analysis of extracts from HeLa cells, untransfected or transfected with human RIP construct, using RIP (D94C12) XP® Rabbit mAb.
Western blot analysis of extracts from MCF7, SR, and A673 cell lines, using TRADD (7G8) 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 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 lysates from LN18, HeLa and HCT116 cell lines, using DcR3 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.
Flow cytometric analysis of live human peripheral blood mononuclear cells using TNF-R2 (E8D7P) Rabbit mAb (right) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (left), and co-stained with CD14 (61D3) Mouse mAb (APC Conjugate) #36377. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
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 extracts from various cell lines using DR5 (D4E9) XP® 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 human colon carcinoma using Fas (C18C12) Rabbit mAb.
Western blot analysis of extracts from various human cells using TNF-R2 (E8D7P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot anlaysis of extracts from various cell lines using DcR2 (D13H4) 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.
Flow cytometric analysis of control MEF cells (green) or RIP knockout MEF cells (blue) using RIP (D94C12) XP® Rabbit mAb (solid lines) or concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Immunohistochemical analysis of paraffn embedded human colon using Fas (C18C12) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Immunoprecipitation of TNF-R2 protein from SW620 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TNF-R2 (E8D7P) Rabbit mAb. Western blot analysis was performed using TNF-R2 (E8D7P) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.
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.
Simple Western™ analysis of lysates (0.1 mg/mL) from Ramos cells using RIP (D94C12) XP® Rabbit mAb #3493. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (green line) and 1:250 (blue line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Immunohistochemical analysis of paraffin-embedded Raji (positive, left) or K562 cells (negative, right) using Fas (C18C12) Rabbit mAb.
Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
To Purchase # 8356T
Cat. # Size Price Inventory
8356T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Fas (C18C12) Rabbit mAb 4233 20 µl
  • WB
  • IHC
H 40-50 Rabbit IgG
TNF-R1 (C25C1) Rabbit mAb 3736 20 µl
  • WB
  • IP
H 55 Rabbit IgG
TNF-R2 (E8D7P) Rabbit mAb 72337 20 µl
  • WB
  • IP
  • F
H 60-80 Rabbit IgG
DR5 (D4E9) XP® Rabbit mAb 8074 20 µl
  • WB
  • IP
  • IF
H 40, 48 Rabbit IgG
DcR2 (D13H4) Rabbit mAb 8049 20 µl
  • WB
  • IP
H Mk 45-60 Rabbit IgG
DcR3 Antibody 4758 20 µl
  • WB
H R 32 Rabbit 
FADD Antibody 2782 20 µl
  • WB
H 28 Rabbit 
TRADD (7G8) Rabbit mAb 3684 20 µl
  • WB
  • IP
H 32 Rabbit IgG
RIP (D94C12) XP® Rabbit mAb 3493 20 µl
  • WB
  • IP
  • IF
  • F
H M R Hm Mk 78 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

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.

Specificity / Sensitivity

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.

Source / Purification

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. Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human TNF-R2 protein.

Polyclonal antibodies are produced by immunizing animals
with synthetic peptides corresponding to residues surrounding 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.

Background

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).

Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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