Revision 4

#29603

Store at -20C

Death Receptor Antibody Sampler Kit II

1 Kit

(7 x 20 microliters)

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
Fas (C18C12) Rabbit Monoclonal Antibody423320 µl40-50 kDaRabbit IgG
TNF-R1 (C25C1) Rabbit Monoclonal Antibody373620 µl55 kDaRabbit IgG
TNF-R2 (E8D7P) Rabbit Monoclonal Antibody7233720 µl60-80 kDaRabbit IgG
DR3 (D4O3X) Rabbit Monoclonal Antibody2077220 µl55-60 kDaRabbit IgG
DR4 (D9S1R) Rabbit Monoclonal Antibody4253320 µl35-55 kDaRabbit IgG
DR5 (D4E9) Rabbit Monoclonal Antibody807420 µl40, 48 kDaRabbit IgG
DR6 (E8D2I) Rabbit Monoclonal Antibody9302620 µl80, 120 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

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).

Background References

  1. Nagata, S. (1997) Cell 88, 355-65.
  2. Thorburn, A. (2004) Cell Signal 16, 139-44.
  3. Tartaglia, L.A. et al. (1991) Proc Natl Acad Sci U S A 88, 9292-6.
  4. Peschon, J.J. et al. (1998) J Immunol 160, 943-52.
  5. Tartaglia, L.A. et al. (1993) Cell 73, 213-6.
  6. Rothe, J. et al. (1993) Nature 364, 798-802.
  7. Chinnaiyan, A.M. et al. (1996) Science 274, 990-2.
  8. Kitson, J. et al. (1996) Nature 384, 372-5.
  9. Marsters, S.A. et al. (1996) Curr Biol 6, 1669-76.
  10. Bodmer, J.L. et al. (1997) Immunity 6, 79-88.
  11. Screaton, G.R. et al. (1997) Proc Natl Acad Sci U S A 94, 4615-9.
  12. Marsters, S.A. et al. (1998) Curr Biol 8, 525-8.
  13. Kaptein, A. et al. (2000) FEBS Lett 485, 135-41.
  14. Wang, E.C. et al. (2001) Mol Cell Biol 21, 3451-61.
  15. Osawa, K. et al. (2004) Genes Immun 5, 439-43.
  16. Borysenko, C.W. et al. (2005) Biochem Biophys Res Commun 328, 794-9.
  17. Pan, G. et al. (1997) Science 276, 111-3.
  18. Walczak, H. et al. (1997) EMBO J 16, 5386-97.
  19. Chaudhary, P.M. et al. (1997) Immunity 7, 821-30.
  20. Schneider, P. et al. (1997) Immunity 7, 831-6.
  21. Pan, G. et al. (1998) FEBS Lett 431, 351-6.
  22. Zhao, H. et al. (2001) J Exp Med 194, 1441-8.
  23. Schmidt, C.S. et al. (2003) J Exp Med 197, 51-62.
  24. Nikolaev, A. et al. (2009) Nature 457, 981-9.

Trademarks and Patents

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.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
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.
undefined Image 1: Death Receptor Antibody Sampler Kit II
Western blot analysis of extracts from A549, LN-18 and HeLa cells using TNF-R1 (C25C1) Rabbit mAb.
undefined Image 2: Death Receptor Antibody Sampler Kit II
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.
undefined Image 3: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using DR4 (D9S1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 4: Death Receptor Antibody Sampler Kit II
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.
undefined Image 5: Death Receptor Antibody Sampler Kit II
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.
undefined Image 6: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using DR5 (D4E9) XP® Rabbit mAb.
undefined Image 7: Death Receptor Antibody Sampler Kit II
Western blot analysis of extracts from various cell lines using DR6 (E8D2I) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 8: Death Receptor Antibody Sampler Kit II
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.
undefined Image 9: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Fas (C18C12) Rabbit mAb.
undefined Image 10: Death Receptor Antibody Sampler Kit II
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.
undefined Image 11: Death Receptor Antibody Sampler Kit II
Western blot analysis of extracts from various human cells using TNF-R2 (E8D7P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
undefined Image 12: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
Western blot analysis of extracts from A549 cells, untreated or doxorubicin-treated (500 nM) for the indicated times, using DR5 (D4E9) XP® Rabbit mAb.
undefined Image 13: Death Receptor Antibody Sampler Kit II
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.
undefined Image 14: Death Receptor Antibody Sampler Kit II
Immunohistochemical analysis of paraffn embedded human colon using Fas (C18C12) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
undefined Image 15: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
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.
undefined Image 16: Death Receptor Antibody Sampler Kit II
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.
undefined Image 17: Death Receptor Antibody Sampler Kit II
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.
undefined Image 18: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
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.
undefined Image 19: Death Receptor Antibody Sampler Kit II
Immunohistochemical analysis of paraffin-embedded Raji (positive, left) or K562 cells (negative, right) using Fas (C18C12) Rabbit mAb.
undefined Image 20: Death Receptor Antibody Sampler Kit II
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).
undefined Image 21: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 4

#29603

Death Receptor Antibody Sampler Kit II

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
undefined Image 22: Death Receptor Antibody Sampler Kit II
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.
undefined Image 23: Death Receptor Antibody Sampler Kit II
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.
undefined Image 24: Death Receptor Antibody Sampler Kit II
Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.