Revision 4

#29603Store at -20C

1 Kit

(7 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Fas (C18C12) Rabbit mAb 4233 20 µl 40-50 kDa Rabbit IgG
TNF-R1 (C25C1) Rabbit mAb 3736 20 µl 55 kDa Rabbit IgG
TNF-R2 (E8D7P) Rabbit mAb 72337 20 µl 60-80 kDa Rabbit IgG
DR3 (D4O3X) Rabbit mAb 20772 20 µl 55-60 kDa Rabbit IgG
DR4 (D9S1R) Rabbit mAb 42533 20 µl 35-55 kDa Rabbit IgG
DR5 (D4E9) XP® Rabbit mAb 8074 20 µl 40, 48 kDa Rabbit IgG
DR6 (E8D2I) Rabbit mAb 93026 20 µl 80, 120 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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).

  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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered 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.

    使用に関する制限

    法的な権限を与えられたCSTの担当者が署名した書面によって別途明示的に合意された場合を除き、 CST、その関連会社または代理店が提供する製品には以下の条件が適用されます。お客様が定める条件でここに定められた条件に含まれるものを超えるもの、 または、ここに定められた条件と異なるものは、法的な権限を与えられたCSTの担当者が別途書面にて受諾した場合を除き、拒絶され、 いかなる効力も効果も有しません。

    研究専用 (For Research Use Only) またはこれに類似する表示がされた製品は、 いかなる目的についても FDA または外国もしくは国内のその他の規制機関により承認、認可または許可を受けていません。 お客様は製品を診断もしくは治療目的で使用してはならず、また、製品に表示された内容に違反する方法で使用してはなりません。 CST が販売または使用許諾する製品は、エンドユーザーであるお客様に対し、使途を研究および開発のみに限定して提供されるものです。 診断、予防もしくは治療目的で製品を使用することまたは製品を再販売 (単独であるか他の製品等の一部であるかを問いません) もしくはその他の商業的利用の目的で購入することについては、CST から別途許諾を得る必要があります。 お客様は以下の事項を遵守しなければなりません。(a) CST の製品 (単独であるか他の資材と一緒であるかを問いません) を販売、使用許諾、貸与、寄付もしくはその他の態様で第三者に譲渡したり使用させたりしてはなりません。また、商用の製品を製造するために CST の製品を使用してはなりません。(b) 複製、改変、リバースエンジニアリング、逆コンパイル、 分解または他の方法により製品の構造または技術を解明しようとしてはなりません。また、 CST の製品またはサービスと競合する製品またはサービスを開発する目的で CST の製品を使用してはなりません。(c) CST の製品の商標、商号、ロゴ、特許または著作権に関する通知または表示を除去したり改変したりしてはなりません。(d) CST の製品をCST 製品販売条件(CST’s Product Terms of Sale) および該当する書面のみに従って使用しなければなりません。(e) CST の製品に関連してお客様が使用する第三者の製品またはサービスに関する使用許諾条件、 サービス提供条件またはこれに類する合意事項を遵守しなければなりません。

    Revision 4
    #29603

    Death Receptor Antibody Sampler Kit II

    Death Receptor Antibody Sampler Kit II: Image 1 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 2 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 3 Expand Image
    Western blot analysis of extracts from A549, LN-18 and HeLa cells using TNF-R1 (C25C1) Rabbit mAb.
    Death Receptor Antibody Sampler Kit II: Image 4 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using DR4 (D9S1R) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Death Receptor Antibody Sampler Kit II: Image 6 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 7 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 8 Expand Image
    Western blot analysis of extracts from various cell lines using DR5 (D4E9) XP® Rabbit mAb.
    Death Receptor Antibody Sampler Kit II: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using DR6 (E8D2I) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Death Receptor Antibody Sampler Kit II: Image 10 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 11 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Fas (C18C12) Rabbit mAb.
    Death Receptor Antibody Sampler Kit II: Image 12 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 13 Expand Image
    Western blot analysis of extracts from various human cells using TNF-R2 (E8D7P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    Death Receptor Antibody Sampler Kit II: Image 14 Expand Image
    Western blot analysis of extracts from A549 cells, untreated or doxorubicin-treated (500 nM) for the indicated times, using DR5 (D4E9) XP® Rabbit mAb.
    Death Receptor Antibody Sampler Kit II: Image 15 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 16 Expand Image
    Immunohistochemical analysis of paraffn embedded human colon using Fas (C18C12) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
    Death Receptor Antibody Sampler Kit II: Image 17 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 18 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 19 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 20 Expand Image
    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.
    Death Receptor Antibody Sampler Kit II: Image 21 Expand Image
    Immunohistochemical analysis of paraffin-embedded Raji (positive, left) or K562 cells (negative, right) using Fas (C18C12) Rabbit mAb.
    Death Receptor Antibody Sampler Kit II: Image 22 Expand Image
    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).
    Death Receptor Antibody Sampler Kit II: Image 23 Expand Image
    Confocal immunofluorescent analysis of HT-1080 cells using DR5 (D4E9) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Death Receptor Antibody Sampler Kit II: Image 24 Expand Image
    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.