Revision 2

#12894Store at -20C

 

()

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
HAUSP (D17C6) XP® Rabbit mAb 4833 20 µl 135, 140 kDa Rabbit IgG
USP1 (D37B4) Rabbit mAb 8033 20 µl 110 kDa Rabbit IgG
USP2 Antibody 8036 20 µl 68 kDa Rabbit 
USP8 Antibody 8728 20 µl 130 kDa Rabbit 
USP9X Antibody 5751 20 µl 270 kDa Rabbit 
USP10 (D7A5) Rabbit mAb 8501 20 µl 110 kDa Rabbit IgG
USP18 (D4E7) Rabbit mAb 4813 20 µl 34, 39 kDa Rabbit IgG
USP28 Antibody 4217 20 µl 135 kDa Rabbit 
USP14 (D8Q6S) Rabbit mAb 11931 20 µl 60 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 USP Antibody Sampler Kit provides an economical means of detecting members of the ubiquitin-specific protease (USP) family. The kit includes enough primary antibody to perform two western blot experiments per 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

Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme (DUB) action (1,2). The ubiquitin-specific protease (USP) subfamily is one of five distinct groups of DUB enzymes. Ubiquitin-specific-processing protease 1 (USP1) is regulated in a cell cycle dependent manner by both transcriptional and ubiquitin-proteasomal mechanisms (3). Nuclear USP1 localizes to chromatin where it deubiquitinates monoubiquitinated FANCD2 and plays an important role in DNA damage repair and Chk1 protein stability (3,4). Ubiquitin-specific-processing protease 2 (USP2) contains C19 peptidase activity and is involved in ubiquitin recycling and disassembly of polymeric ubiquitin and ubiquitin-like protein complexes (5). USP2 is a putative oncoprotein that is highly over expressed in prostate cancer and drives tumor growth by binding and stabilizing fatty acid synthase through deubiquitination (6,7).

Herpesvirus-associated ubiquitin-specific protease (HAUSP, USP7) binds and deubiquitinates transcription factor p53 and regulator protein Mdm2, stabilizing both proteins (8,9). HAUSP modifies other ubiquitinated proteins, including FoxO family forkhead transcription factors and the mitotic stress checkpoint protein CHFR (10,11). Ubiquitin-specific protease 8 (USP8, UBPy) is a cysteine protease and growth-regulated enzyme that is essential for cell proliferation and survival (12,13). The catalytic domain of USP9X possesses cysteine peptidase activity that cleaves ubiquitin and polyubiquitin conjugates. USP9X may help stabilize adherens and tight junction molecules during epithelial cell polarization (14,15). USP10 is regulated at the posttranslational level through protein-protein interactions and phosphorylation. Interaction of USP10 with the Ras-GAP SH3 domain binding protein (G3BP) inhibits the ability of USP10 to catalyze ubiquitin chain disassembly (16). ATM-mediated phosphorylation of USP10 at Thr42 and Ser337 promotes USP10 stabilization and relocation from the cytoplasm to the nucleus, where it functions in p53 deubiquitination, stabilization, and activation in response to genotoxic stress (17).

USP14 is recruited to the proteasome through association with the PSMD2 (S2/hRPN1) subunit of the 19S regulatory particle, where it may antagonize substrate degradation (18,19). USP14 trims ubiquitin residues from distal polyubiquitin chain ends, decreasing chain affinity for proteasome ubiquitin receptors and allowing for enhanced substrate stability (20,21). USP18 (UBP43) catalyzes the removal of the interferon-regulated, ubiquitin-like protein ISG15 from conjugated proteins (22). Removal of ISG15 from target proteins maintains a critical balance of cellular ISG15-conjugated proteins, which is important for normal development and brain function (23,24). USP28 can bind, deubiquitinate and stabilize several DNA-damage pathway proteins, including p53BP1 and Chk2 (25). USP28 plays an important role in Myc-related signaling as it catalyzes Myc deubiquitination and promotes Myc stabilization, which contributes to tumor-cell growth (26).

  1. Nijman, S.M. et al. (2005) Cell 123, 773-86.
  2. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
  3. Nijman, S.M. et al. (2005) Mol Cell 17, 331-9.
  4. Guervilly, J.H. et al. (2011) Hum Mol Genet 20, 2171-81.
  5. Wilkinson, K.D. (1997) FASEB J 11, 1245-56.
  6. Graner, E. et al. (2004) Cancer Cell 5, 253-61.
  7. Priolo, C. et al. (2006) Cancer Res 66, 8625-32.
  8. Li, M. et al. (2002) Nature 416, 648-53.
  9. Brooks, C.L. et al. (2007) Oncogene 26, 7262-6.
  10. van der Horst, A. et al. (2006) Nat Cell Biol 8, 1064-73.
  11. Oh, Y.M. et al. (2007) Biochem Biophys Res Commun 357, 615-9.
  12. Naviglio, S. et al. (1998) EMBO J 17, 3241-50.
  13. Niendorf, S. et al. (2007) Mol Cell Biol 27, 5029-39.
  14. Murray, R.Z. et al. (2004) Mol Biol Cell 15, 1591-9.
  15. Théard, D. et al. (2010) EMBO J 29, 1499-509.
  16. Soncini, C. et al. (2001) Oncogene 20, 3869-79.
  17. Yuan, J. et al. (2010) Cell 140, 384-96.
  18. Lee, B.H. et al. (2010) Nature 467, 179-84.
  19. Koulich, E. et al. (2008) Mol Biol Cell 19, 1072-82.
  20. Hanna, J. et al. (2006) Cell 127, 99-111.
  21. Thrower, J.S. et al. (2000) EMBO J 19, 94-102.
  22. Malakhov, M.P. et al. (2002) J Biol Chem 277, 9976-81.
  23. Rempel, L.A. et al. (2007) Reprod Biol Endocrinol 5, 13.
  24. Ritchie, K.J. et al. (2002) Genes Dev 16, 2207-12.
  25. Zhang, D. et al. (2006) Cell 126, 529-42.
  26. Popov, N. et al. (2007) Nat Cell Biol 9, 765-74.

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.
    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 2
    #12894

    USP Antibody Sampler Kit

    USP Antibody Sampler Kit: Image 1 Expand Image
    Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and USP10 (D7A5) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the USP10 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
    USP Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various cell lines using USP14 (D8Q6S) Rabbit mAb.
    USP Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from COS cells, untransfected or transfected with mouse USP28, using USP28 Antibody.
    USP Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated or LPS-treated (1 µg/ml, 24 hours), using USP18 (D4E7) Rabbit mAb.
    USP Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from various cell lines using HAUSP (D17C6) XP® Rabbit mAb.
    USP Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using USP9X Antibody.
    USP Antibody Sampler Kit: Image 7 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.
    USP Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from HeLa, A20 and COS cells using USP1 (D37B4) Rabbit mAb.
    USP Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from various cell lines using USP2 Antibody.
    USP Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human USP10 (hUSP10-Myc/DDK, +), using USP10 (D7A5) Rabbit mAb.
    USP Antibody Sampler Kit: Image 11 Expand Image
    Western blot analysis of extracts from various cell lines using USP8 Antibody.
    USP Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing DDK-tagged full-length human USP14 isoform a (hUSP14-DDK; +), using USP14 (D8Q6S) Rabbit mAb.
    USP Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of USP18 from THP-1 extracts treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (80nM overnight) and Lipopolysaccharides (LPS) #14011 (1μg/mL overnight). Lane 1 is 10% input, lane 2 is USP18 (D4E7) Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was perfomed using USP18 (D4E7) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    USP Antibody Sampler Kit: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using HAUSP (D17C6) XP® Rabbit mAb.
    USP Antibody Sampler Kit: Image 15 Expand Image
    Western blot analysis of extracts from COS-7 cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length transcript variant 1 of human USP2 (+), using USP2 Antibody.
    USP Antibody Sampler Kit: Image 16 Expand Image
    Western blot analysis of extracts from various cell lines using USP10 (D7A5) Rabbit mAb.
    USP Antibody Sampler Kit: Image 17 Expand Image
    Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected (+) with a Myc/DDK-tagged cDNA expression construct encoding full-length human USP8, using USP8 Antibody.
    USP Antibody Sampler Kit: Image 18 Expand Image
    Immunoprecipitation of USP14 from 293T cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or USP14 (D8Q6S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using USP14 (D8Q6S) Rabbit mAb.
    USP Antibody Sampler Kit: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse forestomach using HAUSP (D17C6) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
    USP Antibody Sampler Kit: Image 20 Expand Image
    Immunoprecipitation of USP1 protein from COS cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isoype Control #3900, and lane 3 is USP1 (D37B4) Rabbit mAb. Western blot analysis was performed using USP1 (D37B4) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    USP Antibody Sampler Kit: Image 21 Expand Image
    Immunoprecipitation of USP10 from Hep G2 extracts. Lane 1 is 10% input, lane 2 is USP10 (D7A5) Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was perfomed using USP10 (D7A5) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    USP Antibody Sampler Kit: Image 22 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse brain using HAUSP (D17C6) XP® Rabbit mAb.
    USP Antibody Sampler Kit: Image 23 Expand Image
    Confocal immunofluorescent analysis of HCT 116 cells using USP10 (D7A5) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    USP Antibody Sampler Kit: Image 24 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse colon using HAUSP (D17C6) XP® Rabbit mAb.
    USP Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using HAUSP (D17C6) XP® Rabbit mAb.
    USP Antibody Sampler Kit: Image 26 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using HAUSP (D17C6) XP® Rabbit mAb
    USP Antibody Sampler Kit: Image 27 Expand Image
    Confocal immunofluorescent analysis of MCF7 cells using HAUSP (D17C6) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).