Revision 1
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.
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UniProt ID:

#P09874

Entrez-Gene Id:

142

Product Includes Quantity Reactivity MW(kDa) Isotype
Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb 5625 100 µl H Mk 89 Rabbit IgG
PARP (46D11) Rabbit mAb 9532 100 µl H M R Mk 116, 89 Rabbit 

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

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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

PARP, a 116 kDa nuclear poly (ADP-ribose) polymerase, appears to be involved in DNA repair in response to environmental stress (1). This protein can be cleaved by many ICE-like caspases in vitro (2,3) and is one of the main cleavage targets of caspase-3 in vivo (4,5). In human PARP, the cleavage occurs between Asp214 and Gly215, which separates the PARP amino-terminal DNA-binding domain (24 kDa) from the carboxy-terminal catalytic domain (89 kDa) (2,4). PARP helps cells to maintain their viability; cleavage of PARP facilitates cellular disassembly and serves as a marker of cells undergoing apoptosis (6).

  1. Satoh, M.S. and Lindahl, T. (1992) Nature 356, 356-358.
  2. Lazebnik, Y. A. et al. (1994) Nature 371, 346-347.
  3. Cohen, G.M. (1997) Biochem. J. 326, 1-16.
  4. Nicholson, D. W. et al. (1995) Nature 376, 37-43.
  5. Tewari, M. et al. (1995) Cell 81, 801-809.
  6. Oliver, F.J. et al. (1998) J. Biol. Chem. 273, 33533-33539.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus 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 1
    #12061

    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet

    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Etoposide (25 μM, 5 hours) using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb #5625. The virtual lane view (left) shows a single 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.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 2 Expand Image
    Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells treated with Etoposide (25 uM, 5 hours) using PARP (46D11) Rabbit mAb #9532. The virtual lane view (left) shows the target bands (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.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 3 Expand Image
    Enhanced cross-linking and immunoprecipitation (eCLIP) was performed with RNA from K-562 cells and PARP (46D11) Rabbit mAb using a protocol based on the RBP-eCLIP method from Eclipsebio. The figure shows binding across the GET4 transcript. Data is kindly provided by the laboratory of Dr. Gene Yeo and used with permission.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 4 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated or treated with Staurosporine #9953 (1 μM, 3 hr), Jurkat cells, untreated or etoposide-treated (25 μM, overnight), and THP-1 cells, untreated or cycloheximide-treated (CHX, 10 μg/ml, overnight) followed by treatment with TNF-α #8902 (20 ng/ml, 4 hr), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (upper), or total PARP Antibody #9542 (lower).
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 5 Expand Image
    Western blot analysis of extracts from control HEK293 cells (lane 1) or PARP knockout HEK293 cells (lane 2) using PARP (46D11) Rabbit mAb #9532 (upper) or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the PARP knockout HEK293 cells confirms the specificity of the antibody for PARP.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 6 Expand Image
    Immunohistochemical analysis of paraffin-embedded human tonsil using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 7 Expand Image
    Western blot analysis of extracts from THP-1 cells, untreated or treated with TNF-α and cycloheximide as well as control extracts from SW620 and A20 cell lines, using PARP (46D11) Rabbit mAb.
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 8 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (D64E10) XP® Rabbit mAb (green). Actin filament were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    PhosphoPlus® Cleaved PARP (Asp214) Antibody Duet: Image 9 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with Etoposide #2200 (25 uM, 18 hr; green) using Cleaved PARP (Asp214) (D64E10) 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.