Revision 4

#32563

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

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

Applications:
W, IP, IHC-P, IF-IC, FC-FP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
89

Source/Isotype:
Mouse IgG1

UniProt ID:
#P09874

Entrez-Gene Id:
142

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunohistochemistry (Paraffin) 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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.

Specificity/Sensitivity

Cleaved PARP (Asp214) (E2T4K) Mouse mAb recognizes endogenous levels of the large fragment (89 kDa) of human PARP1 protein produced by caspase cleavage. This antibody does not recognize full-length PARP1 or other PARP isoforms.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp214 of human PARP protein.

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).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human

Trademarks and Patents

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

Alexa Fluor is a registered trademark of Life Technologies Corporation.

The transfer of this product is contingent on the buyer using the purchased product solely in research conducted by the buyer (whether the buyer is an academic or for-profit entity), for Immunocytochemistry, high content screening (HCS) analysis, or flow cytometry applications. The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) resale, whether or not such product or its components are resold for use in research; or for any other commercial purpose. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].

This product is provided under an intellectual property license from Life Technologies Corporation. The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or (2) sell or transfer this product or its components for resale, whether or not resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or [email protected].

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

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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

Cell Signaling Technology Logo
Western blot analysis of extracts from serum starved HeLa or HT-29 cells, untreated (-) or treated with Staurosporine #9953 (1 μM; +), using Cleaved PARP (Asp214) (E2T4K) Mouse mAb (upper), total PARP (46D11) Rabbit mAb #9532 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western Blotting Image 1: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
Immunoprecipitation of cleaved PARP (Asp214) from serum-starved HeLa cells treated with Staurosporine #9953 (1 μM, 3 hr). Lane 1 is 10% input, lane 2 is Mouse (G3A1) mAb IgG1 Isotype Control #5415, and lane 3 is Cleaved PARP (Asp214) (E2T4K) Mouse mAb. Western blot was performed using Cleaved PARP (Asp214) (E2T4K) Mouse mAb. Anti-mouse IgG, HRP-linked Antibody #7076 was used as a secondary antibody.
Immunoprecipitation Image 1: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
Immunohistochemical analysis of paraffin-embedded HeLa cell pellet, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemistry Image 1: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
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

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemistry Image 2: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin lymphoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemistry Image 3: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using Cleaved-PARP (Asp214) (E2T4K) Mouse mAb.
Immunohistochemistry Image 4: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
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

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of serum-starved HeLa cells, untreated (left) or treated with Staurosporine #9953 (right), using Cleaved PARP (Asp214) (E2T4K) Mouse mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with etoposide (25 μM, 18 hr; green), using Cleaved PARP1 (Asp214) (E2T4K) Mouse mAb (solid lines) or concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415. Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Flow Cytometry Image 1: Cleaved-PARP (Asp214) (E2T4K) Mouse mAb
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.