PARP1 antibody

Catalog No. ABIN967540

Product Details anti-PARP1 Antibody

Target: PARP1 (Poly (ADP-Ribose) Polymerase 1 (PARP1))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This PARP1 antibody is un-conjugated

Application: Western Blotting (WB), Flow Cytometry (FACS), Immunoprecipitation (IP), BioImaging (BI)

Brand: BD Pharmingen™

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human PARP

Clone: 4C10-5

Isotype: IgG1 kappa

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either or Triton™ X-100: a. Add 100 myl of -20°C Triton™ X-100 to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389, ABIN967299

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4°C.

References for anti-PARP1 antibody (ABIN967540)

Ranjit, Cheng, Mackay, Whitacre, Berger, Berger: "Poly(adenosine diphosphoribose) polymerase in peripheral blood leukocytes from normal donors and patients with malignancies." in: Clinical cancer research : an official journal of the American Association for Cancer Research, Vol. 1, Issue 2, pp. 223-34, (1999) (PubMed).

Patel, Gores, Kaufmann: "The role of proteases during apoptosis." in: FASEB journal : official publication of the Federation of American Societies for Experimental Biology, Vol. 10, Issue 5, pp. 587-97, (1996) (PubMed).

Target Details for PARP1

Target: PARP1 (Poly (ADP-Ribose) Polymerase 1 (PARP1))

Alternative Name: PARP (PARP1 Products)

Synonyms: ADPRT antibody, ADPRT 1 antibody, ADPRT1 antibody, ARTD1 antibody, PARP antibody, PARP-1 antibody, PPOL antibody, pADPRT-1 antibody, Adprt antibody, Parp-1 antibody, 5830444G22Rik antibody, AI893648 antibody, Adprp antibody, Adprt1 antibody, C80510 antibody, parp-1 antibody, sPARP-1 antibody, BEST:LD21673 antibody, CG17685 antibody, CG17696 antibody, CG17718 antibody, CG40411 antibody, D.PARP antibody, Dm.pARTa antibody, Dmel\\CG40411 antibody, LD21673.3prime antibody, PARP1 antibody, dPARP antibody, parp antibody, adprt1 antibody, padprt-1 antibody, ppol antibody, si:dkey-206f10.3 antibody, wu:fc60f12 antibody, zgc:110092 antibody, poly(ADP-ribose) polymerase 1 antibody, poly (ADP-ribose) polymerase 1 antibody, poly (ADP-ribose) polymerase family, member 1 antibody, Poly-(ADP-ribose) polymerase antibody, poly(ADP-ribose) polymerase 1 L homeolog antibody, PARP1 antibody, Parp1 antibody, Parp antibody, parp1.L antibody, parp1 antibody

Background: PARP [Poly(ADP-ribose) polymerase] is a 113 kDa nuclear chromatin-associated enzyme that catalyzes the transfer of ADP-ribose units from NAD+ to a variety of nuclear proteins including topoisomerases, histones, and PARP itself. The catalytic activity of PARP is increased in non-apoptotic cells following DNA damage, and PARP is thought to play an important role in mediating the normal cellular response to DNA damage. Additionally, PARP is a target of the caspase protease activity associated with apoptosis. During apoptosis, PARP is cleaved from a 113 kDa intact form into smaller 89 kDa and 24 kDa fragments. This process separates the amino-terminal DNA-binding domain of the enzyme from the carboxy-terminal catalytic domain resulting in the loss of normal PARP function. Although the role of PARP in apoptosis remains to be elucidated, PARP cleavage is considered to be a marker of apoptosis. The 4C10-5 antibody recognizes both the intact 113 kDa form and 89 kDa fragment of PARP.
The 4C10-5 antibody has been reported to recognize both native and denatured PARP. Purified human PARP was used as the immunogen and the antibody reported to react with an epitope located in the NAD binding domain. In dot blot assays, the antibody reacts with the native enzyme in the presence or absence of bound DNA as well as after synthesis of covalently linked poly (ADP-ribose). The 4C10-5 antibody is routinely tested by western blot analysis of untreated Jurkat T cells and Jurkat T cells induced to undergo apoptosis.

Molecular Weight: 113 kD & 89 kD

Pathways: Apoptosis, Caspase Cascade in Apoptosis, DNA Damage Repair, Production of Molecular Mediator of Immune Response, Maintenance of Protein Location