Cleaved PARP1 Monoclonal antibody

Cleaved PARP1 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

4G4C8

Cat no : 60555-1-Ig

Synonyms

PARP1, ARTD1, ADPRT1, ADPRT 1, ADPRT

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Validation Data Gallery

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  WB analysis using 60555-1-Ig

  Staurosporine treated and untreated A2780 cells were subjected to SDS PAGE followed by western blot with 60555-1-Ig (Cleaved PARP1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Lamin B1 (HRP-66095) antibody as a loading control.

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  WB analysis using 60555-1-Ig

  Staurosporine treated and untreated mouse splenocytes were subjected to SDS PAGE followed by western blot with 60555-1-Ig (Cleaved PARP1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Lamin B1 (HRP-66095) antibody as a loading control.

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  WB analysis using 60555-1-Ig

  Staurosporine treated and untreated HSC-T6 cells were subjected to SDS PAGE followed by western blot with 60555-1-Ig (Cleaved PARP1 antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Lamin B1 (HRP-66095) antibody as a loading control.

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  IHC staining of Jurkat using 60555-1-Ig

  Immunohistochemical analysis of paraffin-embedded Jurkat (left) and Staurosporine treated Jurkat (right) cells slide using 60555-1-Ig (Cleaved PARP1 antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of HSC-T6 using 60555-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed untreated and 1 μM Staurosporine (3 hours) treated HSC-T6 cells using Cleaved PARP1 antibody (60555-1-Ig, Clone: 4G4C8 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004).

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  IF Staining of HeLa using 60555-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed untreated and 1 μM Staurosporine (3 hours) treated HeLa cells using Cleaved PARP1 antibody (60555-1-Ig, Clone: 4G4C8 ) at dilution of 1:366 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

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  FC experiment of HSC-T6 using 60555-1-Ig

  1x10^6 HSC-T6 cell (dash lines) and 1 μM  Staurosporine (3 hours) treated HSC-T6 cells (full lines) were intracellularly stained with 0.4 μg Cleaved PARP1 Monoclonal Antibody (60555-1-Ig, Clone:4G4C8, red) and CoraLite® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L)(Cat.NO.RGAM005). Mouse IgG1 isotype control (66360-1-Ig, Clone: 1F8D3, blue) was parallel stained as control. Cells were fixed with 4% PFA.

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  FC experiment of HeLa using 60555-1-Ig

  1x10^6 HeLa cell (dash lines) and 1 μM  Staurosporine (3 hours) treated HeLa cells (full lines) were intracellularly stained with 0.1 μg Cleaved PARP1 Monoclonal Antibody (60555-1-Ig, Clone:4G4C8, red) and CoraLite® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L)(Cat.NO.RGAM005). Mouse IgG1 isotype control (66360-1-Ig, Clone: 1F8D3, blue) was parallel stained as control. Cells were fixed with 4% PFA .

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Tested Applications

• Positive WB detected in: A2780 cells, HSC-T6 cells, mouse splenocytes, Staurosporine treated A2780 cells, Staurosporine treated HSC-T6 cells, Staurosporine treated mouse splenocytes
• Positive IHC detected in: Jurkat cells; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: 1 μM Staurosporine (3 hours) treated HSC-T6 cells, 1 μM Staurosporine (3 hours) treated HeLa cells
• Positive FC (Intra) detected in: 1 μM Staurosporine (3 hours) treated HSC-T6 cells, 1 μM Staurosporine (3 hours) treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

60555-1-Ig targets Cleaved PARP1 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: poly (ADP-ribose) polymerase 1
• Calculated Molecular Weight: 1014 aa, 113 kDa
• Observed Molecular Weight: 89 kDa
• GenBank Accession Number: BC037545
• Gene Symbol: PARP1
• Gene ID (NCBI): 142
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

PARP1 (poly(ADP-ribose) polymerase 1) is a nuclear enzyme catalyzing the poly(ADP-ribosyl)ation of many key proteins in vivo. The normal function of PARP1 is the routine repair of DNA damage. Activated by DNA strand breaks, the PARP1 is cleaved into an 85 to 89-kDa COOH-terminal fragment and a 24 kDa NH2-terminal peptide by caspases during the apoptotic process. The appearance of PARP fragments is commonly considered an important biomarker of apoptosis. In addition to caspases, other proteases like calpains, cathepsins, granzymes, and matrix metalloproteinases (MMPs) have also been reported to cleave PARP1 and give rise to fragments ranging from 42-89 kDa.
This antibody only recognizes the cleaved form of PAPR1 but not full-length PARP1.