Antibody data
- Antibody Data
- Antigen structure
- References [7]
- Comments [0]
- Validations
- Western blot [3]
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- Product number
- 14-6668-82 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), eBioscience™
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Description: This HLNC4 monoclonal antibody reacts with human poly (ADP-ribose) polymerase (PARP1). This ubiquitous 116 kDa nuclear enzyme is involved in DNA repair. During apoptosis, active caspases -3, -6 and -7 cleave PARP1 after Asp214, thereby inactivating PARP1 and generating two apoptotic fragments sized 85 kDa and 25 kDa. The HLNC4 antibody specifically recognizes the 85 kDa PARP1 fragment produced after cleavage and does not recognize the full-length 116 kDa protein.The following peptide was used as the immunogen: NH2-GVDEVAKKKSKKEKDC-COOH. Applications Reported: This HLNC4 antibody has been reported for use in immunoblotting (WB). Applications Tested: This HLNC4 antibody has been tested by immunoblotting of staurosporine-treated Jurkat cells. This can be used at less than or equal to 0.1 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- HLNC4
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- 4° C
Submitted references The G4 Resolvase DHX36 Possesses a Prognosis Significance and Exerts Tumour Suppressing Function Through Multiple Causal Regulations in Non-Small Cell Lung Cancer.
Identification of DHX36 as a tumour suppressor through modulating the activities of the stress-associated proteins and cyclin-dependent kinases in breast cancer.
The role of proteases during apoptosis.
Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase.
Structural and functional analysis of poly(ADP ribose) polymerase: an immunological study.
Production and characterization of monoclonal antibodies specific for the functional domains of poly(ADP-ribose) polymerase.
Poly(ADP-ribose) polymerase is a zinc metalloenzyme.
Cui Y, Li Z, Cao J, Lane J, Birkin E, Dong X, Zhang L, Jiang WG
Frontiers in oncology 2021;11:655757
Frontiers in oncology 2021;11:655757
Identification of DHX36 as a tumour suppressor through modulating the activities of the stress-associated proteins and cyclin-dependent kinases in breast cancer.
Zeng Y, Qin T, Flamini V, Tan C, Zhang X, Cong Y, Birkin E, Jiang WG, Yao H, Cui Y
American journal of cancer research 2020;10(12):4211-4233
American journal of cancer research 2020;10(12):4211-4233
The role of proteases during apoptosis.
Patel T, Gores GJ, Kaufmann SH
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 1996 Apr;10(5):587-97
FASEB journal : official publication of the Federation of American Societies for Experimental Biology 1996 Apr;10(5):587-97
Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase.
Tewari M, Quan LT, O'Rourke K, Desnoyers S, Zeng Z, Beidler DR, Poirier GG, Salvesen GS, Dixit VM
Cell 1995 Jun 2;81(5):801-9
Cell 1995 Jun 2;81(5):801-9
Structural and functional analysis of poly(ADP ribose) polymerase: an immunological study.
Lamarre D, Talbot B, de Murcia G, Laplante C, Leduc Y, Mazen A, Poirier GG
Biochimica et biophysica acta 1988 Jul 13;950(2):147-60
Biochimica et biophysica acta 1988 Jul 13;950(2):147-60
Production and characterization of monoclonal antibodies specific for the functional domains of poly(ADP-ribose) polymerase.
Lamarre D, Talbot B, Leduc Y, Muller S, Poirier G
Biochemistry and cell biology = Biochimie et biologie cellulaire 1986 Apr;64(4):368-76
Biochemistry and cell biology = Biochimie et biologie cellulaire 1986 Apr;64(4):368-76
Poly(ADP-ribose) polymerase is a zinc metalloenzyme.
Zahradka P, Ebisuzaki K
European journal of biochemistry 1984 Aug 1;142(3):503-9
European journal of biochemistry 1984 Aug 1;142(3):503-9
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cell lysates prepared from Jurkat cells left untreated (lane 1) or treated for 2 hrs with staurosporine (lane 2) were immunoblotted with 0.1 µg/mL of the Anti-Human PARP1 (Cleaved) antibody. Bands were visualized using Anti-Mouse IgG HRP.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing of PARP1 (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR978664_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of PARP1 was performed by loading 30 µg of HeLa Wild Type (Lane 1), Treated HeLa Wild Type (Lane 2), HeLa Cas9 (Lane 3), Treated HeLa Cas9 (Lane 4), HeLa Cas9 cells transduced with PARP1 Lentiviral sgRNA (Lane 5) and Treated HeLa Cas9 cells transduced with PARP1 Lentiviral sgRNA (Lane 6) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), eBioscience™ (Product # 14-6668-82) using 1:1,000 dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:4,000 dilution). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Even though NGS analysis determine the clone as partial KO, there was complete loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirming that the antibody is specific to PARP1. Treatment is 1 µM Staurosporin 16 hrs.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PARP1 (cleaved Asp214) Monoclonal Antibody (HLNC4), eBioscience™(Product # 14-6668-82) and a 85 kDa band corresponding to PARP1 (cleaved Asp214) was observed across all cell lines. Nuclear enriched extracts (40 µg lysate) of Jurkat (Lane 1), Jurkat treated with Etoposide (1 µM for 16 hours) (Lane 2), HeLa (Lane 3), HeLa treated with Etoposide (1 µM for 16 hours) (Lane 4), HeLa (Lane 5), HeLa treated with Staurosporine (3 µM for 16 hours) (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.25 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).