MA1-41020

antibody from Invitrogen Antibodies

Targeting: PARP1 ADPRT, ARTD1, PARP, PPOL

Provider product page for MA1-41020

Western blot

Immunocytochemistry

Flow cytometry

Antibody data

Antibody Data
Antigen structure
References [0]
Comments [0]
Validations
Western blot [4]
Immunocytochemistry [1]

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Product number: MA1-41020 - Provider product page
Provider: Invitrogen Antibodies
Product name: PARP1 (cleaved Asp214, Asp215) Monoclonal Antibody (194C1439)
Antibody type: Monoclonal
Antigen: Synthetic peptide
Description: Suggested positive control: camptothecin treated HL-60 (12 hr).
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: 194C1439
Vial size: 100 µg
Concentration: 1.0 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

PARP1 protein structure - MA1-41020 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of cleaved PARP in staurosporine-treated Jurkat cells at various time points, using a PARP monoclonal antibody (Product # MA1-41020) at 2 µg/mL. The band corresponding to cleaved PARP is only seen in the treated samples.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of PARP1 (cleaved Asp214, Asp215) in staurosporine-treated Jurkat cells. Samples were incubated in PARP1 (cleaved Asp214, Asp215) monoclonal antibody (Product # MA1-41020) using a dilution of 2 µg/mL followed by an Anti-mouse Ig HRP conjugate secondary antibody. The band corresponding to cleaved PARP is only seen in the treated samples.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-PARP Monoclonal Antibody (194C1439) (Product # MA1-41020) and a 85 kDa band corresponding to PARP (cleaved form) was observed across all cell lines along with an uncharacterized band (*) at ~50 kDa. 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 (1 µg/mL concentration) 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

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, Asp215) Monoclonal Antibody (194C1439) (Product # MA1-41020) 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 confirming that the antibody is specific to PARP1. An uncharacterized band was observed at 49 kDa in HeLa Wild Type and Cas9 untreated samples. Treatment