PARP1 antibody

• Catalog Number: orb420276
• Category: Antibodies
• Description: PARP1 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: 2D-PAGE, IHC, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: PARP1 (N-term ZF1) purified antibody was prepared from whole rabbit serum produced by repeated immunizations with n-terminus region of human PARP1 zinc finger domain recombinant protein.
• Concentration: 1.0 mg/ml
• Dilution range: IHC: 1:100, WB: 1:1000
• Form/Appearance: Liquid (sterile filtered)
• Purity: PARP1 (N-term ZF1) was purified from monospecific antiserum by immunoaffinity chromatography using protein A coupled to agarose beads. This antibody is specific for human PARP1 protein. No cross reactivity detected towards other PARP members when using siRNAs against 18 PARP family members. Cross-reactivity with PARP1 from other sources has not been determined.
• Conjugation: Unconjugated
• UniProt ID: P09874
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-PARP1 Antibody, Poly [ADP-ribose] poly
• Note: For research use only
• Application notes: Anti-PARP1 (N-term ZF1) antibody has been validated by western blotting, IHC, and nanoimmunoassay (NIA). Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 113 kDa in size corresponding to PARP-1 by western blotting in the appropriate cell lysate or extract.
• Expiration Date: 12 months from date of receipt.

Immunohistochemistry with anti-PARP-1 antibody showing nuclear positivity in human lung tissue at 20x and 40x (B & C). Formalin fixed/paraffin embedded tissue sections were subjected to antigen retrieval and then incubated with rabbit anti-PARP-1 antibody orb420276 at 1:100 dilution for 60 minutes. Biotinylated Anti-rabbit secondary antibody was used at 1:200 dilution to detect primary antibody. The reaction was developed using streptavidin-HRP conjugated compact polymer system and visualized with chromogen substrate, 3'3-diamino-benzidine substrate (DAB). The sections were then counterstained with hematoxylin to detect cell nuclei.

OVCAR-8 Wild Type Lysate was separated on 2D SDS-PAGE and blotted on PVDF to analyze immunocoverage of PARP1 antibody specific for the zinc finger 1 domain of PARP1. Primary Antibody: Anti-PARP1 (n-term) antibody 1:200 overnight at 4°C. Secondary Antibody: Goat anti-rabbit Peroxidase (orb347654) at 1:2000 at RT for 30 min. Blocking Buffer: BlockOut (p/n orb348644) for 30 min at RT. Predicted/observed: ~110 kDa and pI 9.7.

Peggy Sue™ Size Separation Electropherogram of OVCAR-8 lysates in no-salt buffer and detected with Anti-PARP1 (N-term ZF1). UV immobilization time: 250 seconds. Protein concentration: 577 µg/mL; 120 s UV immobilization. Primary antibody concentration: 20 µg/mL. Primary antibody incubation time: 180 min. Exposure time: multi-image analysis exposure. Predicted/observed: ~116 kDa.

Western Blot of endogenous PARP1 with Rabbit Anti-PARP1 (N-term ZF1) Antibody. Lane 1: OVCAR8 Wild Type lysate. Lane 2: OVCAR8 PARP1 KO lysate. Load: 5 µg per lane. Primary antibody: PARP1 (N-term ZF1) antibody at 1 µg/mL for overnight at 4°C. Secondary antibody: HRP Gt-a-Rb IgG secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: orb348637 overnight at 4°C. Predicted/Observed size: 113 kDa for endogenous PARP1. Other band(s): none.

Western Blot of endogenous PARP1 with Rabbit Anti-PARP1 Antibodies. Lane 1: OVCAR8 Wild Type lysate. Lane 2: OVCAR8 PARP1 KO lysate. Load 5 µg per lane. Primary Antibody: Blot A: Anti-PARP1- n term (p/n orb420276); Blot B: Anti-PARP1- internal (p/n orb344711) at 1 µg/mL for overnight at 4°C. Secondary antibody: HRP Gt-a-Rb IgG secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: orb348637 overnight at 4°C. Predicted/Observed size: 113 kDa for endogenous PARP1. Other band(s): nonspecific ~ 40 kDa in PARP1-AD only.

Western Blot of Rabbit anti-PARP1 antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate (p/n orb348669). Lane 2: HeLa Lysate (p/n orb348668). Lane 3: MCF-7 Lysate (p/n orb348664). Lane 4: Jurkat Lysate. Lane 5: A431 Lysate (p/n orb348665). Lane 6: A549 Lysate (p/n orb348675). Lane 7: LNCap Lysate (p/n orb348694). Lane 8: MOLT-4 Lysate (p/n orb348696). Lane 9: Ramos Lysate. Lane 10: Raji Lysate (p/n orb348672). Lane 11: A-172 Lysate (p/n orb348708). Lane 12: NIH/3T3 Lysate (p/n orb348714). Load: 35 µg per lane. Primary antibody: PARP1 antibody at 1 ug/ml overnight at 4C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n orb347654) at 1:30000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted/Observed size: 113 kDa for PARP1.

Western Blot of Rabbit anti-PARP1 N-term Antibody. Lane 1: Opal Pre-stained ladder. Lane 2: OVCAR-8 Wild Type. Lane 3: PARP1-KO. Lane 4: PARP2-KO. Lane 5: PARP3-KO. Lane 6: PARP4-KO Lane 7: PARP5a-KO. Lane 8: PARP5b-KO. Lane 9: PARP6-KO. Lane 10: PARP7-KO. Lane 11: PARP8-KO. Lane 12: PARP9-KO. Lane 13: PARP10-KO. Lane 14: PARP12-KO. Lane 15: PARP13-KO. Lane 16: PARP14-KO. Lane 17: PARP16-KO. Load: 5.0 µg per lane. Primary antibody: PARP1 n-term antibody at 1 ug/ml overnight at 4°C. Secondary antibody: Goat anti-rabbit Peroxidase secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Blocking Buffer: orb348644 for 30 min at RT. Predicted/Observed size: ~113 kDa for PARP1.

Western Blot of recombinant PARP1 with rabbit anti-PARP1 (N-term ZF1) antibody. Lane 1: PARP1-Zinc Finger domain recombinant protein. Load: 0.05 µg per lane. Primary antibody: PARP1 (N-term ZF1) antibody at 1 µg/mL for overnight at 4°C. Secondary antibody: HRP Gt-a-rabbit secondary antibody (p/n orb347654) at 1:40000 for 30 min at RT. Block: orb348637 overnight at 4°C. Predicted/Observed size: 13 kDa for rPARP1 (N-term ZF1). Other band(s): none.