PARP1 antibody

• Catalog Number: orb535110
• Category: Antibodies
• Description: PARP1 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: IHC, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: Anti-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. Anti-Rabbit IgG HRP secondary antibody was produced by repeated immunizations in goat with Rabbit IgG whole molecule.
• Form/Appearance: Liquid (sterile filtered)
• Purity: This PARP1 Antibody Set contains: PARP1 (N-term ZF1) protein, Rabbit Anti-PARP1 Antibody, and Goat Anti-RABBIT IgG (HRP) Antibody. PARP1 (N-term ZF1) protein is an N-terminus His-Tag recombinant protein expressed in E.coli corresponding to a fragment of the human PARP1 zinc finger domain. Rabbit Anti-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. Goat Anti-RABBIT IgG (H&L) Antibody Peroxidase Conjugated Pre-Adsorbed was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Peroxidase, anti-Goat Serum, Rabbit IgG and Rabbit Serum. No reaction was observed against or Bovine, Chicken, Goat, Guinea Pig, Hamster, Horse, Human, Mouse, Rat and Sheep Serum Proteins.
• Conjugation: Unconjugated
• UniProt ID: P09874
• Storage: Control Protein and Primary antibody: the vial contains a relatively low volume of reagent (25 µL). Store vial at -20° C or below prior to opening. To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing. Secondary antibody: Store vial at -20° C. For extended storage aliquot contents and freeze at -20° C or below. 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 and 0.01% (w/v) Gentamicin Sulfate
• Alternative names: Poly [ADP-ribose] polymerase 1, ADP-ribosyltransfe
• Note: For research use only
• Application notes: Anti-PARP1 (N-term ZF1) antibody with the matched secondary has been validated by western blotting 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. PARP1 (N-term ZF1) Control Protein analysis by SDS-PAGE resulted in a band at a MW ~13 kDa and estimated to be greater than 95% pure by Coomassie staining.
• 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 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.