Invitrogen™ Human PARP (Cleaved) [214/215] ELISA Kit

Description

ELISA

The Human PARP (Cleaved) [214/215] ELISA Kit is designed to detect and quantify the level of PARP (Cleaved) [214/215] ) in fresh or frozen human cell lysates. The assay recognizes both natural and recombinant human PARP (Cleaved) [214/215]. Principle of the method A monoclonal capture antibody specific for PARP (Cleaved) [214/215] has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Poly ADP-Ribose Polymerase (PARP) uses nicotinamide adenine dinucleotide (oxidized form) NAD as a substrate to catalyse the transfer of ADP-ribose to a variety of nuclear protein acceptors. Proteolysis of PARP to its stable 85kDa fragment is an early marker of programmed cell death (apoptosis) and is mediated by the caspase CPP32 protein. Cleavage occurs between Adp216 and Gly217, a site in PARP conserved across species.

Specifications

• Accession Number: P09874
• Assay Range: 0.156 to 10 ng/mL
• Assay Sensitivity: <0.062 ng/mL
• Concentration: 0.156 to 10 ng/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Cell Lysates
• Target Species: Human
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 142
• Gene Symbol: ADPRT,ADPRT 1,ADPRT1,ARTD1,PARP,PARP-1,PPOL,pADPRT-1
• Incubation Time: 4 hr.
• Interassay CV: 5.3%
• Intraassay CV: 4.6
• Kit Contents: Pre-coated 96 well plate, Standard, Standard Dilution Buffer, Biotinylated Detection Antibody, Streptavidin-HRP, HRP Diluent, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Marker Gene: PARP
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate 10 μL
• Storage Requirements: 2°C to 8°C
• Target: PARP1 (cleaved Asp214)
• Test Time: 1 hrs 20 mins
• Total Assay Time: 4 hrs