Immunofluorescence analysis of PARP was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PARP (Poly ADP-Ribose Polymerase) Rabbit Polyclonal Antibody (Product # PA5-16452) at 2µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjµgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide for the N-terminal region of human PARP|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (Paraffin) (IHC (P))||See 1 publications below|
PA5-16452 targets PARP (Poly ADP-Ribose Polymerase) in IHC (P) and WB applications and shows reactivity with mouse, Rat, and Human samples.
The PA5-16452 immunogen is a synthetic peptide for the N-terminal region of human PARP.
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Generation of Homogenous Three-Dimensional Pancreatic Cancer Cell Spheroids Using an Improved Hanging Drop Technique.
PA5-16452 was used in immunohistochemistry - paraffin section to study the generation of homogenous three-dimensional pancreatic cancer cell spheroids and their improved hanging drop technique
|Ware MJ,Colbert K,Keshishian V,Ho J,Corr SJ,Curley SA,Godin B||Tissue engineering. Part C, Methods (22:312)||2016|