Immunofluorescence analysis of Peroxiredoxin 4 was performed using 70% confluent log phase LNCaP 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 PRDX4 Rabbit Polyclonal Antibody (Product # PA3-753) at 1:250 dilution 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 conjugate (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 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified human recombinant Peroxiredoxin 4.|
|Storage buffer||whole serum|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:1000-1:10000|
* 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.
|Western Blot (WB)||See 1 publications below|
PA3-753 detects Peroxiredoxin 4 protein in human samples. This antibody is specific for Prx 4 and shows no cross-reactivity with other Prx isoforms.
PA3-753 has successfully been used in Western blot and immunocytochemistry procedures. By Western blot, this antibody detects an ~23 kDa and ~20 kDa protein representing pro-Prx 4 and mature Prx 4, respectively, in human PC3 cell lysate.
The PA3-753 antigen is purified human recombinant Prx 4 from K562 cells.
The peroxiredoxin (Prdx, Prx, or Trx-Px) family of enzymes is a recently identified family of peroxidases found in free-living organisms. The six known isoforms (Prx1-6) play an important role in protecting lipids, enzymes and DNA against peroxides, such as hydrogen peroxide. The ubiquitously expressed peroxiredoxins have also been shown to play a role in apoptosis and cell differentiation. This is acomplished by the active cysteine of Prx reducing peroxides, which is then converted into a transient cysteine sulfenic acid or cysteine sulfinic acid. If Prx protection is inadequate against peroxidases, the resulting protein and DNA damage may result in neurological disease such as Alzheimer and quote;s or DNA damage leading to cancer.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Nonredundant antioxidant defense by multiple two-cysteine peroxiredoxins in human prostate cancer cells.
PA3-753 was used in western blot to investigate the roles of Prxs in antioxidant defense in human prostate cancer cells
|Shen C,Nathan C||Molecular medicine (Cambridge, Mass.) (8:95)||2002|