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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 (PA3-753) at 2ug/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 conjugate (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 (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 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 *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|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"e;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|
antioxidant enzyme AOE372; epididymis secretory sperm binding protein Li 97n; peroxiredoxin IV; Prx 4; prx-IV; thioredoxin peroxidase (antioxidant enzyme); thioredoxin peroxidase AO372; thioredoxin-dependent peroxide reductase A0372
AOE37-2; AOE372; HEL-S-97n; PRDX4; PRX-4