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Immunofluorescence analysis of PDIA6 (P5) was performed using 70% confluent log phase A-549 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 PDIA6 Rabbit Polyclonal Antibody (PA3-008) 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||Pig|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues D(476) I D L S D V E L D D L G K D E L(492) of human P5.|
|Storage buffer||whole serum|
|Contains||0.01% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|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.
|Western Blot (WB)||See 1 publications below|
PA3-008 detects P5 protein in human samples.
PA3-008 has successfully been used in Western blot procedures. By Western blot, this antibody detects an ~50 kDa protein representing P5 in human fibrosarcoma HT 1080 cell lysate.
The PA3-008 immunogen is a synthetic peptide corresponding to residues D(476) I D L S D V E L D D L G K D E L(492) of human P5. This peptide (Cat. # PEP-224) is available for use in neutralization and control experiments.
P5 is a novel protein that was originally identified in humans. Functionally, P5 has been shown to have peptide binding ability and chaperone activity specific to certain proteins. P5 shares several structural similarities to PDI, an important protein of the ER. The deduced amino acid sequence of P5 shows that it contains the ER retention signal KDEL. P5 also contains two CXXC-like motifs, a motif responsible for oxidoreductase activity. These two CXXC motifs are found on the N- and C-terminus of the protein, one at each end. Studies suggest that the N-terminal CXXC motif is more important than the second for isomerase activity. Contrary to most of the PDI-like proteins, P5 is not stress-inducible.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Protein disulfide isomerase-P5, down-regulated in the final stage of boar epididymal sperm maturation, catalyzes disulfide formation to inhibit protein function in oxidative refolding of reduced denatured lysozyme.
PA3-008 was used in western blot to investigate the physiological roles of PDIA3 and PDI-P5 in mammalian spermiogenesis
|Akama K,Horikoshi T,Sugiyama A,Nakahata S,Akitsu A,Niwa N,Intoh A,Kakui Y,Sugaya M,Takei K,Imaizumi N,Sato T,Matsumoto R,Iwahashi H,Kashiwabara S,Baba T,Nakamura M,Toda T||Biochimica et biophysica acta (1804:1272)||2010|
endoplasmic reticulum protein 5; ER protein 5; protein disulfide isomerase family A, member 6; protein disulfide isomerase P5; protein disulfide isomerase-associated 6; protein disulfide isomerase-related protein; thioredoxin domain containing 7 (protein disulfide isomerase); thioredoxin domain-containing protein 7
ERP5; P5; PDIA6; TXNDC7