Immunofluorescence analysis of PDIR was performed using 70% confluent log phase U-2 OS 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 PDIR Rabbit Polyclonal Antibody (PA3007) 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 (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 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|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues T(498) N Y I R A L R E G D H E R L G K K(515) of human PDIR.|
|Storage buffer||whole serum|
|Contains||0.01% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:2000|
* 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.
PA3-007 detects PDIR protein in human samples.
PA3-007 has successfully been used in Western blot procedures. By Western blot, this antibody detects an ~58 kDa protein representing PDIR in human fibrosarcoma HT 1080 cell lysate. To a lesser extent, this antibody also detects two unknown, nonspecific bands at 47 and 30 kDa.
The PA3-007 immunogen is a synthetic peptide corresponding to residues T(498) N Y I R A L R E G D H E R L G K K(515) of human PDIR. This peptide (Cat. # PEP-226) is available for use in neutralization and control experiments.
PDIR (protein disulfide isomerase-related protein) is a novel PDI protein that was originally isolated from a human placental cDNA library. It has been suggested that from its nucleotide sequence, PDIR has three CXXC-like motifs (Cys-Ser-Met-Cys, Cys-Gly-His-Cys and Cys-Pro-His-Cys), which are found in the PDI superfamily of proteins. The CXXC motif is responsible for oxidoreductase activity. PDIR is also shown to have a putative endoplasmic reticulum (ER) retention signal 'Lys-Glu-Glu-Leu' at its carboxyl terminus, indicative of ER resident proteins. Northern blot analysis of PDIR indicates that it is preferentially expressed in cells actively secreting proteins. It has also been shown that expression of PDIR is stress-inducible. Data suggest that PDIR, like other PDIs, has oxidoreductase activity of disulfide bonds and facilitates the protein folding process within the lumen of the ER.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.