PA3-007

antibody from Invitrogen Antibodies

Targeting: PDIA5 FLJ30401, PDIR

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA3-007

Provider

Invitrogen Antibodies

Product name

Anti-PDIR Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

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.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

-20° C, Avoid Freeze/Thaw Cycles

References

Submitted references

ERp18 regulates activation of ATF6α during unfolded protein response.

Oka OB, van Lith M, Rudolf J, Tungkum W, Pringle MA, Bulleid NJ
The EMBO journal 2019 Aug 1;38(15):e100990

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot of PDIR in HT 1080 cell lysate using Product # PA3-007.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on membrane enriched extracts (30 µglysate) of U-2 OS (Lane 1), HCT 116 (Lane 2), Hep G2 (Lane 3), A549 (Lane 4), NTERA-2 (Lane 5), HeLa (Lane 6) and Caco-2 (Lane 7). The blot was probed with Anti-PDIR Rabbit Polyclonal Antibody (Product # PA3-007, 1:1,000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 59 kDa band corresponding to PDIR was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

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 (Product # PA3-007) 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.