- PA3-008 - Invitrogen Antibodies PDIA6 antibody

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

Transmembrane thioredoxin-related protein TMX1 is reversibly oxidized in response to protein accumulation in the endoplasmic reticulum.

Matsuo Y, Hirota K
FEBS open bio 2017 Nov;7(11):1768-1777

Cyclophilin C Participates in the US2-Mediated Degradation of Major Histocompatibility Complex Class I Molecules.

Chapman DC, Stocki P, Williams DB
PloS one 2015;10(12):e0145458

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.

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 2010 Jun;1804(6):1272-84

Multi-chaperone complexes regulate the folding of interferon-gamma in the endoplasmic reticulum.

Vandenbroeck K, Martens E, Alloza I
Cytokine 2006 Mar 7;33(5):264-73

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of A549 (Lane 1), A-431 (Lane 2), HeLa (Lane 3), MCF7 (Lane 4), K-562 (Lane 5), Caco-2 (Lane 6), Hep G2 (Lane 7) and COLO 205 (Lane 8). The blot was probed with Anti-P5 Rabbit Polyclonal Antibody (Product # PA3-008, 1:250 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 48 kDa band corresponding to P5 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).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of PDIA6 was achieved by transfecting HeLa cells with PDIA6 specific siRNAs (Silencer® select Product # S19715). Western blot analysis (Fig a) was performed using membrane enriched extracts from the PDIA6 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti- PDIA6 Rabbit polyclonal Antibody (Product # PA3-008, 1:500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to PDIA6.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 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 (Product # PA3-008) 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of PDIA6 was done on A549 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with PDIA6 Rabbit Polyclonal Antibody (PA3-008, red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 7 Involvement of novel US2-associated proteins in other ERAD pathways (A) US11-mediated degradation. U373-MG + US2 or + US11 cells were treated on day 0 and on day 3 with an siRNA specific for PDIA6 or a negative control. On day 6 cells were lysed and MHC I levels were determined by immunoblotting with mAb HC10. GAPDH served as a loading control. (B) U373-MG, U373-MG + US2, or U373-MG + US11 cells were transiently transfected with either empty pcDNA vector or pcDNA + PDIA6 WT . After 48 h, MHC I levels were determined by immunoblot; BiP served as a loading control. (C) U373-MG cells were treated with the indicated panel of pooled siRNAs on days 0 and 3. On day 4 cells were replated and transiently transfected with an alpha 1 -antitrypsin NHK variant expression plasmid. Approximately 48 h later cells were lysed and NHK levels were determined by immunoblotting.

PA3-008

Antibody data