Protein Disulfide Isomerase/P4HB Antibody (RL90)

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of Protein Disulfide Isomerase/P4HB was performed by loading 25 ug of HepG2 (Lane 1), Hela (Lane 2) and NIH-3T3 (Lane 3) cell lysates.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining of Phalloidin (purple) and PDI (green) in NIH 3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 650 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Both normal and cancer biopsies of deparaffinized Human colon tissue tissues.

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of Hela cells compared to an isotype control (blue).

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of PDIA2 expression in 1) human brain and 2) rat liver tissues extracts, 3) HeLa and 4) Hek 293 whole cell lysates.

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cell surface Protein Disulfide Isomerase/P4HB associated with GC-AC. Proteins were pulled down with anti GC-A antibody from HUVEC or HMC membrane preparations and detected with anti-Protein Disulfide Isomerase/P4HB antibody (RL90)compared to control IP (no anti GC-A) and non-immunoprecipitated proteins from HMC and HUVEC. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in p19 Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with Dylight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with Dylight 350 Phalloidin at a dilution of 1:120 (2.5units/ml final concentration) and nuclei (red) were stained with DRAQ5 at a concentration of 1ug/ml for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 550 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 488 Phalloidin at a dilution of 1:300 (1 unit/ml final concentration) for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Formalin fixed cells were permeabilized with 0.1% Triton X-100 in PBS for 10 minutes at room temperature and blocked with 2% BSA in PBS + 0.1% Triton X-100 for 30 minutes at room temperature. Cells were probed with a Protein Disulfide Isomerase/P4HB monoclonal antibody at a dilution of 1:75 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody at a dilution of 1:250 for 30 minutes at room temperature. Actin was stained with DyLight 550 Phalloidin at a dilution of 1:120 (2.5 units/ml final concentration) and nuclei (blue) were stained with Hoechst at a concentration of 1ug/ml for 30 minutes.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in A549 Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in HMVEC Cells.

Immunocytochemistry/Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Staining in NS-1 Cells.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cancer biopsies of deparaffinized Human lung adenocarcinoma tissues.

Immunohistochemistry-Paraffin: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Both normal and cancer biopsies of deparaffinized Human tonsil tissues.

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of NIH/3T3 cells compared to an isotype control (blue).

Flow Cytometry: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Analysis of K562 cells compared to an isotype control (blue).

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Down regulated PDIA2 attenuates CNP-mediated cGMP activation in HMCs. Western blotting with RL90 antibody shows partial knockdown of Protein Disulfide Isomerase/P4HB in HMCs.

Immunocytochemistry/ Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cell surface PDI associated with GC-A & GC-B.A. & C. Proteins were pulled down with anti GC-A or anti GC-B antibodies from HUVEC or HMC membrane preparations & detected with anti-PDI antibody (RL90)compared to control IP (no anti GC-A or anti GC-B antibody) & non-immunoprecipitated proteins from HMC & HUVEC. B. Co-localization of PDI (red) & GC-A (green) in HUVEC. D. Co-localization of PDI (green) & GC-B (red) in HMC. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Down regulated PDI attenuates CNP-mediated cGMP activation in HMCs.A. Western blotting with RL90 antibody shows partial knockdown of PDI in HMCs. B. Effect of PDI knockdown on cGMP activation. Data are expressed as mean+SEM vs control cells without PDI siRNA treatment (*P≤0.05). C. PDI inhibition by RL90 decreases ANP, BNP & CNP-mediated cGMP generation in HMC (*p≤0.05). (Error bars, +SD from 3 independent experiments, samples were triplicated in each experiment). Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cell surface PDI associated with GC-A & GC-B.A. & C. Proteins were pulled down with anti GC-A or anti GC-B antibodies from HUVEC or HMC membrane preparations & detected with anti-PDI antibody (RL90)compared to control IP (no anti GC-A or anti GC-B antibody) & non-immunoprecipitated proteins from HMC & HUVEC. B. Co-localization of PDI (red) & GC-A (green) in HUVEC. D. Co-localization of PDI (green) & GC-B (red) in HMC. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: Protein Disulfide Isomerase/P4HB Antibody (RL90) [NB300-517] - Cell surface PDI associated with GC-A & GC-B.A. & C. Proteins were pulled down with anti GC-A or anti GC-B antibodies from HUVEC or HMC membrane preparations & detected with anti-PDI antibody (RL90)compared to control IP (no anti GC-A or anti GC-B antibody) & non-immunoprecipitated proteins from HMC & HUVEC. B. Co-localization of PDI (red) & GC-A (green) in HUVEC. D. Co-localization of PDI (green) & GC-B (red) in HMC. Image collected & cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0112986), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ha, Pm, Pm, Rb
• Applications: WB, ELISA, Flow, ICC/IF, IHC, IP, B/N, KD
• Clone: RL90
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

Protein Disulfide Isomerase/P4HB Antibody (RL90) Summary

• Immunogen: Purified rat Protein Disulfide Isomerase/P4HB protein.
• Marker: Endoplasmic Reticulum Marker
• Specificity: NB300-517 detects Protein Disulfide Isomerase/P4HB from human, rat, porcine and mouse tissues as well as hamster cells.
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: P4HB
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • Block/Neutralize
  • ELISA 1:100 - 1:2000
  • Flow Cytometry 1:10 - 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry 1:10 - 1:500
  • Immunohistochemistry-Frozen 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Immunomicroscopy 1:10 - 1:500
  • Immunoprecipitation 1:10 - 1:500
  • Knockdown Validated
  • Western Blot 1:1000
• Application Notes: IHC usage was reported in scientific literature (see Tanaka S, et al). ELISA assay reported in literature (see Kaetzel CS, et al). Flow Cytometry usage was reported in scientific literature (see Ryser HJ, et al). Blocking/Neutralization usage was reported in the literature (PMID:12791669 and PMID: 22013897)
• Control:

  HeLa Whole Cell Lysate

Reactivity Notes

Rabbit reactivity reported in scientific literature (PMID: 16150729). Bovine (PMID: 14592831). Primate (PMID: 20667829). Monkey reactivity reported in scientific literature (PMID: 27487388). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: PBS with 1 mg/ml BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Protein A purified

Alternate Names for Protein Disulfide Isomerase/P4HB Antibody (RL90)

• Cellular thyroid hormone-binding protein
• collagen prolyl 4-hydroxylase beta
• DSI
• ERBA2L
• GIT
• glutathione-insulin transhydrogenase
• P4HB
• P4Hbeta
• p55
• PDI
• PDIA1
• PDIA1procollagen-proline, 2-oxoglutarate 4-dioxygenase (proline 4-hydroxylase), betapolypeptide (protein disulfide isomerase-associated 1)
• PDIEC 5.3.4.1
• PHDB
• PO4DB
• PO4HB
• procollagen-proline, 2-oxoglutarate 4-dioxygenase (proline 4-hydroxylase), betapolypeptide
• PROHB
• PROHBprocollagen-proline, 2-oxoglutarate 4-dioxygenase (proline 4-hydroxylase), betapolypeptide (protein disulfide isomerase; thyroid hormone binding protein p55)
• Prolyl 4-hydroxylase subunit beta
• prolyl 4-hydroxylase, beta polypeptide
• protein disulfide isomerase family A, member 1
• protein disulfide isomerase/oxidoreductase
• protein disulfide isomerase-associated 1
• protein disulfide-isomerase
• protocollagen hydroxylase
• Thbp
• thyroid hormone-binding protein p55

Background

P4HB, the beta subunit of prolyl 4-hydroxylase, a highly abundant member of the PDI family. P4HB is a multifunctional enzyme, involved in hydroxylation of prolyl residues in preprocollagen, as well as continual rearrangement of disulfide bonds. Other known functions include its ability to act as a chaperone that inhibits aggregation of misfolded proteins in a concentration-dependent manner, its ability to bind thyroid hormone, its role in both the influx and efflux of S-nitrosothiol-bound nitric oxide, and its function as a subunit of the microsomal triglyceride transfer protein complex.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Protein Disulfide Isomerase/P4HB Antibody (NB300-517)(28)

Publications using NB300-517

• Voronkova MA, Johnson B, Gandhi N et al. ERO1A levels are a prognostic indicator in EGFR mutated non small cell lung cancer NPJ Precision Oncology 2024-11-04 [PMID: 39496753]
• Stempels FC, Jiang M, Warner HM et al. Giant worm-shaped ESCRT scaffolds surround actin-independent integrin clusters The Journal of cell biology 2023-07-03 [PMID: 37200023] (ICC/IF, Human)
• Li Y, Kubo H, Yu D et al. Combining three independent pathological stressors induces a heart failure with preserved ejection fraction phenotype American journal of physiology. Heart and circulatory physiology 2023-04-01 [PMID: 36763506] (IHC-P, Mouse)
  
  Details:
  1:100 dilution
• Lo M, Sharir A, Paul MD et al. CNPY4 inhibits the Hedgehog pathway by modulating membrane sterol lipids Nature communications 2022-05-03 [PMID: 35504891] (ICC/IF, Human)
  
  Details:
  Fig. 5a
• Avinash Kumar, Nicole Welch, Saurabh Mishra, Annette Bellar, Rafaella Nasciemento Silva, Ling Li, Shashi Shekhar Singh, Mary Sharkoff, Alexis Kerr, Aruna Kumar Chelluboyina, Jinendiran Sekar, Amy H. Attaway, Charles Hoppel, Belinda Willard, Gangarao Davuluri, Srinivasan Dasarathy Metabolic reprogramming during hyperammonemia targets mitochondrial function and postmitotic senescence JCI Insight 2021-12-22 [PMID: 34935641]
• Peter T. A. Linders, Eveline C. F. Gerretsen, Angel Ashikov, Mari-Anne Vals, Rinse de Boer, Natalia H. Revelo, Richard Arts, Melissa Baerenfaenger, Fokje Zijlstra, Karin Huijben, Kimiyo Raymond, Kai Muru, Olga Fjodorova, Sander Pajusalu, Katrin Õunap, Martin ter Beest, Dirk Lefeber, Geert van den Bogaart Congenital disorder of glycosylation caused by starting site-specific variant in syntaxin-5 Nature Communications 2021-10-28 [PMID: 34711829]
• Beckmann L, Rolling CC, VoigtlAnder M Et al. Bacitracin and Rutin Regulate Tissue Factor Production in Inflammatory Monocytes and Acute Myeloid Leukemia Blasts Cancers 2021-08-04 [PMID: 34439096]
• Paardekooper LM, van Vroonhoven E, Ter Beest M, van den Bogaart G Radical Stress Is More Cytotoxic in the Nucleus than in Other Organelles Int J Mol Sci 2019-08-25 [PMID: 31450682] (WB, Human)
• Subramanian A, Capalbo A, Iyengar NR et al. Auto-regulation of Secretory Flux by Sensing and Responding to the Folded Cargo Protein Load in the Endoplasmic Reticulum Cell 2019-03-07 [PMID: 30849374] (ICC/IF, Human)
• Liang D, Wu Y, Zhou L et al. LRP5 controls cardiac QT interval by modulating the metabolic homeostasis of L-type calcium channel. Int J Cardiol. [PMID: 30309679] (ICC/IF, Mouse)
• Zorin VL, Pulin AA, Eremin II et al. Myogenic potential of human alveolar mucosa derived cells Cell Cycle 2017-03-19 [PMID: 28118065] (ICC/IF, FLOW, Human)
• Dosoki H, Stegemann A, Taha M et al. Targeting of NADPH oxidase in vitro and in vivo suppresses fibroblast activation and experimental skin fibrosis. Exp. Dermatol. 2016-08-30 [PMID: 27576129]
• Walsh MT, Di Leo E, Okur I et al. Structure-function analyses of microsomal triglyceride transfer protein missense mutations in abetalipoproteinemia and hypobetalipoproteinemia subjects. Biochim. Biophys. Acta. 2016-07-31 [PMID: 27487388] (WB, Monkey)
• Pan S, Chen Hh, Correia C et al. Cell surface protein disulfide isomerase regulates natriuretic Peptide generation of cyclic guanosine monophosphate. PLoS OnE. 2014-11-25 [PMID: 25419565] (ICC/IF, Human)
• Langer F, Spath B, Fischer C et al. Rapid activation of monocyte tissue factor by antithymocyte globulin is dependent on complement and protein disulfide isomerase. Blood 2013-01-11 [PMID: 23315166]
• Hicks DA, Makova NZ, Nalivaeva NN et al. Characterisation of acetylcholinesterase release from neuronal cells. Chem Biol Interact. 2013-03-25 [PMID: 23047022] (B/N, Mouse)
• Elahi S, Niki T, Hirashima M et al. Galectin-9 binding to Tim-3 renders activated human CD4+ T cells less susceptible to HIV-1 infection. Blood. 2012-05-03 [PMID: 22438246] (B/N, Human)
• Vallon M, Aubele P, Janssen K-P, Essler M. Thrombin-induced shedding of tumor endothelial marker 5 and exposure of its RGD motif are regulated by cell surface protein disulfide isomerase. The Biochemical Journal. 2011-10-20 [PMID: 22013897]
• Paule S, Li Y, Nie G. Cytoskeletal remodelling proteins identified in fetal-maternal interface in pregnant women and rhesus monkeys. J Mol Histol;42(2):161-6. 2011-04-01 [PMID: 21373848] (IF/IHC, Primate)
• Paule SG, Airey LM, Li Y et al. Proteomic approach identifies alterations in cytoskeletal remodelling proteins during decidualization of human endometrial stromal cells. J Proteome Res 2010-11-05 [PMID: 20849069] (WB, Human)
• Furmanski O, Gajavelli S, Lee JW et al. Combined extrinsic and intrinsic manipulations exert complementary neuronal enrichment in embryonic rat neural precursor cultures: an in vitro and in vivo analysis. J Comp Neurol;515(1):56-71. 2009-07-01 [PMID: 19399893] (WB, Rat)
• Kim SY, Jun HS, Mead PA et al. Neutrophil stress apoptosis underlie myeloid dysfunction in glycogen storage disease type Ib. Blood;111(12):5704-5711. 2008-01-01 [PMID: 18420828]
• Lahav J et al. Sustained integrin ligation involves extracellular free sulfhydryls enzymatically catalyzed disulfide exchange. Blood 100:2472-8. 2002-01-01 [PMID: 12239158]
• Bennett TA et al. Sulfhydryl regulation of L-selectin shedding: phenylarsine oxide promotes activation-independent L-selectin shedding from leukocytes. J Immunol 164:4120-9. 2000-01-01 [PMID: 10754306]
• Tanaka S et al. Up-regulation of protein-disulfide isomerase in response to hypoxia/brain ischemia its protective effect against apoptotic cell death. J Biol Chem 275:10388-93. 2000-01-01 [PMID: 10744727]
• Courageot J et al. Intracellular degradation of the HIV-1 envelope glycoprotein. Evidence for, some characteristics of, an endoplasmic reticulum degradation pathway. Eur J Biochem 260:482-9. 1999-01-01 [PMID: 10095785]
• Ryser HJ, Levy EM, Mandel R et al. Inhibition of human immunodeficiency virus infection by agents that interfere with thiol-disulfide interchange upon virus-receptor interaction. Proc Natl Acad Sci U S A. 1994-05-01 [PMID: 8183947]
• Kaetzel CS et al. Protein disulphide-isomerase from human placenta rat liver. Purification immunological characterization with monoclonal antibodies. Biochem J 241:39-47. 1987-01-01 [PMID: 3566712]

Reviews for Protein Disulfide Isomerase/P4HB Antibody (NB300-517) (3)

Average Rating: 5

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Other.

reviewed by: Jaehyung (Gus) Cho WB Human 06/20/2011

Summary

• Application: Western Blot
• Sample Tested: HL60 cells transduced with a lentiviral control or PDI shRNA vector., Sample Amount: Lysates of 50,000 HL60 cells
• Species: Human
• Lot: LB138171 and MA151519

reviewed by: Verified Customer WB Human 06/23/2010

Summary

• Application: Western Blot
• Sample Tested: Cell lysate
• Species: Human

reviewed by: Verified Customer WB Other 03/11/2010

Summary

• Application: Western Blot
• Sample Tested: plasma, Sample Amount: 20ul
• Species: Other
• Lot: KE132987

FAQs for Protein Disulfide Isomerase/P4HB Antibody (NB300-517). (Showing 1 - 1 of 1 FAQs).

1. We have recently purchased mouse monoclonal anti-PDI antibody RL90 (cat# NB300-517). From the information available, it appears that this was generated with purified rat PDI serving as immunogen. We would like to know whether there is any information available on the epitope in the PDI molecule recognized by the RL90 antibody? I could not find this information in the description of the antibody.
   • Yes, you are correct: purified rat PDI protein was used as the immunogen for NB300-517, PDI Antibody (RL90). We do not normally epitope map our antibodies, so I think it is unlikely that we know where on the PDI protein this antibody binds. The lab have now confirmed to me that no epitope mapping been carried out for NB300-517, and hence we do not have any additional detail on the binding region of this antibody.

Bioinformatics

• Gene Symbol: P4HB