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ABS1655 Sigma-Aldrich

Anti-PDI Antibody, arginylated (Nt-Asp18)

Detect arginylated mature PDI using this rabbit polyclonal Anti-PDI, arginylated (Nt-Asp18), Cat. No. ABS1655, validated for use in ELISA, Immunocytochemistry, and Western Blotting.

Synonym: Cellular thyroid hormone-binding protein, Nt-Asp18 arginylated, P4Hbeta, Nt-Asp18 arginylated, PDI, Nt-Asp18 arginylated, Prolyl 4-hydroxylase subunit beta, Nt-Asp18 arginylated, Protein disulfide-isomerase, Nt-Asp18 arginylated, p55, Nt-Asp18 arginylated

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Properties

Related Categories Antibodies, Primary Antibodies More...
format   Affinity Purified
molecular weight   ~57 kDa observed. 55.45 kDa calculated. Uncharacterized bands may be observed in some lysate(s).
application(s)   ELISA
  Immunocytochemistry
  Western Blotting
clone   Polyclonal Antibody
purification method   Affinity purified.
epitope   N-terminus
concentration   Please refer to lot specific datasheet.
host   Rabbit
species reactivity   Human
NCBI accession no.   NP_000909
UniProt accession no.   P07237
gene symbol   P4HB(5034)
  CLCRP1
  ERBA2L
  PDI
  PDIA1
  PO4DB
material size   100 μg
shipped in   ambient
storage conditions   Stable for 1 year at 2-8°C from date of receipt.

Description

Background information

Protein disulfide-isomerase (EC 5.3.4.1; UniProt P07237; also known as Cellular thyroid hormone-binding protein, p55, P4Hbeta, PDI, Prolyl 4-hydroxylase subunit beta) is encoded by the P4HB (also known as CLCRP1, ERBA2L, PDI, PDIA1, PO4DB) gene (Gene ID 5034) in human. Protein disulphide isomerase (PDI), BiP (GRP78 or heat shock 70 kDa protein 5), and calreticulin (CRT) are ER lumen folding factors that, together with other PTM enzymes, assist the process of newly synthesized proteins before their release from ER. PDI, BiP, and CRT themselves are subject to posttranslational signal peptide removal after their synthesis, exposing D18, E19, and E18 at the newly formed N-terminus, respectively. The N-terminus D and E residues of the mature proteins are permissive to arginine-tRNA-protein transferase 1/ATE1-catalyzed arginylation, forming arginylated mature proteins (R-DPI, R-BiP, and R-CRT) with altered cellular localization to allow their participation in non-ER functions. Although only the basal levels of R-PDI and R-CRT, but not R-BiP, are generally detectible in non-stimulated cells, upregulated N-terminal arginylation of all three proteins is observed upon cytosolic dsDNA exposure and proteasomal inhibition, indicating a shared role in innate immune responses to invading microbes. In addition, ER stress induction by thapsigargin treatment synergistically boosts proteasomal inhibition-induced upregulation of cellular levels of R-DPI, R-BiP, and R-CRT, suggesting that ER stress accelerates the supply of ER lumenal DPI, BiP, and CRT for N-terminal arginylation.

Physical form

Purified rabbit polyclonal antibody in PBS with 0.05% sodium azide.

Application

Immunocytochemistry Analysis: 1:100 dilution from a representative lot detected PDI Nt-Asp18 arginylation induction in (18-hr 3 µM MG132 and 200 nM thapsigargin) treated HeLa cells (Courtesy of Yong Tae Kwon, Ph.D. , Seoul National University, Korea).

Western Blotting Analysis: 0.2 µg/mL from a representative lot detected PDI Nt-Asp18 arginylation induction in (18-hr 3 µM MG132 and 200 nM thapsigargin) treated HeLa cells (Courtesy of Yong Tae Kwon, Ph.D. , Seoul National University, Korea).

ELISA Analysis: A representative lot detected the immunogen peptide, but not the control peptide without arginylation at the N-terminal Asp18 (Cha-Molstad, H., et al. (2015). Nat. Cell Biol. 17(7):917-929).

Western Blotting Analysis: A representative lot detected PDI Nt-Asp18 arginylation induction upon poly(dA:dT) transfection or arginine-tRNA-protein transferase 1 (ATE1) 1A7A isoform overexpression in HeLa cells. Combined proteasome inhibition and ER stress induction by an 18-hr 10 µM MG132 and 100 nM thapsigargin treatment synergized the two drugs' efficacy toward cellular PDI Nt-Asp18 arginylation induction (Cha-Molstad, H., et al. (2015). Nat. Cell Biol. 17(7):917-929).

Specificity

This rabbit polyclonal antibody specifically detected the immunogen peptide, but not the control peptide without arginylation at the N-terminal Asp18 (Cha-Molstad, H., et al. (2015). Nat. Cell Biol. 17(7):917-929).

Immunogen

Linear peptide corresponding to the N-terminal sequence of arginylated mature human PDI.

Species Reactivity Notes

Human. Predicted to react with Chimpanzee based on 100% sequence homology.

Quality Assurance

Evaluated by Western Blotting in treated HEK293 cells.

Western Blotting Analysis: 1 µg/mL of this antibody detected PDI Nt-Asp18 arginylation induction in 7.5 µg of lysate from (17-hr 3 µM MG132 and 200 nM thapsigargin) treated HEK293 cells.

Usage Statement

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles

Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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