ERp57/PDIA3 Antibody

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HEK293 human embryonic kidney cell line, and Hepa 1-6 mouse hepatoma cell line. PVDF membrane was probed with 0.2 µg/mL of Sheep Anti-Human ERp57/PDIA3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8219) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for ERp57/PDIA3 at approximately 57 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

ERp57/PDIA3 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human ERp57/PDIA3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8219) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

ERp57/PDIA3 was detected in immersion fixed paraffin-embedded sections of human liver using Sheep Anti-Human ERp57/PDIA3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8219) at 1 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm of hepatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Protein kinase A (PKA) catalytic subunit alpha is localized within mitochondria of HeLa cells. (A) Representative immunoblots (n = 3) of PKA catalytic (cat) subunit, PKA regulatory (reg) subunit, the mitochondrial protein SDHa and of the cytosolic protein alpha -Tubulin with corresponding total protein load (TPL) in total cell lysate (TCL), cytosolic (Cyto) and mitochondrial (Mito) fractions isolated from HeLa cells. (B) Representative immunoblots (n = 3) of PKA cat, PKA reg, SDHa and TOM20 with corresponding total protein load (TPL) in mitochondrial fractions isolated from HeLa cells and treated as indicated. (C) Representative micrographs (n = 6) of HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc and labelled with anti-Myc (green) and anti-TOM20 (red) as a mitochondrial marker. Scale bars = 5 μm. (D) Representative immunoblots (n = 3) of Myc, SDHa and alpha -Tubulin with corresponding total protein load (TPL) in TCL, cyto and mito fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc. (E) Representative immunoblots (n = 3) of Myc, TOM20, SDHa and SOD2 with corresponding total protein load (TPL) in mitochondrial enriched fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc and treated as indicated. (F) Representative immunoblots (n = 3) of Myc, PKA Cat, ATP5a, MFN2 and ERp57 with corresponding TPL in pellet and supernatant (SN) obtained after treatment of mitochondria isolated from HeLa cells expressing PKA-Myc or mt-PKA-Myc and treated as indicated. (G) Representative immunoblots (n = 5) of phospho-PKA substrates and Myc with corresponding total protein load (TPL) in mitochondrial enriched fractions isolated from HeLa cells expressing empty vector (pcDNA), PKA-Myc or mt-PKA-Myc. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/33167377), licensed under a CC-BY license. Not internally tested by R&D Systems.

Mitochondrial matrix-localized Src regulates mitochondrial shape. a Representative immunoblotting (n = 3) of Src, cytosolic tubulin and mitochondrial SDHA in total cell lysate (TCL), cytosol- (cyto) and mitochondria- (mito) enriched fractions of Src+/+ and Src−/− MEFs, HEK293 and HeLa. b Representative immunoblotting (n = 3) of Src, the IMM protein SDHA and the OMM protein TOM20 in mitochondria-enriched fractions of HeLa cells treated as indicated. c Representative immunoblotting (n = 4) of Src, Smac-diablo, ATP5a and ERp57 in supernatant (SN) and pellet obtained from mitochondria-enriched fractions of HeLa cells treated with digitonin as indicated. d Representative micrographs (n = 3) of the plasma membrane marker wheat germ agglutinin (WGA), V5 and TOM20 labeling in Src+/+ and Src−/− mouse embryonic fibroblasts (MEFs) expressing pcDNA, Src-V5, mitochondria-targeted mtSrc-V5 and plasma membrane-targeted pmSrc-V5. e Quantitative analysis of mitochondrial morphology in region of interest (ROI) as shown in c. Scale bars: 20 μm. Data are presented as mean ± S.E.M. Data with different letters are statistically different (p < 0.05), according to one-way ANOVA followed by Tukey’s post hoc test Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35637383), licensed under a CC-BY license. Not internally tested by R&D Systems.

Mitochondrial matrix-localized Src regulates mitochondrial shape. a Representative immunoblotting (n = 3) of Src, cytosolic tubulin and mitochondrial SDHA in total cell lysate (TCL), cytosol- (cyto) and mitochondria- (mito) enriched fractions of Src+/+ and Src−/− MEFs, HEK293 and HeLa. b Representative immunoblotting (n = 3) of Src, the IMM protein SDHA and the OMM protein TOM20 in mitochondria-enriched fractions of HeLa cells treated as indicated. c Representative immunoblotting (n = 4) of Src, Smac-diablo, ATP5a and ERp57 in supernatant (SN) and pellet obtained from mitochondria-enriched fractions of HeLa cells treated with digitonin as indicated. d Representative micrographs (n = 3) of the plasma membrane marker wheat germ agglutinin (WGA), V5 and TOM20 labeling in Src+/+ and Src−/− mouse embryonic fibroblasts (MEFs) expressing pcDNA, Src-V5, mitochondria-targeted mtSrc-V5 and plasma membrane-targeted pmSrc-V5. e Quantitative analysis of mitochondrial morphology in region of interest (ROI) as shown in c. Scale bars: 20 μm. Data are presented as mean ± S.E.M. Data with different letters are statistically different (p < 0.05), according to one-way ANOVA followed by Tukey’s post hoc test Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/35637383), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, IHC, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

ERp57/PDIA3 Antibody Summary

• Immunogen: E. coli-derived recombinant ERp57/PDIA3
  Arg107-Lys366
  Accession # P30101
• Specificity: Detects human ERp57/PDIA3 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: PDIA3
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 0.2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ERp57/PDIA3 Antibody

• Disulfide isomerase ER-60
• EC 5.3.4.1
• endoplasmic reticulum P58
• Endoplasmic reticulum resident protein 57
• Endoplasmic reticulum resident protein 60
• ER protein 60
• ER60
• ERp57
• ERp5758 kDa glucose-regulated protein
• ERp60
• ERp6058 kDa microsomal protein
• ERp61
• glucose regulated protein, 58kDa
• GRP57
• GRP58
• GRP58ER protein 57
• HsT17083
• P58
• PDIA3
• phospholipase C-alpha
• PI-PLC
• protein disulfide isomerase family A, member 3
• protein disulfide isomerase-associated 3
• protein disulfide-isomerase A3

Background

ERp57 (Endoplasmic reticulum resident protein 57; also Protein disulfide-isomerase A3/PDIA3) is a 57-60 kDa member of the protein disulfide isomerase family of proteins.  Knockdown of ERp57 or antibody-targeted inhibition of the secreted form significantly impaired the secretion and accumulation of extracellular matrix. ERp57 is highly similar to other PDI family members in amino acid sequence and structural/functional domain organization. This protein plays an important role in endoplasmic reticulum quality control of newly synthesized glycoproteins, is critical in major histocompatability complex class I assembly and regulates gene expression. ERp57 has been implicated in human pathologies including cancer and Alzheimer's disease. Within aa 107 - 366, human ERp57  shares 92% aa sequence identity with mouse ERp57.

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 ERp57/PDIA3 Antibody (AF8219)(2)

Publications using AF8219

• Olivier Lurette, Hala Guedouari, Jordan L. Morris, Rebeca Martín-Jiménez, Julie-Pier Robichaud, Geneviève Hamel-Côté et al. Mitochondrial matrix-localized Src kinase regulates mitochondrial morphology Cellular and Molecular Life Sciences 6/1/2022 [PMID: 35637383]
• S Larsson, U Voss Neuroprotective effects of vitamin D on high fat diet- and palmitic acid-induced enteric neuronal loss in mice BMC Gastroenterol, 2018-11-21;18(1):175. 2018-11-21 [PMID: 30463517] (ICC, Mouse)

Bioinformatics

• Gene Symbol: PDIA3
• Uniprot:
  • P30101()