Human Protein Disulfide Isomerase/P4HB Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human P4HB in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant human P4HB
    Asp18-Lys505
    Accession # P07237
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunoprecipitation
    25 µg/mL
    Conditioned cell culture medium spiked with Recombinant Human Protein Disulfide Isomerase/P4HB (Catalog # 4236-DI), see our available Western blot detection antibodies
  • Immunocytochemistry
    1-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Protein Disulfide Isomerase/P4HB by Western Blot. Western blot shows lysates of PANC‑1 human pancreatic carcinoma cell line and MCF‑7 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Protein Disulfide Isomerase/
P4HB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4236) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Protein Disulfide Isomerase/P4HB at approximately 57 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Protein Disulfide Isomerase/P4HB in HeLa Human Cell Line. Protein Disulfide Isomerase/P4HB was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Protein Disulfide Isomerase/P4HB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4236) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Human Protein Disulfide Isomerase/P4HB by Simple WesternTM. Simple Western lane view shows lysates of PANC‑1 human pancreatic carcinoma cell line and MCF‑7 human breast cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Protein Disulfide Isomerase/P4HB at approximately 59 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Protein Disulfide Isomerase/P4HB Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4236) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: Protein Disulfide Isomerase/P4HB

P4HB (Prolyl 4-hydroxylase beta chain; also PDI) is a 60 kDa member of the protein disulfide isomerase family. As an intracellular homodimer, it forms a tetrameric complex with P4H alpha chains to form an active prolyl 4 hydroxylase. This catalyses the hydroxylation of proline in collagen. On the cell surface, it reduces disulfide bonds in HIV that allow the virus to fuse with CXCR4 and enter susceptible cells. Mature human P4HB is 491 amino acids (aa) in length. It contains two TRX domains (aa 25 134 and 368 475) plus an ER retention sequence (aa 505 508). There is one potential isoform that shows an 11 aa substitution for the first 162 amino acids. Over aa 18 505, human P4HB shares 94% aa identity with mouse P4HB.

  • Long Name:
    Proline 4-Hydroxylase, Beta Polypeptide
  • Entrez Gene IDs:
    5034 (Human); 18453 (Mouse); 25506 (Rat)
  • Alternate Names:
    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
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