Human VAP-A Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human VAP‑A in direct ELISAs and Western blots. In direct ELISAs, no cross-reactivity with recombinant human VAP-B or recombinant rat VAP-B is observed.
  • Source
    Monoclonal Mouse IgG2A Clone # 604101
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human VAP-A
    Ala2-Met132
    Accession # Q9P0L0
  • 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
    2 µg/mL
    See below
  • Immunohistochemistry
    8-25 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 VAP-A by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line. PVDF Membrane was probed with 2 µg/mL of Mouse Anti-Human VAP-A Monoclonal Antibody (Catalog # MAB5820) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for VAP-A at approximately 33 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1 with 0.05% Tween 20.

Immunohistochemistry
VAP‑A in Human Brain. VAP‑A was detected in immersion fixed paraffin-embedded sections of human brain using Mouse Anti-Human VAP‑A Monoclonal Antibody (Catalog # MAB5820) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 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: VAP-A
Vesicle-associated membrane protein (VAMP)-associated protein A (VAP-A; also VAMP-A and VAP-33) is a 33 kDa, ubiquitously expressed, type IV transmembrane protein belonging to the VAP family of proteins (1). It is found in plasma and ER membranes as well as in intracellular vesicles as a homodimer and a heterodimer with VAP-B. Human VAP-A is synthesized as a 249 amino acid (aa) precursor that contains a 227 aa cytoplasmic domain and a 21 aa transmembrane region. The cytoplasmic domain contains a mobile sperm protein (MSP) domain (aa 13-131) and a coiled-coil region (aa 169-205). Human VAP-A is 97% aa identical to mouse and rat VAP-A. VAP-A and VAP-B recruit FFAT (two phenylalanines in an acidic tract)-motif-containing proteins to the cytosolic surface of ER membranes through a conserved region within their MSP domain, and they have been implicated in regulation of membrane transport, phospholipid biosynthesis, and the unfolded protein response (2, 3). Their ability to interact with lipid-transfer/binding proteins (LT/BPs) may affect the lipid composition of certain cellular membranes (2, 4). VAPs play a critical role in maintaining the structural and functional properties of the Golgi complex (2). Knockdown of VAP reduces the levels of phosphatidylinositol‑4‑phosphate (PI4P), diacylglycerol (DAG), and sphingomyelin (SM) in Golgi membranes and exports pleiotropic effects in Golgi-mediated transport (2). The effects of VAPs are mediated by their interacting FFAT-motif-containing proteins Nir2, OSBP, and CERT (2). VAPs provide a scaffold for these LT/BPs at the ER-Golgi membrane contact sites, thereby affecting the lipid composition of the Golgi membranes and consequently their structural and functional identities (2). VAP-A associates with and regulates the neurite outgrowth-promoting activity of protrudin, a protein that promotes neurite formation (5).
  • References:
    1. Weir, M.L. et al. (1998) Biochem. J. 333:247.
    2. Peretti, D. et al. (2008) Mol. Biol. Cell 19:3871.
    3. Kaiser, S.E. et al. (2005) Structure 13:1035.
    4. Loewen, C.J. et al. (2003) EMBO J. 22:2025.
    5. Saita, S. et al. (2009) J. Biol. Chem. 284:13766.
    6. Long Name:
      VAMP [Vesicle-associated Membrane Protein]-associated Protein A
    7. Entrez Gene IDs:
      9218 (Human); 30960 (Mouse); 58857 (Rat)
    8. Alternate Names:
      hVAP-33; MGC3745,33 kDa VAMP-associated protein; VAMP (vesicle-associated membrane protein)-associated protein A, 33kDa; VAMP-A; VAP-33; VAP33VAMP (vesicle-associated membrane protein)-associated protein A (33kD); VAPA; VAP-A; VAP-AVAMP-associated protein A; vesicle-associated membrane protein-associated protein A
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