Human Calreticulin Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Calreticulin in direct ELISAs and Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Calreticulin
    Met1-Asn180
    Accession # P27797
  • 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
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunocytochemistry
    5-15 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 Calreticulin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and human adrenal gland tissue. PVDF Membrane was probed with 1 µg/mL of Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for Calreticulin at approximately 55 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Immunocytochemistry
Calreticulin in HeLa Human Cell Line. Calreticulin was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Goat Anti-Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) at 10 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 endoplasmic reticula. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry
Calreticulin in Human Pancreas. Calreticulin was detected in immersion fixed paraffin-embedded sections of human pancreas using Goat Anti-Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) at 3 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-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to islet cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human Calreticulin by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and human adrenal gland tissue, loaded at 0.2 mg/mL. A specific band was detected for Calreticulin at approximately 63 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) 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
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • 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: Calreticulin

Human Calreticulin is a 55-60 kDa, 400 amino acid (aa), variably glycosylated intra- and extracellular Ca++-binding lectin that is ubiquitously expressed. It consists of three domains: a 180 aa N-terminal globular region, a 111 aa P-, or proline rich domain, and a 109 aa C-terminus.The 180 aa N-terminus (aa 18-197) is termed Vasostatin. It is unclear if it is ever generated naturally via proteolytic processing. Vasostatin domain has many functions. It binds to RNA (aa 18-27), has autocatalytic phosphorylase activity (aa 77-197), binds to a KxFFKR motif on steroid hormone receptors, and serves as a lectin-type chaperone for ER localized molecules. It also shows anti-angiogenic activity, presumably by binding to laminin carbohydrates and blocking endothelial cell adhesion and proliferation. Human Calreticulin is 94% aa identical to mouse and rat Calreticulin.

  • Entrez Gene IDs:
    811 (Human); 12317 (Mouse); 64202 (Rat)
  • Alternate Names:
    Autoantigen Ro; CALR; Calregulin; Calreticulin; cC1qR; CRP55; CRT; CRTC; Endoplasmic reticulum resident protein 60; ERp60; FLJ26680; grp60; RO; Sicca syndrome antigen A (autoantigen Ro; calreticulin); SSA; SSAHACBP; Vasostatin
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