Calreticulin Antibody [Unconjugated]

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human Calreticulin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3898) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). 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 1.

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 Northern-Lights™ 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.

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.

HeLa human cervical epithelial carcinoma cell line was stained with Goat Anti-Human Calreticulin Affinity-Purified Polyclonal Antibody (Catalog # AF3898, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

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.

Product Discontinued

Calreticulin Antibody [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Calreticulin
  Met1-Asn180
  Accession # P27797
• Specificity: Detects human Calreticulin in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CALR
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Simple Western 10 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

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: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

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

Alternate Names for Calreticulin Antibody [Unconjugated]

• Autoantigen Ro
• CALR
• Calregulin
• Calreticulin
• cC1qR
• CRP55
• CRT
• Endoplasmic reticulum resident protein 60
• epididymis secretory sperm binding protein Li 99n
• ERp60
• FLJ26680
• grp60
• HACBP
• HEL-S-99n
• RO
• Sicca syndrome antigen A (autoantigen Ro; calreticulin)
• SSA
• Vasostatin

Background

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.

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.

Bioinformatics

• Gene Symbol: CALR
• Uniprot:
  • P27797(Human)