Calreticulin Antibody (681233) [Unconjugated]

HeLa human cervical epithelial carcinoma cell line was stained with Mouse Anti-Human Calreticulin Monoclonal Antibody (Catalog # MAB38981, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). 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.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Calreticulin knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human/Mouse/Rat Calreticulin Monoclonal Antibody (Catalog # MAB38981) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Calreticulin at approximately 55 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # MAB5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, K562 human chronic myelogenous leukemia cell line, C2C12 mouse myoblast cell line, and NRK rat normal kidney cell line. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Calreticulin Monoclonal Antibody (Catalog # MAB38981) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 formalin fixed paraffin-embedded sections of human prostate using Mouse Anti-Human Calreticulin Monoclonal Antibody (Catalog # MAB38981) 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 plasma membrane and cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu, Mu, Rt
• Applications: WB, IHC, Flow, KO
• Clone: 681233
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Calreticulin Antibody (681233) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Calreticulin
  Glu18-Leu417
  Accession # P27797
• Specificity: Detects human Calreticulin in ELISA. Detects human, mouse and rat Calreticulin in Western blots.
• Source: N/A
• Isotype: IgG2b
• Clonality: Monoclonal
• Host: Mouse
• Gene: CALR

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Immunohistochemistry 8-25 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Knockout Validated
• Control:

  Calreticulin Knockout HeLa Cell Lysate

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 as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 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 Calreticulin Antibody (681233) [Unconjugated]

• 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

Background

Human Calreticulin is a 55‑60 kDa, 400 amino acid, 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 antiangiogenic 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)