Mouse H60 Antibody

Catalog # Availability Size / Price Qty
AF1155
AF1155-SP
Detection of H60 in RAW 264.7 Mouse Cell Line by Flow Cytometry.
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Mouse H60 Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse H60 in direct ELISAs and Western blots.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant mouse H60
Asp30-Gln212
Accession # Q3TDZ7
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Mouse H60 Fc Chimera (Catalog # 1155-H6)
Flow Cytometry
0.25 µg/106 cells
See below
Blockade of Receptor-ligand Interaction
In a functional ELISA, 0.5-2 µg/mL of this antibody will block 50% of the binding of 125 ng/mL of biotinylated Recombinant Mouse NKG2D Fc Chimera to immobilized Recombinant Mouse H60 Fc Chimera (Catalog # 1155-H6) coated at 100 ng/mL (100 µL/well). At 40 μg/mL, this antibody will block >90% of the binding.
 

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.

Scientific Data

Flow Cytometry Detection of H60 antibody in RAW 264.7 Mouse Cell Line antibody by Flow Cytometry. View Larger

Detection of H60 in RAW 264.7 Mouse Cell Line by Flow Cytometry. RAW 264.7 mouse monocyte/macrophage cell line was stained with Goat Anti-Mouse H60 Monoclonal Antibody (Catalog # AF1155, filled histogram) or Goat IgG control antibody (Catalog # AB-108-C, open histogram) followed by anti-Goat IgG PE-conjugated Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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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: H60

H60 was originally described as an immunodominant histocompatibility antigen that is expressed in BALB mice but not in B6 mice. More recently it was reported to function as a ligand for mouse NKG2D, an activating receptor found on NK cells, on some T cell subsets, and on stimulated macrophages. H60 shares approximately 25 percent amino acid identity with the Rae-1 family, a small group of proteins that also function as ligands for mouse NKG2D. H60 and the Rae-1 proteins are distantly related to MHC class I proteins, but they possesses only the alpha 1 and alpha 2 Ig-like domains, and have no capacity to bind peptide or interact with beta 2‑microglobulin. The genes encoding these proteins are not found within the Major Histocompatibility Complex on mouse chromosome 17, but rather map to mouse chromosome 10. Unlike the GPI-linked Rae-1 proteins, H60 appears to be anchored to the membrane via a hydrophobic transmembrane segment. H60 transcripts were found in embryonic tissue, in spleen, and in some transformed cell lines. Transcripts were also observed in mouse skin cells after exposure to carcinogens. Binding of H60 to NKG2D results in the activation of cytolytic activity and/or cytokine production by the NKG2D-expressing effector cells. Ectopic expression of H60 on mouse tumor cell lines resulted in the in vivo rejection of the tumors (1‑6).

References
  1. Malarkannan, S. et al. (1998) J. Immunol. 161:3501.
  2. Diefenbach, A. et al. (2000) Nature Immunol. 1:119.
  3. Cerwenka, A. et al. (2000) Immunity 12:721.
  4. Cerwenka, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:11521.
  5. Diefenbach, A. et al. (2001) Nature 413:165.
  6. NKG2D and its Ligands, www.RnDSystems.com.
Long Name
Histocompatibility 60a
Entrez Gene IDs
15101 (Mouse)
Alternate Names
H60; H60a

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