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Key Product Details

Species Reactivity

Human

Applications

Blockade of Receptor-ligand Interaction, CyTOF-ready, Flow Cytometry

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2A Clone # 165903

Product Summary for Human ULBP-2/5/6 Antibody

Immunogen

BaF3 mouse pro-B cell line transfected with human ULBP-2
Accession # Q9BZM5

Specificity

Detects human ULBP-2 and human RAET1L/ULBP-6 in direct ELISA and stains cells transfected with human ULBP-2, human ULBP-5 or human RAET1L/ULBP-6 in flow cytometry. In direct ELISA, 11% cross‑reactivity with recombinant human ULBP5 is observed. It does not stain cells transfected with human ULBP-1 or human ULBP-3 in flow cytometry.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2A

Scientific Data Images for Human ULBP-2/5/6 Antibody

Detection of ULBP-2/5/6 antibody in HT1080 Human Cell Line antibody by Flow Cytometry.

Detection of ULBP-2/5/6 in HT1080 Human Cell Line by Flow Cytometry.

HT1080 human fibrosarcoma cell line was stained with Mouse Anti-Human ULBP-2/5/6 Monoclonal Antibody (Catalog # MAB1298, filled histogram) or isotype control antibody (Catalog # MAB0031, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B).
Detection of Human ULBP-2/5/6 by Flow Cytometry

Detection of Human ULBP-2/5/6 by Flow Cytometry

Multiple receptors and ligands are involved in NK cell-mediated lysis of activated CD4+ T cells.Role of (A) activating and (B) inhibitory NK receptors in NK cell degranulation. Left column: representative histograms (of n≥3) for surface expression of ligands on activated (thick black line) and resting CD4+ T cells (thin black line). Isotype-matched control Ig are represented by dashed line (activated CD4+ T) and filled histogram (resting CD4+ T). Middle- and right column: NK and CD4+ T cells were activated for 4 days in vitro as described, and co-cultured for 4 hours with 10 ug/mL mAb (or relevant isotype-matched control Ig). Degranulation is shown for CD56dim (middle column) and CD56bright (right column) NK cells. Representative histograms of surface expression of receptors on activated (thick black line) and resting NK cells (thin black line). Isotype-matched control Ig are represented by dashed line (activated NK) and filled histogram (resting NK). * P<0.05, ** P<0.005, *** P<0.001. (C) Sorted IL-2-activated CD56dim and CD56bright NK cells were co-cultured with 51Cr-labeled activated CD4+ T cells in a 51Cr-release assay with human IgG4 isotype control (•) or anti-NKG2A mAb (○). Data represents n = 3 experiments. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22384114), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human ULBP-2/5/6 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.03 - 0.1 μg/mL of this antibody will block 50% of the binding of 25 ng/mL of biotinylated rhULBP-2/Fc to immobilized rhNKG2D/Fc chimera coated at 2 μg/mL (100 μL/well). At 1 μg/mL, this antibody will achieve 85% blocking in the assay. This antibody will block >70% of the binding of either Recombinant Human ULBP-5 Fc Chimera or Recombinant ULBP-6 Fc Chimera to immobilized Recombinant Human NKG2D Fc Chimera.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HT1080 human fibrosarcoma cell line
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using MAB1298 in the following applications:

Published Applications

Read 66 publications using MAB1298 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Reconstitution Buffer Available:
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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.

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: ULBP-2/5/6

ULBPs activate multiple signaling pathways in primary NK cells, resulting in the production of cytokines and chemokines. Binding of ULBPs ligands to NKG2D induces calcium mobilization and activation of the JAK2, STAT5, ERK and PI3K kinase/Akt signal transduction pathway. The name ULBP derives from the original identification of three proteins, ULBP-1, -2, and -3, as ligands for the human cytomegalovirus glycoprotein UL16; they were designated UL16 binding proteins (ULBP). The genes for ULBPs reside in a cluster of ten related genes, six of which encode potentially functional glycoproteins. ULBP-2 has also been described under the names RaeT1H (retinoic acid early transcript), NKG2DL2, and ALCAN-alpha. ULBP-5 also known as RaeT1G and ULBP-6 also known as RaeT1L. These proteins are distantly related to MHC class I proteins, but they possess only the alpha1 and alpha2 Ig-like domains, and they have no capacity to bind peptide or interact with beta2‑microglobulin. Some family members, including ULBP-2, are anchored to the membrane via a GPI-linkage, whereas others have transmembrane domains. Engagement of NKG2D results in the activation of cytolytic activity and/or cytokine production by these effector cells. The ULBPs are expressed on some tumor cells and have been implicated in tumor surveillance. Over aa 26-217, ULBP-2 shares 92% and 95% aa sequence identity with the human ULBP-5 and ULBP-6, respectively.

References

  1. Cosman, D. et al. (2001) Immunity 14:123.
  2. Kubin, M. et al. (2001) Eur. J. Immunol. 31:1428.
  3. Sutherland, C. et al. (2002) J. Immunol. 168:671.
  4. Steinle, A. et al. (2001) Immunogenetics 53:279.
  5. Sutherland, C. et al. (2001) Immunol. Rev. 181:185.
  6. Pende, D. et al. (2002) Cancer Res. 62:6178.
  7. Radosavljevic, M. et al. (2002) Genomics 79:114.
  8. NKG2D and its Ligands (2002) www.RnDSystems.com.

Long Name

UL16 Binding Protein-2/5/6

Alternate Names

ALCAN-alpha, N2DL2, NKG2DL2, RAET1H, RAET1L, UL16 binding protein 2, ULBP2

UniProt

Product Documents for Human ULBP-2/5/6 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human ULBP-2/5/6 Antibody

For research use only

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