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Human ULBP-3 Antibody

Catalog #: AF1517
11 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
AF1517
AF1517-SP
Detection of Human ULBP-3 by Flow Cytometry
2 Images
Product Details
Citations (11)
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Human ULBP-3 Antibody Summary

Species Reactivity
Human
Specificity
Detects human ULBP-3 in direct ELISAs and Western blots. In these formats, approximately 5% cross‑reactivity with recombinant human (rh) ULBP-2 is observed and less than 1% cross-reactivity with rhULBP-1 is observed.
Source
Polyclonal Goat IgG
Purification
Antigen Affinity-purified
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ULBP-3
Gly27-Pro216
Accession # NP_078794
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.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration
Sample
Western Blot
0.1 µg/mL
Recombinant Human ULBP‑3 Fc Chimera (Catalog # 1517-UL)
Blockade of Receptor-ligand Interaction
In a functional ELISA, 1-3 µg/mL of this antibody will block 50% of the binding of 20 ng/mL of biotinylated Recombinant Human ULBP-3 Fc Chimera to immobilized Recombinant Human NKG2D Fc Chimera (Catalog # 1299-NK) coated at 2 µg/mL (100 µL/well). At 100 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 Human ULBP-3 by Flow Cytometry View Larger

Detection of Human ULBP-3 by Flow Cytometry Rh159 interferes with intracellular transport of NKG2DL.A) Association with Rh159 prevents intracellular transport of MICB. U373-MICB cells were transduced with adenovectors (MOI = 80) expressing either GFP (AdGFP) or FLAG-tagged Rh159 (AdRh159FL) under control of tetracycline-dependent transactivator provided by co-transduced AdtTA (MOI = 20). At 24 hpi cells were metabolically labeled for 30 min with [35S]cysteine + [35S]methionine. Upon chasing the label for the indicated times (h), cells were lysed and MICB was immunoprecipitated with anti–MICB mAb. Precipitates were either digested with EndoH (+) or mock treated (-) followed by SDS-PAGE and autoradiography. (S) indicates EndoH-deglycosylated proteins. B) Rh159 co-immunoprecipitates with MICB. U373-ULBP3 (ULBP3, left panel) or U373-MICB (MICB, right panel) cells were lysed at 48 h post-transduction with AdRh159FL (Rh159) or an adenovector expressing FLAG-tagged SVV ORF 61 (SVV61) used as a negative control. MICB and ULBP3 were immunoprecipitated with anti–MICB and anti-ULBP3 mouse and goat mAbs, respectively, then immunoblotted with mouse anti-FLAG mAb. The mouse IgG heavy chain (55kDa) is indicated (HC). Input lanes were loaded with 10% total lysate used in immunoprecipitation and immunoblotted with mAbs for FLAG and GAPDH. The results shown are representative of two independent experiments. C) Rh159 reduces steady state levels of MICB. U373-MICB cells were lysed at 48 h post-transduction with the indicated Ad-vectors. Lysates were digested with EndoH (+) or mock treated (-) then immunoblotted with mAbs for MICB, FLAG or GAPDH. Note that both MICB and Rh159 are EndoH sensitive consistent with ER localization. The results shown are representative of two independent experiments. D-E) Rh159 reduces surface expression of MICA, MICB, ULBP1 and ULBP2 but not ULBP3. U373-NKG2DL cells were transduced with AdRh159FL or AdGFP as in A) but for 48 h. Cells were then lysed and immunoblotted with mAbs for FLAG and GAPDH (D), or stained with antibodies specific for the indicated proteins, or isotype control (dotted) and analyzed by flow cytometry. The results shown are representative of three or more independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27580123), licensed under a CC-BY license. Not internally tested by R&D Systems.

Flow Cytometry Detection of Human ULBP-3 by Flow Cytometry View Larger

Detection of Human ULBP-3 by Flow Cytometry Rh159 interferes with intracellular transport of NKG2DL.A) Association with Rh159 prevents intracellular transport of MICB. U373-MICB cells were transduced with adenovectors (MOI = 80) expressing either GFP (AdGFP) or FLAG-tagged Rh159 (AdRh159FL) under control of tetracycline-dependent transactivator provided by co-transduced AdtTA (MOI = 20). At 24 hpi cells were metabolically labeled for 30 min with [35S]cysteine + [35S]methionine. Upon chasing the label for the indicated times (h), cells were lysed and MICB was immunoprecipitated with anti–MICB mAb. Precipitates were either digested with EndoH (+) or mock treated (-) followed by SDS-PAGE and autoradiography. (S) indicates EndoH-deglycosylated proteins. B) Rh159 co-immunoprecipitates with MICB. U373-ULBP3 (ULBP3, left panel) or U373-MICB (MICB, right panel) cells were lysed at 48 h post-transduction with AdRh159FL (Rh159) or an adenovector expressing FLAG-tagged SVV ORF 61 (SVV61) used as a negative control. MICB and ULBP3 were immunoprecipitated with anti–MICB and anti-ULBP3 mouse and goat mAbs, respectively, then immunoblotted with mouse anti-FLAG mAb. The mouse IgG heavy chain (55kDa) is indicated (HC). Input lanes were loaded with 10% total lysate used in immunoprecipitation and immunoblotted with mAbs for FLAG and GAPDH. The results shown are representative of two independent experiments. C) Rh159 reduces steady state levels of MICB. U373-MICB cells were lysed at 48 h post-transduction with the indicated Ad-vectors. Lysates were digested with EndoH (+) or mock treated (-) then immunoblotted with mAbs for MICB, FLAG or GAPDH. Note that both MICB and Rh159 are EndoH sensitive consistent with ER localization. The results shown are representative of two independent experiments. D-E) Rh159 reduces surface expression of MICA, MICB, ULBP1 and ULBP2 but not ULBP3. U373-NKG2DL cells were transduced with AdRh159FL or AdGFP as in A) but for 48 h. Cells were then lysed and immunoblotted with mAbs for FLAG and GAPDH (D), or stained with antibodies specific for the indicated proteins, or isotype control (dotted) and analyzed by flow cytometry. The results shown are representative of three or more independent experiments. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/27580123), licensed under a CC-BY license. Not internally tested by R&D Systems.

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
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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-3

ULBP-3 is a member of a family of cell-surface proteins that function as ligands for human NKG2D. ULBP-3 has also been described under the names RaeT1N (retinoic acid early transcript), NKG2DL3, and ALCAN-gamma. The name ULBP-3 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 gene for ULBP-3 resides in a cluster of ten related genes, six of which encode potentially functional glycoproteins. Amino acid sequence identity within this family ranges from 30-60%. These proteins are distantly related to MHC class I proteins, but they possess only the alpha 1 and alpha 2 Ig-like domains, and they have no capacity to bind peptide or interact with beta 2‑microglobulin. Some family members, including ULBP-3, are anchored to the membrane via a GPI-linkage, whereas others have transmembrane domains. ULBP‑3 and several other family members are known to bind to human NKG2D, an activating receptor expressed on NK cells, NKT cells, gamma δ T cells, and CD8+ alpha beta T cells. 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 (1-7).

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) http://www.RnDSystems.com/.
Long Name
UL16 Binding Protein-3
Entrez Gene IDs
79465 (Human)
Alternate Names
NKG2D ligand 3; RAET1N; UL16 binding protein 3; ULBP3; ULBP-3

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Citations for Human ULBP-3 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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  1. Soluble NKG2DLs Are Elevated in Breast Cancer Patients and Associate with Disease Outcome
    Authors: Seller, A;Tegeler, CM;Mauermann, J;Schreiber, T;Hagelstein, I;Liebel, K;Koch, A;Heitmann, JS;Greiner, SM;Hayn, C;Dannehl, D;Engler, T;Hartkopf, AD;Hahn, M;Brucker, SY;Salih, HR;Märklin, M;
    International journal of molecular sciences
    Species: Human
    Sample Types: Serum
    Applications: ELISA Capture
  2. CRISPR activation screen identifies BCL-2 proteins and B3GNT2 as drivers of cancer resistance to T cell-mediated cytotoxicity
    Authors: J Joung, PC Kirchgatte, A Singh, JH Cho, SP Nety, RC Larson, RK Macrae, R Deasy, YY Tseng, MV Maus, F Zhang
    Nature Communications, 2022-03-25;13(1):1606.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Natural Killer Group 2D Ligand Depletion Reconstitutes Natural Killer Cell Immunosurveillance of Head and Neck Squamous Cell Carcinoma
    Authors: S Weil, S Memmer, A Lechner, V Huppert, A Giannattas, T Becker, A Müller-Run, K Lampe, D Beutner, A Quaas, R Schubert, E Herrmann, A Steinle, U Koehl, L Walter, MS von Bergwe, J Koch
    Front Immunol, 2017-04-10;8(0):387.
    Applications: ELISA Development (Capture)
  4. Natural Killer Cell Evasion Is Essential for Infection by Rhesus Cytomegalovirus
    PLoS Pathog, 2016-08-31;12(8):e1005868.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Varicella-Zoster Virus and Herpes Simplex Virus 1 Differentially Modulate NKG2D Ligand Expression during Productive Infection
    Authors: Tessa M. Campbell, Brian P. McSharry, Megan Steain, Barry Slobedman, Allison Abendroth
    Journal of Virology
  6. Cytotoxicity and infiltration of human NK cells in in vivo-like tumor spheroids.
    Authors: Giannattasio A, Weil S, Kloess S, Ansari N, Stelzer E, Cerwenka A, Steinle A, Koehl U, Koch J
    BMC Cancer, 2015-05-03;15(0):351.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: ELISA Development (Capture)
  7. Immunohistochemical validation and expression profiling of NKG2D ligands in a wide spectrum of human epithelial neoplasms.
    Authors: Fujita H, Hatanaka Y, Sutoh Y, Suzuki Y, Oba K, Hatanaka K, Mitsuhashi T, Otsuka N, Fugo K, Kasahara M, Matsuno Y
    J Histochem Cytochem, 2014-12-03;63(3):217-27.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  8. Comprehensive Analysis of NKG2D Ligand Expression and Release in Leukemia: Implications for NKG2D-Mediated NK Cell Responses
    Authors: Julia Hilpert, Ludger Grosse-Hovest, Frank Grünebach, Corina Buechele, Tina Nuebling, Tobias Raum et al.
    The Journal of Immunology
  9. The human NKG2D ligand ULBP2 can be expressed at the cell surface with or without a GPI anchor and both forms can activate NK cells.
    Authors: Fernandez-Messina L, Ashiru O, Aguera-Gonzalez S, Reyburn HT, Vales-Gomez M
    J. Cell. Sci., 2011-01-11;124(0):321-7.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC, Immunoprecipitation
  10. Intracellular Sequestration of the NKG2D Ligand ULBP3 by Human Cytomegalovirus
    Authors: Neil J. Bennett, Omodele Ashiru, Fiona J. E. Morgan, Yin Pang, Georgina Okecha, Rob A. Eagle et al.
    The Journal of Immunology
  11. NKG2D ligand expression in human colorectal cancer reveals associations with prognosis and evidence for immunoediting.
    Authors: McGilvray RW, Eagle RA, Watson NF, Al-Attar A, Ball G, Jafferji I, Trowsdale J, Durrant LG
    Clin. Cancer Res., 2009-10-27;15(22):6993-7002.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P

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