ULBP-1 Antibody [Unconjugated]

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 (//pubmed.ncbi.nlm.nih.gov/27580123), licensed under a CC-BY license. Not internally tested by R&D Systems.

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 (//pubmed.ncbi.nlm.nih.gov/27580123), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, B/N
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

ULBP-1 Antibody [Unconjugated] Summary

• Immunogen: S. frugiperda insect ovarian cell line Sf 21-derived recombinant human ULBP-1
  Gly26-Pro215
  Accession # Q9BZM6
• Specificity: Detects human ULBP-1 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross-reactivity with recombinant human (rh) ULBP-2 and rhULBP-3 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: ULBP1
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Blockade of Receptor-ligand Interaction
  • Western Blot 0.1 ug/mL

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: Reconstitute at 0.2 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 ULBP-1 Antibody [Unconjugated]

• ALCAN-beta
• MULT-1
• N2DL1
• N2DL-1
• NKG2D ligand 1
• NKG2DL
• NKG2DL1
• RAET1I
• RAET1Ialcan-beta
• Retinoic acid early transcript 1I
• UL16 binding protein 1
• UL16-binding protein 1
• ULBP1
• ULBP-1

Background

ULBP-1 is a member of a family of cell-surface proteins that function as ligands for human NKG2D. ULBP-1 has also been described under the names RaeT1I (retinoic acid early transcript), ALCAN-beta, and NKG2DL1. The name ULBP-1 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-1 resides in a cluster of ten related genes, six of which encode potentially functional glycoproteins. Amino acid sequence identity within this family ranges from 30‑95%. 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. They are anchored to the membrane via a GPI-linkage. ULBP-1 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. ULBP-1 is expressed on some tumor cells and has been implicated in tumor surveillance (1‑8).

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.

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.

Publications for ULBP-1 Antibody (AF1380)(5)

Publications using AF1380

• Natural Killer Cell Evasion Is Essential for Infection by Rhesus Cytomegalovirus PLoS Pathog, 2016-08-31;12(8):e1005868. 2016-08-31 [PMID: 27580123] (Flow Cytometry, Human)
• Leung W, Vong Q, Lin W, Bouck D, Wendt S, Sullivan E, Li Y, Bari R, Chen T, Leung W PRL-3 mediates the protein maturation of ULBP2 by regulating the tyrosine phosphorylation of HSP60. J Immunol, 2015-02-16;194(6):2930-41. 2015-02-16 [PMID: 25687758] (Flow Cytometry, Western Blot, Human)
• Fujita H, Hatanaka Y, Sutoh Y, Suzuki Y, Oba K, Hatanaka K, Mitsuhashi T, Otsuka N, Fugo K, Kasahara M, Matsuno Y Immunohistochemical validation and expression profiling of NKG2D ligands in a wide spectrum of human epithelial neoplasms. J Histochem Cytochem, 2014-12-03;63(3):217-27. 2014-12-03 [PMID: 25473094] (IHC-P, Human)
• Christine L. Schneider, Amy W. Hudson The Human Herpesvirus-7 (HHV-7) U21 Immunoevasin Subverts NK-Mediated Cytoxicity through Modulation of MICA and MICB PLoS Pathogens 11/10/2011 [PMID: 22102813]
• McGilvray RW, Eagle RA, Watson NF, Al-Attar A, Ball G, Jafferji I, Trowsdale J, Durrant LG NKG2D ligand expression in human colorectal cancer reveals associations with prognosis and evidence for immunoediting. Clin. Cancer Res., 2009-10-27;15(22):6993-7002. 2009-10-27 [PMID: 19861434] (IHC-P, Human)

Bioinformatics

• Gene Symbol: ULBP1
• Uniprot:
  • Q9BZM6(Human)