Name: Human ULBP-1 Antibody
Brand: R&D Systems
SKU: AF1380
Price: 469 USD

Species Reactivity

Human

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

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human ULBP-1
Gly26-Pro215
Accession # Q9BZM6

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human ULBP‑1 Fc Chimera (Catalog # 1380-UL)

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.4-1.2 µg/mL of this antibody will block 50% of the binding of 20 ng/mL of biotinylated Recombinant Human ULBP-1 Fc Chimera to immobilized Recombinant Human NKG2D Fc Chimera (Catalog # 1299-NK) coated at 2 µg/mL (100 µL/well). At 30 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

View Larger

Detection of Human ULBP-1 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

View Larger

Detection of Human ULBP-1 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

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Loading...

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-1

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).

References

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

Long Name

UL16 Binding Protein-1

Entrez Gene IDs

80329 (Human); 77777 (Mouse)

Alternate Names

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

Product Datasheets

Product Datasheet

COA

SDS

Catalog #	Availability	Size / Price	Qty
AF1380
AF1380-SP

Human ULBP-1 Antibody