Human ULBP-4/RAET1E Antibody

Catalog # Availability Size / Price Qty
MAB6285
MAB6285-SP

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Detection of Human ULBP‑4/RAET1E by Western Blot.
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Human ULBP-4/RAET1E Antibody Summary

Species Reactivity
Human
Specificity
Detects human ULBP‑4/RAET1E in direct ELISAs and Western blots. In Western blots, approximately 20% cross‑reactivity with recombinant human (rh) ULBP-2 is observed and no cross-reactivity with rhULBP‑1 or rhULBP-3 is observed.
Source
Monoclonal Mouse IgG2B Clone # 709116
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human ULBP‑4/RAET1E
Gly30-Asp225
Accession # Q8TD07
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
1 µg/mL
See below
Flow Cytometry
0.25 µg/106 cells
HepG2 human hepatocellular carcinoma cell line, U‑87 MG human glioblastoma/astrocytoma cell line, and  HEK293 human embryonic kidney cell line
Immunohistochemistry
8-25 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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

Western Blot Detection of Human ULBP-4/RAET1E antibody by Western Blot. View Larger

Detection of Human ULBP‑4/RAET1E by Western Blot. Western blot shows lysates of HT-29 human colon adenocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human ULBP-4/ RAET1E Monoclonal Antibody (Catalog # MAB6285) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF007). A specific band was detected for ULBP-4/RAET1E at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Flow Cytometry Detection of ULBP‑4/RAET1E antibody in HepG2 Human Cell Line antibody by Flow Cytometry. View Larger

Detection of ULBP‑4/RAET1E in HepG2 Human Cell Line by Flow Cytometry. HepG2 human hepatocellular carcinoma cell line was stained with Mouse Anti-Human ULBP-4/RAET1E Monoclonal Antibody (Catalog # MAB6285, filled histogram) or isotype control antibody (MAB0041, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (F0101B).

Immunohistochemistry ULBP-4/RAET1E antibody in Human Ovarian Cancer Tissue by Immunohistochemistry (IHC-P). View Larger

ULBP‑4/RAET1E in Human Ovarian Cancer Tissue. ULBP-4/RAET1E was detected in immersion fixed paraffin-embedded sections of human ovarian cancer tissue using Mouse Anti-Human ULBP-4/ RAET1E Monoclonal Antibody (Catalog # MAB6285) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; CTS002) and counter-stained with hematoxylin (blue). Specific staining was localized to plasma membranes of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Flow Cytometry View Larger

Detection of ULBP‑4/RAET1E in U87-MG cells by Flow Cytometry. U87-MG cells were stained with Mouse Anti-Human ULBP‑4/RAET1E Monoclonal Antibody (Catalog # MAB6285, filled histogram) or isotype control antibody (Catalog # MAB0041, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Flow Cytometry View Larger

Detection of ULBP‑4/RAET1E in HEK293 cells transfected with Human ULBP-4 (A) vs irrelevant (B) by Flow Cytometry HEK293 cells transfected with Human ULBP-4 (A) vs irrelevant (B) were stained with Mouse Anti-Human ULBP‑4/RAET1E Monoclonal Antibody (Catalog # MAB6285) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.5 mg/mL.
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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: ULBP-4/RAET1E

ULBP-4 (cytomegalovirus glycoprotein UL16 binding protein 4), also called RAET1E (retinoic acid early transcript 1E), Letal (lymphocyte effector cell toxicity activating ligand) and NKG2DL4 (NKG2D ligand 4), is a 40-50 kDa member of the ULBP/RAET1 family of cell surface proteins that function as ligands for NKG2D (1‑6). While most family members are GPI-anchored, only ULBP-4/RAET1E and ULBP-5/RAET1G express a transmembrane form (1, 4, 6, 7). Human ULBP-4 mRNA encodes 263 amino acids (aa) including a 30 aa signal sequence, a 195 aa extracellular domain (ECD), a 23 aa transmembrane domain, and a 15 aa cytoplasmic sequence. A soluble 35 kDa form diverges at aa 208 and is thought to antagonize the transmembrane form (5). Other potential splice variants of 220, 227 and 280 aa are transmembrane proteins (8). Within the ECD, ULBP-4 shares 34-41% aa sequence identity with family members (1, 7). Rodent NKG2D ligands Rae-1 alpha -epsilon are, like human ULBP and MIB proteins, distantly related to MHC class I proteins, but none of the families share significant sequence identity (2, 4). Low expression of ULBP‑4 mRNA is found in normal tissues, with high expression variably reported in the small intestine (3) and skin (4). Expression is stimulated by TNF-alpha and down‑regulated by retinoic acid (3). ULBP-4 is abnormally expressed on most colon cancer and some other tumor cell lines and virus-infected peripheral blood cells (3, 6). ULBP-4 binds and costimulates NKG2D-expressing effector cells including NK cells, NKT cells, gamma δ T cells, and CD8+ alpha beta T cells, activating cytolytic activity and/or cytokine production (3, 4, 7). In some gamma δ T cells, direct ULBP-4 binding to both TCR gamma δ and NKG2D has been demonstrated (6). ULBP-4 is also thought to function as a minor histocompatibility antigen in humans (1).

References
  1. Radosavljevic, M. et al. (2002) Genomics 79:114.
  2. Kondo, M. et al. (2010) Immunogenetics 62:441.
  3. Conejo-Garcia, J.R. et al. (2003) Cancer Biol. Ther. 2:446.
  4. Chalupny, N.J. et al. (2003) Biochem. Biophys. Res. Commun. 305:129.
  5. Cao, W. et al. (2007) J. Biol. Chem. 282:18922.
  6. Kong, Y. et al. (2009) Blood 114:310.
  7. Bacon, L. et al. (2004) J. Immunol. 173:1078.
  8. Cao, W. et al. (2008) Int. Immunol. 20:981.
Long Name
UL16 Binding Protein-4
Entrez Gene IDs
135250 (Human); 379043 (Mouse); 292461 (Rat)
Alternate Names
bA350J20.7; LETAL; LETALRAE-1-like transcript 4; Lymphocyte effector toxicity activation ligand; MGC125309; N2DL4; N2DL-4; NKG2D ligand 4; NKG2DL4; RAET1E; RAET1E2; retinoic acid early transcript 1ERL-4; RL-4; ULBP4; ULBP-4; ULBP4MGC125308

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Citations for Human ULBP-4/RAET1E 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.

5 Citations: Showing 1 - 5
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  1. The NKG2D ligand ULBP4 is not expressed by human monocytes
    Authors: M Lazarova, Y Kim, A Steinle
    PLoS ONE, 2021-02-08;16(2):e0246726.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  2. Clinicopathological relevance of tumor expression of NK group 2 member D ligands in resected non-small cell lung cancer
    Authors: R Okita, A Maeda, K Shimizu, Y Nojima, S Saisho, M Nakata
    Oncotarget, 2019-11-26;10(63):6805-6815.
    Species: Human
    Sample Types: Whole Tissue
    Applications: IHC-P
  3. Targeting chemotherapy-resistant leukemia by combining DNT cellular therapy with conventional chemotherapy
    Authors: B Chen, JB Lee, H Kang, MD Minden, L Zhang
    J. Exp. Clin. Cancer Res., 2018-04-24;37(1):88.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. MicroRNA-519a-3p mediates apoptosis resistance in breast cancer cells and their escape from recognition by natural killer cells
    Authors: C Breunig, J Pahl, M Küblbeck, M Miller, D Antonelli, N Erdem, C Wirth, R Will, A Bott, A Cerwenka, S Wiemann
    Cell Death Dis, 2017-08-03;8(8):e2973.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  5. Adoptively transferred V?9V?2 T cells show potent antitumor effects in a preclinical B cell lymphomagenesis model
    Authors: NA Zumwalde, A Sharma, X Xu, S Ma, CL Schneider, JC Romero-Mas, AW Hudson, A Gendron-Fi, SC Kenney, JE Gumperz
    JCI Insight, 2017-07-06;2(13):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry

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