Human 2B4/CD244/SLAMF4 Antibody

Catalog #: MAB10393 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB10393-100
MAB10393-SP

Detection of 2B4/CD244/SLAMF4 in HEK293 Human Cell Line Transfected with Human 2B4/CD244/SLAMF4 and eGFP by Flow Cytometry.

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Human 2B4/CD244/SLAMF4 Antibody Summary

Species Reactivity

Human

Specificity

Detects human 2B4/CD244/SLAMF4 in direct ELISAs.

Source

Monoclonal Mouse IgG_2B Clone # 999602

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

NS0 mouse myeloma cell line transfected with human 2B4/CD244/SLAMF4
Gly19-His222
Accession # NP_057466

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

Flow Cytometry

0.25 µg/10⁶ cells

See below

Blockade of Receptor-ligand Interaction

In a functional ELISA, 100-500 ng/mL of this antibody will block 50% of the binding of 400 ng/mL of Recombinant Human CD48/SLAMF2 (Catalog # 3644-CD) to immobilized Recombinant Human 2B4/CD244/SLAMF4 Fc Chimera (Catalog # 1039-2B) coated at 5 ug/mL (100 µL/well). At 10 ug/mL, this antibody will block >90% of the binding.

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

Flow Cytometry

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Detection of 2B4/CD244/SLAMF4 in HEK293 Human Cell Line Transfected with Human 2B4/CD244/SLAMF4 and eGFP by Flow Cytometry. HEK293 human embryonic kidney cell line transfected with (A) Human 2B4/CD244/SLAMF4 and eGFP or (B) irrelevant protein, was stained with Mouse Anti-Human 2B4/CD244/SLAMF4 Monoclonal Antibody (Catalog # MAB10393) followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). Quadrant markers were set based on Mouse IgG2B isotype control antibody staining (Catalog # MAB0041). View our protocol for Staining Membrane-associated Proteins.

Flow Cytometry

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Detection of 2B4/CD244/SLAMF4 in HEK293 Human Peripheral Blood Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with (A) Mouse Anti-Human 2B4/CD244/SLAMF4 Monoclonal Antibody (Catalog # MAB10393) or (B) Mouse IgG2B isotype control antibody (Catalog # MAB0041) followed by PE-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B) and Mouse anti-Human CD56 APC-conjugated Monoclonal Antibody (Catalog # FAB2408A). View our protocol for Staining Membrane-associated Proteins.

Blockade of Receptor-ligand Interaction

View Larger

CD48/SLAMF2 Binding to 2B4/CD244/SLAMF4 Blocked by Human 2B4/CD244/SLAMF4 Antibody. In a functional ELISA, 100-500 ng/mL of this antibody will block 50% of the binding of 400 ng/mL of Recombinant Human CD48/SLAMF2 (Catalog # 3644-CD) to immobilized Recombinant Human 2B4/CD244/SLAMF4 Fc Chimera (Catalog # 1039-2B) coated at 5 ug/mL (100 µL/well). At 10 ug/mL, this antibody will block >90% of the binding.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile 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: 2B4/CD244/SLAMF4

2B4, also known as CD244 and SLAMF4, is a 66 kDa type I transmembrane glycoprotein in the SLAM subgroup of the CD2 protein family. SLAM family proteins have an extracellular domain (ECD) with two or four Ig-like domains and at least two cytoplasmic immunoreceptor tyrosine-based switch motifs (ITSMs). 2B4 interacts with CD48, while other SLAM family proteins interact homophilically (1‑4). Mature human 2B4 consists of a 208 amino acid (aa) ECD with two Ig-like domains, a 21 aa transmembrane segment, and a 120 aa cytoplasmic domain with four ITSMs (5, 6). Three additional splice variants of human 2B4 have deletions of the short region between the Ig-like domains, the second Ig-like domain, or a portion of the cytoplasmic tail. Within the ECD, human 2B4 shares 46% and 40% aa sequence identity with mouse and rat 2B4, respectively. The ECD of human 2B4 shares 17%‑24% aa sequence identity with comparable regions of human CD2 family members BLAME, CD2F-10, CD84, CD229, CRACC, NTB-A, and SLAM. 2B4 is expressed on all NK cells, gamma δ T cells, monocytes, some CD4⁺ and CD8⁺ T cells, and some dendritic cells (7). CD48 mediates 2B4⁺ cell interactions with nearly all hematopoietic cell types, including cells of the same type (8‑10). 2B4/CD48 signaling cooperates with other receptor systems to either promote or inhibit NK and CD8⁺ T cell activation (7‑13). The inhibitory activities are distinct from those of MHC I restricted inhibitory NK cell receptors (12, 13). Ligation of 2B4 with antibodies or CD48 constructs can either directly trigger inhibitory signaling or disrupt an inhibitory interaction, leading to cellular activation (9, 12). The inhibitory effect is associated with the long form of 2B4, while the activation is associated with the short form (9, 14). 2B4 can also induce signaling through CD48 (10, 15).

References

Bhat, R. et al. (2006) J. Leukoc. Biol. 79:417.
Veillette, A. (2006) Nat. Rev. Immunol. 6:56.
McNerney, M.E. et al. (2005) Mol. Immunol. 42:489.
Assarsson, E. et al. (2005) J. Immunol. 175:2045.
Boles, K.S. et al. (1999) Tissue Antigens 54:27.
Kubin, M.Z. et al. (1999) Eur. J. Immunol. 29:3466.
Nakajima, H. et al. (1999) Eur. J. Immunol. 29:1676.
Lee, K.M. et al. (2006) Blood 107:3181.
Mooney, J.M. et al. (2004) J. Immunol. 173:3953.
Assarsson, E. et al. (2004) J. Immunol. 173:174.
Bryceson, Y.T. et al. (2006) Blood 107:159.
Lee, K-M. et al. (2004) J. Exp. Med. 199:1245.
McNerney, M.E. et al. (2005) Blood 106:1337.
Schatzle, J.D. et al. (1999) Proc. Natl. Acad. Sci. USA 96:3870.
Messmer, B. et al. (2006) J. Immunol. 176:4646.

Entrez Gene IDs

51744 (Human); 18106 (Mouse); 685808 (Rat); 102132700 (Cynomolgus Monkey)

Alternate Names

2B4; CD244 antigen; CD244 molecule, natural killer cell receptor 2B4; CD244; h2B4; NAIL; NAILNK cell activation-inducing ligand; natural killer cell receptor 2B4; NKR2B4; NKR2B4NK cell type I receptor protein 2B4,2B4CD244 natural killer cell receptor 2B4; Nmrk; SLAMF4; SLAMF4NK cell activation inducing ligand NAIL