Human 2B4/CD244/SLAMF4 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1039

R&D Systems, part of Bio-Techne
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AF1039
AF1039-SP
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Biotin
BAF1039
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Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Agonist Activity, CyTOF-ready, ELISA Capture (Matched Antibody Pair), Flow Cytometry, Immunohistochemistry, Western Blot

Cited:

Immunocytochemistry, Immunohistochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

View all 2B4/CD244/SLAMF4 Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human 2B4/CD244/SLAMF4
Cys22-Arg221
Accession # NP_057466

Specificity

Detects human 2B4/CD244/SLAMF4 in ELISAs and Western blots. In sandwich immunoassays, less than 0.2% cross-reactivity with recombinant mouse 2B4, recombinant human (rh) SLAM, rhCD48, and rhCD84 is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.01 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human 2B4/CD244/SLAMF4 Antibody

Detection of Human 2B4/CD244/SLAMF4 antibody by Western Blot.

Detection of Human 2B4/CD244/SLAMF4 by Western Blot.

Western blot shows lysates of NK-RL human natural killer lymphocytic leukemia cell line, NK-92 human natural killer lymphoma cell line, and Daudi human Burkitt's lymphoma cell line (negative control). PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human 2B4/CD244/SLAMF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1039) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for 2B4/CD244/SLAMF4 at approximately 65 kDa (as indicated). GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of 2B4/CD244/SLAMF4 antibody in Human PBMCs antibody by Flow Cytometry.

Detection of 2B4/CD244/SLAMF4 in Human PBMCs by Flow Cytometry.

Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human NCAM-1/CD56 APC-conjugated Monoclonal Antibody (Catalog # FAB2408A) and either (A) Goat Anti-Human 2B4/CD244/SLAMF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1039) or (B) Normal Goat IgG Control (Catalog # AB-108-C) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107).
2B4/CD244/SLAMF4 antibody in Human Spleen by Immunohistochemistry (IHC-P).

2B4/CD244/SLAMF4 in Human Spleen.

2B4/CD244/SLAMF4 was detected in immersion fixed paraffin-embedded sections of human spleen using Goat Anti-Human 2B4/CD244/SLAMF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1039) at 1 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC004). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to splenocytes. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Applications for Human 2B4/CD244/SLAMF4 Antibody

Application
Recommended Usage

Agonist Activity

Measured by its ability to induce IFN-gamma secretion by NK-92 human natural killer lymphoma cells.
The ED50 for this effect is typically 4‑12 µg/mL.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: Human peripheral blood mononuclear cells

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human spleen

Western Blot

0.5 µg/mL
Sample: NK‑RL human natural killer lymphocytic leukemia cell line and NK‑92 human natural killer lymphoma cell line

Human 2B4/CD244/SLAMF4 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human 2B4/CD244/SLAMF4 Biotinylated Antibody (Catalog # BAF1039)
  • Standard: Recombinant Human 2B4/CD244/SLAMF4 Fc Chimera Protein, CF (Catalog # 1039-2B)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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Qty

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.

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: 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 delta 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

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

Alternate Names

CD244, NAIL, NKR2B4, Nmrk, SLAMF4

Entrez Gene IDs

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

Gene Symbol

CD244

UniProt

NP_057466

Additional 2B4/CD244/SLAMF4 Products

Product Documents for Human 2B4/CD244/SLAMF4 Antibody

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Product Specific Notices for Human 2B4/CD244/SLAMF4 Antibody

For research use only

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