2B4/CD244/SLAMF4 Antibody (025) - Azide and BSA Free

Western Blot: 2B4/CD244/SLAMF4 Antibody (025) [NBP2-89232] - Lane A: Daudi Whole Cell Lysate Lysates/proteins at 30 ug per lane.Secondary Goat Anti-Rabbit IgG H&L (Dylight800) at 1/10000 dilution.Developed using the Odyssey technique. Performed under reducing conditions.Predicted band size:42 kDa Observed band size:28 kDa (We are unsure as to the identity of these extra bands.)

Immunoprecipitation: 2B4/CD244/SLAMF4 Antibody (025) [NBP2-89232] - Lane A:0.5 mg Daudi Whole Cell Lysate 2 uL anti-2B4 rabbit monoclonal antibody and 15 ul of 50 % Protein G agarose. Primary antibody: Anti-2B4 rabbit monoclonal antibody,at 1:200 dilution Secondary antibody: Dylight 800-labeled antibody to rabbit IgG (H+L), at 1:5000 dilution Developed using the odssey technique. Performed under reducing conditions. Predicted band size: 42 kDa Observed band size: 28 kDa

• Reactivity: Hu
• Applications: WB, ELISA, IP, ELISA
• Clone: 025
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: Azide and BSA Free

2B4/CD244/SLAMF4 Antibody (025) - Azide and BSA Free Summary

• Description: This antibody can be stored at 2C-8C for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20C to -80C. Avoid repeated freeze-thaw cycles.
• Additional Information: Recombinant Monoclonal Antibody
• Immunogen: This antibody was obtained from a rabbit immunized with purified, recombinant Human 2B4/CD244/SLAMF4 (Accession#: NP_057466.1; Met1-Arg221).
• Source: HEK293
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit
• Gene: CD244
• Purity: Protein A purified

Applications/Dilutions

• Dilutions:
  • ELISA
  • Immunoprecipitation 1-4 uL/mg of lysate
  • Sandwich ELISA Capture
  • Western Blot 1:500-1:2000
• Application Notes: This antibody will detect Human 2B4/CD244/SLAMF4 in ELISA pair set (Catalog: # NBP2-78845).

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: 0.2 um filtered solution in PBS
• Preservative: No Preservative
• Purity: Protein A purified

Alternate Names for 2B4/CD244/SLAMF4 Antibody (025) - Azide and BSA Free

• 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

Background

2B4, also known as CD244 and SLAMF4, is a type I transmembrane glycoprotein that is member of the signaling lymphocyte activation molecule (SLAM) subfamily in immunoglobulin (Ig) superfamily (1,2). The 2B4 protein plays a role in immune system associated diseases including the recognition and clearance of cancer cells, infected cells, and key allergy cells (1). 2B4 is expressed as a heterophilic receptor on the cell membrane of natural killer (NK) cells in addition to on monocytes and basophils, gamma/delta T cells, a subset of CD8+ T cells, and other immune cells and tissues (1). Additionally, 2B4 is considered a marker for CD8+ T cell exhaustion in chronic infection and increased expression has been observed in diseases including lymphocytic choriomeningitis virus (LCMV) and hepatitis B virus (HBV) (2). Human 2B4 protein is 370 amino acids (aa) in length with a theoretical molecular weight (MW) of 41 kDa (3). However, 2B4 contains multiple glycosylation sites that increases its observed MW to ~70 kDa (3). The 2B4/CD244 protein consists of a 21 aa signal peptide followed by an extracellular domain (ECD) with two Ig-like domains, a single transmembrane domain, and four cytoplasmic immunoreceptor tyrosine-based switch motifs (ITSMs) (3).

The 2B4 receptor has both activating and inhibitory signaling properties (1,4). 2B4's ligand is CD48, which is expressed on hematopoietic cells (4, 5). When bound to CD48, subsequent phosphorylation of the ITSMs occurs, followed by recruitment of SH-2-containing adaptor protein (SAP) and Fyn, leading to phosphorylation of PLCgamma and Vav1, and resulting in NK cell activation (4). In the absence of SAP, Src homology region 2-containing protein tyrosine phosphatases (SHP) get recruited and inhibits NK cell activation (4). In the context of infection such as Epstein-Barr virus (EBV), when lacking SAP, 2B4 has been shown to have an inhibitory function via impaired T cell cytotoxicity against EBV-infected cells (6). While 2B4 has a role in eliciting CD8+ T cell response, overexpression of 2B4 can lead to T cell exhaustion in states of chronic infection (6). Studies have suggested that the antibody blockade of the 2B4-CD48 interaction might enhance T cell and NK cell function, however given both the activating and inhibitory properties of the 2B4 receptor, more research needs to be done to prevent unwanted effects (6).

References

1. Buller CW, Mathew PA, Mathew SO. Roles of NK Cell Receptors 2B4 (CD244), CS1 (CD319), and LLT1 (CLEC2D) in Cancer. Cancers (Basel). 2020 Jul 1;12(7):1755. https://doi.org/10.3390/cancers12071755

2. Catakovic K, Klieser E, Neureiter D, Geisberger R. T cell exhaustion: from pathophysiological basics to tumor immunotherapy. Cell Commun Signal. 2017;15(1):1. https://doi.org/10.1186/s12964-016-0160-z

3. Uniprot (Q9BZW8)

4. Chen Y, Lu D, Churov A, Fu R. Research Progress on NK Cell Receptors and Their Signaling Pathways. Mediators Inflamm. 2020 Jul 24;2020:6437057. https://doi.org/10.1155/2020/6437057.

5. Pahima H, Puzzovio PG, Levi-Schaffer F. 2B4 and CD48: A powerful couple of the immune system. Clin Immunol. 2019 Jul;204:64-68. https://doi.org/10.1016/j.clim.2018.10.014.

6. Waggoner SN, Kumar V. Evolving role of 2B4/CD244 in T and NK cell responses during virus infection. Front Immunol. 2012;3:377. https://doi.org/10.3389/fimmu.2012.00377

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.

Bioinformatics

• Gene Symbol: CD244