MD-2 Antibody [Biotin]

MD2 was detected in immersion fixed U937 human histiocytic lymphoma cell line using Sheep Anti-Human MD2 Biotinylated Antigen Affinity-purified Polyclonal Antibody (Catalog # BAF1787) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; NL999) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Biotin
• Concentration: LYOPH

MD-2 Antibody [Biotin] Summary

• Immunogen: E. coli-derived recombinant human MD-2
  Glu17-Asn160
  Accession # NP_001123946
• Specificity: Detects human MD-2 in Western blots. In Western blots, less than 1% cross-reactivity with recombinant human MD-1 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: LY96
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.1 ug/mL
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for MD-2 Antibody [Biotin]

• ESOP1
• ESOP-1
• LY96
• ly-96
• lymphocyte antigen 96
• MD2
• MD-2
• myeloid differentiation protein-2
• Protein MD-2

Background

MD-2, also known as lymphocyte antigen 96 and ESOP-1, is a secreted glycoprotein that shares conserved cysteine residues and significant sequence similarity (23%) with MD-1. The gene of human MD-2 encodes a 160 amino acid residue (aa) precursor protein with a 16 aa signal peptide and a 144 aa mature protein, which contains 2 N-glycosylation sites (1). Recombinant secreted MD-2 has been found to exist as disulfide-linked dimers and oligomers (2).

Both MD-1 and MD-2 are accessory molecules that associate with the extracellular leucine-rich repeats (LRR) of Toll-like receptor (TLR) family members, which are type I transmembrane receptors that regulate innate immune responses to microbial pathogens (3, 4). MD-1 binds to RP105 on B cells and macrophages to form the signaling receptor complex for lipopolysaccharide (LPS), a constituent of the outer membrane of Gram-negative bacteria. Similarly, MD-2 interacts with TLR-4 to form the heteromeric receptor that confers LPS responsiveness. MD-2 also associates with TLR-2, albeit with less avidity, to confer responsiveness to cell wall components from both Gram-positive and Gram-negative bacteria. MD-1 and MD-2 are also required for the correct targeting of the TLRs to the cell surface. Although MD-2 glycosylation is not crucial for its surface expression and interaction with TLR-4, it is required for LPS binding and signaling (5).

1. Shimazu, R. et al. (1999) J. Exp. Med. 189:1777.
2. Visintin, A. et al. (2001) Proc. Natl. Acad. Sci. USA 98:12156.
3. Nagai, Y. et al. (2002) Nature Immunology 3:667.
4. Akashi, S. et al. (2003) J. Exp. Med. 198:1035.
5. Correia, J. and R. Ulevitch (2002) J. Biol. Chem. 277:1845.

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: LY96
• Uniprot:
  • NP_001123946()