Human MD2 Biotinylated Antibody

Catalog #: BAF1787 Datasheet / COA / SDS

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BAF1787

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Human MD2 Biotinylated Antibody Summary

Species Reactivity

Human

Specificity

Detects human MD-2 in Western blots. In Western blots, less than 1% cross-reactivity with recombinant human MD-1 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human MD-2
Glu17-Asn160
Accession # NP_001123946

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.

Label

Biotin

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human MD‑2 (Catalog # 1787-MD)

Immunocytochemistry

5-15 µg/mL

Immersion fixed U937 human histiocytic lymphoma cell line

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

Immunocytochemistry

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MD2 in U937 Human Cell Line. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it 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: MD2

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).

References