Human SP-D Antibody

Catalog # Availability Size / Price Qty
MAB19201-100
MAB19201-SP
Human  SP‑D ELISA Standard Curve.
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Human SP-D Antibody Summary

Species Reactivity
Human
Specificity
Detects human SP-D in direct ELISAs.
Source
Monoclonal Mouse IgG2B Clone # 292215
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human SP-D
Ala21-Phe375
Accession # P35247.2
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
ELISA

This antibody functions as an ELISA capture antibody when paired with Mouse Anti-Human SP‑D Monoclonal Antibody (Catalog # MAB1920).

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human SP-D DuoSet ELISA Kit (Catalog # DY1920) for convenient development of a sandwich ELISA or the Human SP-D Quantikine ELISA Kit (Catalog # DSFPD0) for a complete optimized ELISA.

 

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.

Data Example

Human SP‑D ELISA Standard Curve. Recombinant Human SP‑D protein was serially diluted 2-fold and captured by Mouse Anti-Human SP‑D Monoclonal Antibody (Catalog # MAB19201) coated on a Clear Polystyrene Microplate (Catalog # DY990). Mouse Anti-Human SP‑D Monoclonal Antibody (Catalog # MAB1920) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

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Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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: SP-D

SP-D (surfactant protein-D; also PSP-D) is a 43 kDa member of the collectin family of innate immune modulators. It is constitutively secreted by alveolar lining cells and epithelium associated with tubular structures. Its principal components consist of a collagen-like region and a C-terminal carbohydrate recognition domain (CRD), a structure that further places it in a subset of an expanded group of proteins termed defense collagens (1-4). Human SP-D is synthesized as a 375 amino acid (aa) precursor. It contains a 20 aa signal sequence and a 355 aa mature region. The mature molecule is characterized by the presence of a 25 aa N-terminal linking-region, a 177 aa hydroxyproline and hydroxylysine collagen-like domain, a 46 aa coiled-coil segment, and a 106 aa, C-terminal collectin-like C-type lectin domain (CRD) (5, 6). Two additional, potential isoforms exist. One shows a 13 aa N-terminal extension, while the other combines the N-terminal extension with a deletion of aa’s 206-375. Mature human SP-D shares 75% and 78% aa identity with mouse and pig SP-D, respectively. Monomeric SP-D is unusual (3). The basic form of SP-D is that of a glycosylated, disulfide-linked 150 kDa trimer that generates an alpha -helical coiled-coil structure linked to a “head” of three symmetrical CRDs (4, 7). Each CRD recognizes the hydroxides of one monosaccharide (4). Trimerization allows for the discrimination of monosaccharide patterns specific to microbial pathogens (7). Typically, SP-D forms a higher-order 620 kDa, X-shaped dodecamer through disulfide bonds associated with the N-terminus (8). This allows for even finer discrimination of self vs. nonself carbohydrate patterns, and facilitates binding to complex antigens (8, 9). One polymorphism, a Met11-Thr11 transition in human, apparently precludes the formation of oligomers, potentially affecting the ability of affected individuals to interact with microorganisms (9, 10). Finally, SP-D is known to bind both SIRP alpha and the calreticulin/CD91 complex on macrophages. When the ratio of antigen/pathogen to available CRDs is low, antigen can be bound without occupying all available CRDs. The free CRDs will bind to SIRP alpha, generating a signal that downmodulates the inflammatory response. When virtually all CRDs are occupied by ligand, however, free CRDs are not available for SIRP alpha binding. Instead, the dodecamer is depicted to undergo a structural rearrangement, exposing the N-termini of all four linked trimers. This exposed terminus is known to bind to the calreticulin/CD91 complex, an event that initiates inflammation. Thus, it would appear that SP-D allows for a graded response to environmental challenge. SP-D provides a mechanism for the clearance of small antigenic insults without the need for a damaging inflammatory response (3).

References
  1. Holmskov, U. et. al. (2003) Annu. Rev. Immunol. 21:547.
  2. Kishore, U. et. al. (2006) Mol. Immunol. 43:1293.
  3. Hartl, D. and M. Griese (2006) Eur. J. Clin. Invest. 36:423.
  4. Sim, R.B. et. al. (2006) Novartis Found Symp. 279:170.
  5. Rust, K. et. al. (1991) Arch. Biochem. Biophys. 290:116.
  6. Lu, J. et. al. (1992) Biochem. J. 284:795.
  7. Hakansson, K. et. al. (1999) Structure 7:225.
  8. Ohya, M. et. al. (2006) Biochemistry 45:8657.
  9. Crouch, E.C. et. al. (2006) Am. J. Respir. Cell Mol. Biol. 35:84.
  10. Leth-Larsen, R. et. al. (2005) J. Immunol. 174:1532.
Long Name
Surfactant Pulmonary Associated Protein D
Entrez Gene IDs
6441 (Human); 20390 (Mouse)
Alternate Names
COLEC7collectin-7; Collectin 7; Collectin-7; Lung surfactant protein D; PSPD; PSP-D; SFTP4; SFTP4pulmonary surfactant-associated protein D; SFTPD; SPD; SP-D; SP-Dpulmonary surfactant apoprotein; surfactant protein D; surfactant, pulmonary-associated protein D; surfactant-associated protein, pulmonary 4

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