Human/Mouse/Rat SP-D Antibody

(1 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human SP-D in direct ELISAs and human, mouse, and rat SP-D in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human SP-D
    Ala21-Phe375 (Glu22Gly)
    Accession # P35247
  • 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
  • Western Blot
    0.2 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
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 Examples
Detection of Human, Mouse, and Rat
SP‑D by Western Blot.
Western blot shows lysates of human lung tissue, mouse lung tissue, and rat lung tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human/Mouse/Rat SP‑D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1920) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for SP‑D at approximately 43 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human and Mouse SP‑D by Simple WesternTM. Simple Western lane view shows lysates of human lung tissue and mouse lung tissue, loaded at 0.2 mg/mL. A specific band was detected for SP‑D at approximately 64/59 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat SP‑D Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1920) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
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. *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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. MMP-9 cleaves SP-D and abrogates its innate immune functions in vitro.
    Authors: Bratcher, Preston, Weathington, Nathanie, Nick, Heidi J, Jackson, Patricia, Snelgrove, Robert J, Gaggar, Amit
    PLoS ONE, 2012;7(7):e41881.
    Species: Human
    Sample Type: Recombinant Protein
Isotype Controls
Description Application Cat# Citations Images  

Normal Goat IgG Control

Ctrl AB-108-C 191  
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Secondary Antibodies
Description Application Cat# Citations Images  

Goat IgG HRP-conjugated Antibody

WB, Simple Western HAF109 19  
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Goat IgG HRP-conjugated Antibody

WB HAF017 15  
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Goat IgG (H+L) PE-conjugated Antibody

Flow F0107 4  
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Donkey Anti-Goat IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL001 9
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Donkey Anti-Goat IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL003 5
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Goat IgG (H+L) APC-conjugated Antibody

Flow F0108 6  
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Goat IgG Horseradish Peroxidase-conjugated Antibody

WB HAF019 6  
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Donkey Anti-Goat IgG Antibody

WB AF109 6
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Donkey Anti-Goat IgG Biotinylated Antibody

WB BAF109 3
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Goat IgG VisUCyte HRP Polymer

IHC VC004  
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Goat IgG (H+L) Fluorescein-conjugated Antibody

Flow F0109 2  
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Donkey Anti-Goat IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL002
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Chicken Anti-Goat IgG Biotinylated Antibody

WB BAF019
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Rabbit Anti-Goat IgG Biotinylated Antibody

WB BAF017
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Rabbit Anti-Goat IgG (H+L) Affinity Purified PAb, X Absorbed

WB, ELISA R-401-C-ABS
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Donkey Anti-Goat IgG (H+L) PerCP-conjugated Antibody

Flow F0124
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