SP-D Antibody [Unconjugated]

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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

SP-D Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human SP-D
  Ala21-Phe375 (Glu22Gly)
  Accession # P35247
• Specificity: Detects human SP-D in direct ELISAs and human, mouse, and rat SP-D in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SFTPD
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Simple Western 10 ug/mL
  • Western Blot 0.2 ug/mL

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 Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• 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 SP-D Antibody [Unconjugated]

• 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

Background

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).

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.

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.

Publications for SP-D Antibody (AF1920)(5)

Publications using AF1920

• A Zoued, H Zhang, T Zhang, RT Giorgio, CJ Kuehl, B Fakoya, B Sit, MK Waldor Proteomic analysis of the host-pathogen interface in experimental cholera Nature Chemical Biology, 2021-10-21;17(11):1199-1208. 2021-10-21 [PMID: 34675415] (IHC, Western Blot, Rabbit)
• S Yousefi, SK Sharma, D Stojkov, N Germic, S Aeschliman, MQ Ge, CH Flayer, ED Larson, IG Redai, S Zhang, CJ Koziol-Whi, K Karikó, HU Simon, A Haczku Oxidative damage of SP-D abolishes control of eosinophil extracellular DNA trap formation J. Leukoc. Biol., 2018-05-07;0(0):. 2018-05-07 [PMID: 29733456] (Western Blot, Mouse)
• Preston E. Bratcher, Amit Gaggar Factors Influencing the Measurement of Plasma/Serum Surfactant Protein D Levels by ELISA PLoS ONE 2014-11-03 [PMID: 25365324]
• Preston E. Bratcher, Amit Gaggar Characterization and Prevention of the Adsorption of Surfactant Protein D to Polypropylene PLoS ONE 2013-09-11 [PMID: 24039953]
• Bratcher , Preston, Weathington , Nathanie, Nick , Heidi J, Jackson , Patricia, Snelgrove , Robert J, Gaggar , Amit MMP-9 cleaves SP-D and abrogates its innate immune functions in vitro. PLoS ONE, 2012-07-30;7(7):e41881. 2012-07-30 [PMID: 22860023] (Human)

Bioinformatics

• Gene Symbol: SFTPD
• Uniprot:
  • P35247()