Purified, full length native human complement factor C9 protein.
ab17931 recognizes C9 in human serum diluted 1:50 in Tris buffer (20 mM Tris-base, 1 mM MgCl2, 1 mM CaCl2 and 140 mM NaCl) and incubated for 2 hours at 37°C using a human IgM coated (10 µg/mL overnight at 4°C, blocked with PBS 7.2 + 1% BSA for 1 hour) ELISA plate.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Primary antibody notesab17931 recognizes C9 in human serum diluted 1:50 in Tris buffer (20 mM Tris-base, 1 mM MgCl2, 1 mM CaCl2 and 140 mM NaCl) and incubated for 2 hours at 37°C using a human IgM coated (10 µg/mL overnight at 4°C, blocked with PBS 7.2 + 1% BSA for 1 hour) ELISA plate.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. PubMed: 20707004
Use a concentration of 2 µg/ml.
FunctionConstituent of the membrane attack complex (MAC) that plays a key role in the innate and adaptive immune response by forming pores in the plasma membrane of target cells. C9 is the pore-forming subunit of the MAC.
Involvement in diseaseDefects in C9 are a cause of complement component 9 deficiency (C9D) [MIM:613825]. A rare defect of the complement classical pathway associated with susceptibility to severe recurrent infections, predominantly by Neisseria gonorrhoeae or Neisseria meningitidis.
Sequence similaritiesBelongs to the complement C6/C7/C8/C9 family. Contains 1 EGF-like domain. Contains 1 LDL-receptor class A domain. Contains 1 MACPF domain. Contains 1 TSP type-1 domain.
Post-translational modificationsThrombin cleaves factor C9 to produce C9a and C9b. Phosphorylation sites are present in the extracelllular medium.
Cellular localizationSecreted. Cell membrane. Secreted as soluble monomer. Oligomerizes at target membranes, forming a pre-pore. A conformation change then leads to the formation of a 100 Angstrom diameter pore.
Ab17931 staining human normal liver. Staining is localized to the cytoplasm. Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer, EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
Flow Cytometry-Anti-C9 antibody (ab17931)
Overlay histogram showing HeLa cells stained with ab17931 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab17931, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.