Technical datasheet
Description
The monoclonal antibody 3/11 recognizes C3a, an activated fragment of the complement protein C3, a 190 kDa protein which is the most abundant complement protein in serum. C3 is the third component of the complement system and plays a central role in the complement cascade. The three pathways of compliment activation come together at this key molecule. Cleavage of the pivotal complement component C3 results in the generation of a set of effector molecules which have diverse biological functions. The cleavage product C3b promotes complement acrtivation and the subsequent formation of the membrane attack complex. C3a, the small (ca. 10 kDa) cleavage product possesses anaphylatoxic as well as immunoregulatory properties. It exerts its function through a specific receptor (C3aR), which belongs to the G-protein coupled receptor family. Expression of C3aR has been reported in many cell types including myeloid and non-myeloid cells. Expression of C3aR on haematopoietic stem/progenitor cells has been shown to promote the trafficking/homing of these cells to the BM. Engagement of C3aR on DCs and T cells has been shown to up-regulate these cell functions.
The monoclonal antibody 3/11 reacts specifically with the C3a anaphylatoxin, providing a tool for monitoring acute complement activation (C3a generation) in experimental mouse models.
Cross Reactivity
| Cross reactant |
Reactivity |
| C3 |
Yes |
| C3c |
No |
Immunogen
Purified mouse C3
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, and 0.1% bovine serum albumin.
Species
Rat IgG2a
Application
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F
|
FC1
|
FS4
|
IA1,3,5
|
IF
|
IP
|
P
|
W1
|
|
Yes
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No
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N.D.
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N.D.= Not Determined; F = Frozen sections; FC = Flow Cytometry; FS = Functional Studies; IA = Immuno Assays; IF = Immuno Fluorescence; IP = Immuno Precipitation; P = Paraffin sections; W = Western blot
Application notes
IA: Plates were coated with goat anti rat IgG-Fc antibody, then incubated with 1:100 hybridoma supernatant to form capture complex for C3a.
FS: Antibody 3/11 functions as an antagonist. The antibody can be used to block C3a (Ref.4).
W: A non-reduced sample treatment and 7.5% polyacrylamide gel was used. Proteins were blotted on nitrocellulose, blocked with 5% skimmed milk and incubated with 1:10 culturesupernatant for 1h at RT. The band size of the entire C3 molecule is 190 kDa (Ref.1).
Use
For Western blotting (non-reduced) dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50.
Negative control
Serum and tissue of C3-/- mice
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Mastellos, D et al; Novel monoclonal antibodies against mouse C3 interfering with complement activation: description of fine specificity and applications to various immunoassays. Mol Immunol 2004, 40: 1213
2. Markiewski, M et al; C3a and C3b activation products of the third component of complement (C3) are critical for normal liver recovery after toxic injury. J Immunol 2004, 173: 747
3. Park, J et al; Complement activation is critical to airway hyperresponsiveness after acute ozone exposure. Am J Respir Crit Care Med 2004, 169: 726
4. Nozaki, M et al; Drusen complement components C3a and C5a promote choroidal neovascularization. PNAS 2006, 103: 2328
5. Pritchard, M et al; Differential contributions of C3, C5 and decay accelerating factor to ethanol-induced fatty liver in mice. Gastroenterology 2007, 132:1117
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
References
1. Mastellos, D et al; Novel monoclonal antibodies against mouse C3 interfering with complement activation: description of fine specificity and applications to various immunoassays. Mol Immunol 2004, 40: 1213
2. Markiewski, M et al; C3a and C3b activation products of the third component of complement (C3) are critical for normal liver recovery after toxic injury. J Immunol 2004, 173: 747
3. Park, J et al; Complement activation is critical to airway hyperresponsiveness after acute ozone exposure. Am J Respir Crit Care Med 2004, 169: 726
4. Nozaki, M et al; Drusen complement components C3a and C5a promote choroidal neovascularization. PNAS 2006, 103: 2328
5. Pritchard, M et al; Differential contributions of C3, C5 and decay accelerating factor to ethanol-induced fatty liver in mice. Gastroenterology 2007, 132:1117