Name
C3, Mouse, mAb 11H9
Catalog nr
HM1045
(lot number and expiry date are indicated on the label)
Short description
The monoclonal antibody 11H9 recognizes mouse complement component C3 and its activation products, C3b, iC3b, C3d and C3dg. The complement system is an important part of the humoral response in innate immunity, consisting of three different pathways. The third complement component, C3, is central to the classical, alternative and lectin pathways of complement activation. Activation products ...
Size
100 µg
Application
F
,
FC
,
IA
,
IF
,
IP
,
P
,
W
Technical datasheet
Description
The monoclonal antibody 11H9 recognizes mouse complement component C3 and its activation products, C3b, iC3b, C3d and C3dg. The complement system is an important part of the humoral response in innate immunity, consisting of three different pathways. The third complement component, C3, is central to the classical, alternative and lectin pathways of complement activation. Activation products of the complement cascade contain neo-epitopes that are not present in the individual native components. The complement factor C3 consists of an alpha- and a beta-chain.
The synthesis of C3 is tissue-specific and is modulated in response to a variety of stimulatory agents. C3 is the most abundant protein of the complement system with serum protein levels of about 1.3 mg/ml. An inherited deficiency of C3 predisposes the person to frequent assaults of bacterial infections. In ulcerative colitis, and idiopathic chronic inflammatory bowel disease, the deposition of C3 in the diseased mucosa has been reported.
Proteolysis by certain enzymes results in the cleavage of C3 into C3a and C3b. C3b becomes attached to immune complexes and is further cleaved into iC3b, C3c, C3dg and C3f. The monoclonal antibody 11H9 recognizes both intact C3 and its cleaved products C3b, iC3b, C3d and C3dg. These activation products are present in acute as well as chronic inflammatory conditions. In chronic inflammatory condition, primarily the C3dg product resides at the place of inflammation (C3c being cleared) which is recognized by antibody 11H9. The chronic processing/activation of C3 is taking place at a lower level, which would reduce detection of the C3 fragments C3b, iC3b, and C3c.
The synthesis of C3 is tissue-specific and is modulated in response to a variety of stimulatory agents. C3 is the most abundant protein of the complement system with serum protein levels of about 1.3 mg/ml. An inherited deficiency of C3 predisposes the person to frequent assaults of bacterial infections. In ulcerative colitis, and idiopathic chronic inflammatory bowel disease, the deposition of C3 in the diseased mucosa has been reported.
Proteolysis by certain enzymes results in the cleavage of C3 into C3a and C3b. C3b becomes attached to immune complexes and is further cleaved into iC3b, C3c, C3dg and C3f. The monoclonal antibody 11H9 recognizes both intact C3 and its cleaved products C3b, iC3b, C3d and C3dg. These activation products are present in acute as well as chronic inflammatory conditions. In chronic inflammatory condition, primarily the C3dg product resides at the place of inflammation (C3c being cleared) which is recognized by antibody 11H9. The chronic processing/activation of C3 is taking place at a lower level, which would reduce detection of the C3 fragments C3b, iC3b, and C3c.
Immunogen
C57BL/6 thymocytes saturated with rat anti-Thy-1 monoclonal antibody of IgG2b subclass (RmT1)
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.02% sodium azide and 0.1% bovine serum albumin.
Species
Rat IgG2a
Application
|
|
F1 |
FC |
FS |
IA2 |
IF3 |
IP |
P3 |
W1 |
|
Yes |
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● |
|
● |
● |
● |
● |
● |
|
No |
|
|
|
|
|
|
|
|
|
N.D. |
|
|
● |
|
|
|
|
|
Application IA and W have been tested by Hycult Biotech. Application FC and IP are based on personal communication.
Application notes
IA: Antibody 11H9 was used as coating 0.5 µg/well in PBS.
W: A reduced sample treatment and SDS-PAGE was used. The band sizes are ~110-115 kDa (α-chain), ~80 and 70 kDa (Ref.1).
F: Tissue sections were fixed in acetone. Antigen retrieval was performed using 50% formic acid for 5 min or microwaved (700 W) in antigen unmasking solution (Ref 1, 3).
IF: Fixation in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4 µm. Pretreated with 3% hydrogen peroxide for 20 min to quench endogenous peroxidases. Microwaved in antigen unmasking solution for 2-5 minutes as antigen retrieval. (Ref.3).
P: free-floating brain tissue pretreated with 50% formic acid for 5 min. or microwaved (700W) in antigen unmasking solution for 2-5min. Blocking endogenous persoxidase by treatment with 3% H2O2 in pBS 20 min at 25°C. Blocking 2hrs, 2% BSA in PBS.
C3 protein fragments deposited on kidney cells of MPL-lpr mouse. Staining with antibody 11H9 (Cat.# HM1045). Glomerular staining pattern.
W: A reduced sample treatment and SDS-PAGE was used. The band sizes are ~110-115 kDa (α-chain), ~80 and 70 kDa (Ref.1).
F: Tissue sections were fixed in acetone. Antigen retrieval was performed using 50% formic acid for 5 min or microwaved (700 W) in antigen unmasking solution (Ref 1, 3).
IF: Fixation in 4% paraformaldehyde in PBS pH 7.4. Vibratome sections of 4 µm. Pretreated with 3% hydrogen peroxide for 20 min to quench endogenous peroxidases. Microwaved in antigen unmasking solution for 2-5 minutes as antigen retrieval. (Ref.3).
P: free-floating brain tissue pretreated with 50% formic acid for 5 min. or microwaved (700W) in antigen unmasking solution for 2-5min. Blocking endogenous persoxidase by treatment with 3% H2O2 in pBS 20 min at 25°C. Blocking 2hrs, 2% BSA in PBS.
C3 protein fragments deposited on kidney cells of MPL-lpr mouse. Staining with antibody 11H9 (Cat.# HM1045). Glomerular staining pattern.
Use
For Western blotting, flow cytometry and immunohistology 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.
Aliases
Complement component C3
Positive control
MPL-lpr kidney cells
F: As positive control brain sections of APPQ mice were used and as negative control isotype matched material
F: As positive control brain sections of APPQ mice were used and as negative control isotype matched material
Negative control
Wild type kidney cells
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Kremmer, E et al; Monoclonal antibodies to complement components without the need of their prior purification. II. Antibodies to mouse C3 and C4. Hybridoma 1990, 9: 309
2. Pan, W et al; CR3 (CD11b/CD18) is the major macrophage receptor for IgM antibody-mediated phagocytosis of African trypanosomes: Diverse effect on subsequent synthesis of tumor necrosis factor a and nitric oxide. Microb Infect 2006, 8: 1209
3. Zhou, J et al; Complement C3 and C4 expression in C1q sufficient and deficient mouse models of Alzheimer's disease. J Neurochem 2008, 106: 2080
2. Pan, W et al; CR3 (CD11b/CD18) is the major macrophage receptor for IgM antibody-mediated phagocytosis of African trypanosomes: Diverse effect on subsequent synthesis of tumor necrosis factor a and nitric oxide. Microb Infect 2006, 8: 1209
3. Zhou, J et al; Complement C3 and C4 expression in C1q sufficient and deficient mouse models of Alzheimer's disease. J Neurochem 2008, 106: 2080
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
| HM1044 | C1q, Mouse, mAb 7H8 |
| HM1046 | C4, Mouse, mAb 16D2 |
| HP8012 | C3, Mouse, pAb |
References
1. Kremmer, E et al; Monoclonal antibodies to complement components without the need of their prior purification. II. Antibodies to mouse C3 and C4. Hybridoma 1990, 9: 309
2. Pan, W et al; CR3 (CD11b/CD18) is the major macrophage receptor for IgM antibody-mediated phagocytosis of African trypanosomes: Diverse effect on subsequent synthesis of tumor necrosis factor a and nitric oxide. Microb Infect 2006, 8: 1209
3. Zhou, J et al; Complement C3 and C4 expression in C1q sufficient and deficient mouse models of Alzheimer's disease. J Neurochem 2008, 106: 2080
2. Pan, W et al; CR3 (CD11b/CD18) is the major macrophage receptor for IgM antibody-mediated phagocytosis of African trypanosomes: Diverse effect on subsequent synthesis of tumor necrosis factor a and nitric oxide. Microb Infect 2006, 8: 1209
3. Zhou, J et al; Complement C3 and C4 expression in C1q sufficient and deficient mouse models of Alzheimer's disease. J Neurochem 2008, 106: 2080
Protocols
Protocols
Protocols
Protocols
Protocols
| Related products | Cat # | |
|---|---|---|
| C4, Mouse, mAb 16D2 | HM1046 | |
| C3/C3b/iC3b/C3c, Mouse, mAb 3/26 | HM1078 | |
| C3b/iC3b/C3c, Mouse, mAb 2/11 | HM1065 | |
| C1q, Mouse, mAb 7H8 | HM1044 | |
| C1q, Mouse, mAb JL-1 | HM1096 | |
| C3, Mouse, pAb | HP8012 | |