Name
C5, Mouse, mAb BB5.1
Catalog nr
HM1073
(lot number and expiry date are indicated on the label)
Short description
The monoclonal antibody BB5.1 binds the fifth component of mouse complement (C5). The complement system is a group of plasma and cell membrane proteins that play a key role in the immune system. All three pathways (classical, alternative & lectin) lead to the cleavage of C3 and eventually the formation of the cytolytic membrane attack complex C5b-9. If the activation cascade is allowed to proceed beyond ...
Size
100 µg
Application
F
,
FS
,
IA
,
IP
,
P
Technical datasheet
Description
The monoclonal antibody BB5.1 binds the fifth component of mouse complement (C5). The complement system is a group of plasma and cell membrane proteins that play a key role in the immune system. All three pathways (classical, alternative & lectin) lead to the cleavage of C3 and eventually the formation of the cytolytic membrane attack complex C5b-9. If the activation cascade is allowed to proceed beyond the cleaving of C3 into C3a and C3b, an additional C3b molecule binds to the C3 convertases. This generates the C5 convertase (C3bBbC3b for the alternative pathway and C4bC2bC3b for the classical and lectin pathways). C5 convertase cleaves C5, releases the potent anaphylactic peptide C5a and generates C5b. C5b can initiate the terminal pathway , which recruits the components C6, C7, C8 and C9 to the surface of the target and inserts the C9 complex as a pore (termed the terminal complement complex) into the membrane. C5a is the most potent anaphylatoxin and a powerful chemotaxin for neutrophils and monocytes, with the ability to promote margination, extravasation, and activation of these cells. Thus, blocking C5 may be required for optimal inhibition of the inflammatory response. At the same time, inhibition of the complement cascade at C5 does not impair the generation of C3b through the classical and alternative pathways, preserving C3b-mediated opsonization of pathogenic microorganisms as well as opsonization and solubilization of immune complexes.
C5 is synthesised in the liver as a single polypeptide chain and is present in serum in a concentration of 50-80 µg/ml. Before secretion the molecule is glycosylated and secreted into plasma as a 190 kDa glycoprotein consisting of a disulphide linked alpha-chain (111 kDa) and beta-chain (75 kDa). Monoclonal antibody BB5.1 has been shown to precipitate the two chains of C5 from normal mouse serum and inhibited C5-dependent hemolysis in a functional complement test. Furthermore, BB5.1 administration completely inhibits terminal complement activity in murine models for antibody-mediated rejection (AMR) during heart and kidney transplantation. In another mouse model, both pre-treatment as well as intervention with monoclonal antibody BB5.1 attenuates disease development during anti-MPO IgG-induced glomerulonephritis. Blockage of C5 activation by BB5.1 protects against renal ischemia-reperfusion injury by inhibition of late apoptosis and inflammation. In Lupus disease, combination therapy of anti-IL-10/anti-C5 (BB5.1) can both prevent and reduce the effect of the humoral immune response.
C5 is synthesised in the liver as a single polypeptide chain and is present in serum in a concentration of 50-80 µg/ml. Before secretion the molecule is glycosylated and secreted into plasma as a 190 kDa glycoprotein consisting of a disulphide linked alpha-chain (111 kDa) and beta-chain (75 kDa). Monoclonal antibody BB5.1 has been shown to precipitate the two chains of C5 from normal mouse serum and inhibited C5-dependent hemolysis in a functional complement test. Furthermore, BB5.1 administration completely inhibits terminal complement activity in murine models for antibody-mediated rejection (AMR) during heart and kidney transplantation. In another mouse model, both pre-treatment as well as intervention with monoclonal antibody BB5.1 attenuates disease development during anti-MPO IgG-induced glomerulonephritis. Blockage of C5 activation by BB5.1 protects against renal ischemia-reperfusion injury by inhibition of late apoptosis and inflammation. In Lupus disease, combination therapy of anti-IL-10/anti-C5 (BB5.1) can both prevent and reduce the effect of the humoral immune response.
Cross Reactivity
| Cross reactant | Reactivity |
| Rat | Yes |
Immunogen
Purified C5
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.
Species
Mouse IgG1
Application
|
|
F3 |
FC |
FS 2,4,5,7 |
IA 1,6 |
IF |
IP1 |
P4 |
W |
|
Yes |
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● |
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● |
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|
No |
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N.D. |
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Application notes
FS: ex vivo: 50µg mAb / ml serum; in vivo:BALB/c mice 40mg mAb/kg/day, 0-2 days followed by twice a week
IA: Wells were coated with 2-4µg purified BB5.1 in PBS as coat antibody for use in ELISA.
IP: Mouse serum was pre-adsorbed with αmouse-Ig affinity gel and prot-A-sepharose for 30min at RT. Then C5 was precipitated with BB5.1 (10µg mAb/ml followed by prot-A sepharose for 1h). Precipitated samples were analysed on 7% SDS/PAGE
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IA: Wells were coated with 2-4µg purified BB5.1 in PBS as coat antibody for use in ELISA.
IP: Mouse serum was pre-adsorbed with αmouse-Ig affinity gel and prot-A-sepharose for 30min at RT. Then C5 was precipitated with BB5.1 (10µg mAb/ml followed by prot-A sepharose for 1h). Precipitated samples were analysed on 7% SDS/PAGE
Use
For 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. For functional studies, in vitro dilutions have to be optimized in user's experimental setting.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.
References
1. Frei, Y et al; Generation of a monoclonal antibody to mouse C5 application in an ELISA assay for detection of anti-C5 antibodies. Mol Cell Probes 1987, 1: 141
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
6. Nilsson, K et al. Enhanced susceptibility to low-dose collagen-induced arthritis in CR1/2-deficient female mice – possible role of estrogen on CR1 expression. Faseb 2009, 23: 2450
7. Copland, D et al. Systemic and local anti-C5 therapy reduces the disease severity in experimental autoimmune uveoretinitis. Clin exp immunol 2010, 159:303
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
6. Nilsson, K et al. Enhanced susceptibility to low-dose collagen-induced arthritis in CR1/2-deficient female mice – possible role of estrogen on CR1 expression. Faseb 2009, 23: 2450
7. Copland, D et al. Systemic and local anti-C5 therapy reduces the disease severity in experimental autoimmune uveoretinitis. Clin exp immunol 2010, 159:303
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 or derivation of this product.
Also available
| HM1096 | C1q, Mouse, mAb JL-1 |
| HM1045 | C3, Mouse, mAb 11H9 |
| HM1046 | C4, Mouse, mAb 16D2 |
| HP8013 | C5, Mouse, pAb |
| HM1051 | MPO, Mouse, mAb 8F4 |
References
1. Frei, Y et al; Generation of a monoclonal antibody to mouse C5 application in an ELISA assay for detection of anti-C5 antibodies. Mol Cell Probes 1987, 1: 141
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
6. Nilsson, K et al. Enhanced susceptibility to low-dose collagen-induced arthritis in CR1/2-deficient female mice – possible role of estrogen on CR1 expression. Faseb 2009, 23: 2450
7. Copland, D et al. Systemic and local anti-C5 therapy reduces the disease severity in experimental autoimmune uveoretinitis. Clin exp immunol 2010, 159:303
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
6. Nilsson, K et al. Enhanced susceptibility to low-dose collagen-induced arthritis in CR1/2-deficient female mice – possible role of estrogen on CR1 expression. Faseb 2009, 23: 2450
7. Copland, D et al. Systemic and local anti-C5 therapy reduces the disease severity in experimental autoimmune uveoretinitis. Clin exp immunol 2010, 159:303
Scientific info
Anti-Mouse C5, Clone BB5.1 inhibits membrane attack complex formation
Special features:
• monoclonal antibody BB5.1 specific for the fifth component of mouse complement (C5)
• the complement activation pathway via activated C3 to the cleavage of C5 into C5a and C5b
• monoclonal antibody BB5.1 prevents cleavage of C5, thereby preventing C5a generation and MAC formation
• BB5.1 administration completely inhibits terminal complement activity in murine models for antibody-mediated rejection (AMR) during heart and kidney transplantation
• BB5.1 attenuates disease development during anti-MPO IgG-induced glomerulonephritis
• inhibition of C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation
• effective in combination therapy of anti-IL-10/ anti-C5 to prevent or reduce the effect of the humoral immune response in Lupus disease
• quantity of 100 µg/ml available for in vitro research purposes
• bulk quantities available of 5 mg and 10 mg
Selected reading
1. Frei, Y et al; Generation of a monoclonal antibody to mouse C5 application in an ELISA assay for detection of anti-C5 antibodies. Mol Cell Probes 1987, 1: 141
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Rother, R et al; C5 blockade with conventional immunosuppression induces long-term graft survival in presensitized recipients. Am J Transplant 2008, 8(6): 1129
6. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
7. de Vries, B et al: Inhibition of C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation. Transplantation 2003, 75:375
Special features:
• monoclonal antibody BB5.1 specific for the fifth component of mouse complement (C5)
• the complement activation pathway via activated C3 to the cleavage of C5 into C5a and C5b
• monoclonal antibody BB5.1 prevents cleavage of C5, thereby preventing C5a generation and MAC formation
• BB5.1 administration completely inhibits terminal complement activity in murine models for antibody-mediated rejection (AMR) during heart and kidney transplantation
• BB5.1 attenuates disease development during anti-MPO IgG-induced glomerulonephritis
• inhibition of C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation
• effective in combination therapy of anti-IL-10/ anti-C5 to prevent or reduce the effect of the humoral immune response in Lupus disease
• quantity of 100 µg/ml available for in vitro research purposes
• bulk quantities available of 5 mg and 10 mg
Selected reading
1. Frei, Y et al; Generation of a monoclonal antibody to mouse C5 application in an ELISA assay for detection of anti-C5 antibodies. Mol Cell Probes 1987, 1: 141
2. Thurman, J et al; C3a is required for the production of CXC chemokines by tubular epithelial cells after renal ischemia/reperfusion. J Immunol 2007, 178: 1819
3. Mihai, S et al; The alternative pathway of complement activation is critical for blister induction in experimental epidermolysis bullosa acquisita. J Immunol 2007, 178: 6514
4. Wang, H et al; Inhibition of terminal complement components in presensitized transplant recipients prevents antibody-mediated rejection leading to long-term graft survival and accommodation. J Immunol 2007, 179: 4451
5. Rother, R et al; C5 blockade with conventional immunosuppression induces long-term graft survival in presensitized recipients. Am J Transplant 2008, 8(6): 1129
6. Huugen, D et al; Inhibition of complement factor C5 protects against anti-myeloperoxidase antibody-mediated glomerulonephritis in mice. Kidney Int 2007, 71: 646
7. de Vries, B et al: Inhibition of C5 protects against renal ischemia-reperfusion injury: inhibition of late apoptosis and inflammation. Transplantation 2003, 75:375
Protocols
Protocols
Protocols
| Related products | Cat # | |
|---|---|---|
| C3, Mouse, mAb 11H9 | HM1045 | |
| C4, Mouse, mAb 16D2 | HM1046 | |
| MPO, Mouse, mAb 8F4 | HM1051 | |
| C1q, Mouse, mAb JL-1 | HM1096 | |
| C5, Mouse, pAb | HP8013 | |