Technical datasheet
Description
The monoclonal antibody 16D2 recognizes mouse complement factor C4, formerly known as Gg protein, which consists of an alpha-, beta-, and gamma-chain. The classical pathway of complement and the Mannose binding lectin activation pathway converge at C4. C1s, MASP-1 and MASP-2 cleave C4 resulting in the formation of C4a and C4b. Subsequently, C4b can be cleaved to C4c and C4d by other serum enzymes. The monoclonal antibody 16D2 reacts with intact C4, C4b and C4d.
C4 is an acute phase protein that is produced by hepatocytes, monocytes and intestinal epithelial cells and can be used in experimental animals as a marker for activation of the classical complement pathway. Recent studies have demonstrated an association between graft rejection and C4d deposition in a mouse model for cardiac transplantation.
Cross Reactivity
| Cross reactant |
Reactivity |
| C4b |
Yes |
| C4d |
Yes |
Immunogen
Thymocytes decorated with Thy-1 antibody and complement components
Formulation
1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin and 0.02% sodium azide.
Species
Rat IgG2a
Application
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F1
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FC
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FS
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IA2,3
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IF2
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IP1
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P
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W1,4
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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
W: A non-reduced sample treatment and SDS-Page was used. The band size is 200x kDa (Ref.4).
IHC-F: Tissue sections were fixed in ice-cold acetone and blocked with 2% BSA, FCS and PBS. (Ref.1).
Use
For immunohistochemistry and Western blotting, 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 4
Positive control
T-cell regions of mouse spleen
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least one year. The exact expiry date is indicated on the label.
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. Gadjeva, M et al; Macrophage-derived complement component C4 can restore humoral immunity in C4-deficient mice. J Immunol 2002, 169: 5489
3. Paul E et al; Anti-DNA autoreactivity in C4-deficient mice. Eur J Immunol 2002, 32: 2672
4. Murata, K et al; Synergistic deposition of C4d by complement-activating and non-activating antibodies in cardiac transplants. Am J Transplant 2007, 7: 2605
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 from the use or derivation of this product.
Also available
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. Gadjeva, M et al; Macrophage-derived complement component C4 can restore humoral immunity in C4-deficient mice. J Immunol 2002, 169: 5489
3. Paul E et al; Anti-DNA autoreactivity in C4-deficient mice. Eur J Immunol 2002, 32: 2672
4. Murata, K et al; Synergistic deposition of C4d by complement-activating and non-activating antibodies in cardiac transplants. Am J Transplant 2007, 7: 2605