Technical Data

C7850-18K
Clone Type
Monoclonal
Host
Mouse
Isotype
IgG
Clone Number
12D11
Grade
Affinity Purified
Applications
E FC IHC WB
Crossreactivity
Bo Hu Mk Po
Shipping Temp
Blue Ice
Storage Temp
-20°C
Mouse Anti-Complement C4d

Complement factor C4 (MW 41kD), formerly known as Gg protein, consists of an alpha-, beta- and gamma-chain. The classical pathway of complement and the mannose binding lectin (MBL) activation pathway converge at C4. Activated C1, MASP-1 and MASP-2 cleave C4 resulting in the formation of C4a and C4b. The latter can be cleaved by factor I resulting in C4c and C4d, in which step all functional sites are lost. The C4d activation fragment of C4 is an excellent marker for classical complement pathway and MBL pathway activation. The thioester formed between the side chains of Cys1010 and Gln1013 within the C4d region of the alpha-chain mediates covalent attachment to the target surface bearing activated forms of C1s or MASP. Furthermore, C4d is highly homologous to C3d with over 35% shared amino acid sequence. In a number of diseases such as rheumatoid arthritis (RA), hereditary angioedema (HAE), systemic lupus erythematosus (SLE) and chronic urticaria with hypercomplementemia levels of C4d are significantly elevated in serum or plasma. C4d levels may also be elevated in plasma from patients with a variety of humoral autoimmune diseases in which complement activation is known to occur. Deposition of C4d in peritubular capillaries has been shown to be a sensitive marker for antibody-mediated (humoral) rejection in renal transplant biopsies.

Applications
Suitable for use in ELISA, Western Blot, Flow Cytometry and Immunohistochemistry. Other applications have not been tested.
Recommended Dilutions
Western Blot: 1:50 Immunohistochemistry (Frozen): 1:50 Permeabilization was done in cold acetone with 0.5% hydrogen peroxide for 10 min, after drying and washing, antibodies were incubated for 30 min. Flow Cytometry: Pneumococci were incubated with 25ul of a 1:10 diluted C7850-18K, incubated for 30 minutes and washed 2 times, followed by 30 minutes incubation on ice of a 1:100 diluted IgG (PE) goat-anti-mouse on ice in the dark and washed 2 times. Optimal dilutions to be determined by the researcher.
Storage and Stability
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Form
Supplied as as liquid in PBS, 0.1% BSA, 0.02% sodium azide.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes C4, C4b and C4d. Recognizes an epitope on the alpha 2 chain of C4d. Species Crossreactivity: human, porcine, bovine, baboon and Rhesus monkey.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Zwirner, J et al; Classical pathway of complement activation in normal and diseased human glomeruli. Kidney Int 1989, 36: 1069.2. Zwirner, J et al; Classical pathway of complement activation in mammalian kidneys. Immunology 1993, 80: 162. 3. Feucht, H et al; Capillary deposition of C4d complement fragment and early renal graft loss. Kidney Int 1993, 43: 1333.
USBio References
No references available
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