The antiserum does not cross-react with any other human plasma proteins as tested in gel-diffusion techniques. Inter-species cross-reactivity is a normal feature of antibodies to plasma proteins, since homologous proteins of different species frequently share antigenic determinants. of this antiserum has not been tested in detail, however in double radial immunodiffusion a reaction with Rhesus monkey has been observed.
Characteristics
Precipitating polyclonal rabbit antiserum to human coagulation factor VII
Purification
Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required, to eliminate antibodies reacting with other human serum proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum.
Immunogen
Plasma factor VII is ea vitamin K-dependent glycoprotein (MW 63,000) with an electrophoresis mobility in the beta region. It circulates in plasma in a semi-active form, even in the absence of tissue factor. Then procoagulant activity of FVII can be destroyed by heating to 56ºC. It is unstable below pH 3 and above pH 9. It survives he clotting process and its presence can be demonstrated in serum. After isolation its molecular weight was 44,700. Factor VII is a serine protease depending on a lipid cofactor and can be activated to FVIIa by factors IXa, Xa and XIIIa, thereby linking the intrinsic and extrinsic coagulation systems since FXII and FIX can be activated by kallikrein. The normal adult plasma FVII level is about 0.5 to 1.0 µ g/ml. In normal newborn infants the average level is about 50% of the adult concentration. A deficiency of FVII, congenital or acquired, results in a bleeding disorder. The congenital form is rare but the acquired form is commonly seen in association with a deficiency of FII, FIX and FX in liver disease and in patients taking coumarin-type anticoagulant drugs. Both procoagulant activity and FVII-related antigen are depressed. In haemorrhagic disease of the newborn, procoagulant activity is reduced but not the level of the FVII antigen. A similar discrepancy may be seen in congenital deficiencies but in other types FVII antigen will be severely reduced as a result of genetic suppression of synthesis capacity. If still present, the FVII molecules appear to be biologically defective. Heterozygote carriers can now be detected. FVII procoagulant activity and related antigen levels have been shown to correlate directly with the plasma triglyceride concentration. This makes FVII a risk factor for myocardial infarction, immediately following an acute myocardial infarction patients have increased plasma FVII procoagulant activity. FVII is purified from plasma and Freund’s complete adjuvant is used in the first step of the immunization procedure.
F7
Reactivity: Human
WB
Host: Rabbit
Polyclonal
RB54990
unconjugated
Application Notes
In precipitating techniques as electroimmunodiffusion, immunoelectrophoresis, single and double radial immuno-diffusion (Mancini, Ouchterlony), bidimensional electrophoresis and neutralization assay. The presence of non-precipitating antibodies has not been assayed. If used in more sensitive test procedures or as catching or detection antibody in solid phase immunoassays specificity controls should always be include. Plasma samples and all assay components must contain EDTA to stabilize the proteins.
Restrictions
For Research Use only
Format
Lyophilized
Concentration
Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal rabbit serum. No foreign proteins added. Antibody titre: The amount of Factor VII precipitated by 1 ml antiserum is approximately 100 U. One Unit of Factor VII
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C/-20 °C
Storage Comment
The lyophilized antiserum is shipped at ambient temperature and may be stored at +4°C, prolonged stora ge at or below -20°C. Reconstitute the lyophilized antiserum by add ing 1 ml sterile distilled water. Dilutions may be prepared by adding phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the antiserum. Diluted antiserum should be stored at +4°C, not ref rozen, and preferably used the same day.
The defined antibody reactivity is restricted to Factor VII. To demonstrate the presence of FVII in normal plasma or serum by gel-immunodiffusion or immunoelectrophoresis a concentrate must be prepared, because the normal concentration is below the detection limits of these techniques. The precipitation obtained with the concentrate shows a reaction of identity with that obtained with the purified factor. FVII shows micro-heterogeneity. The antiserum also reacts with FVII molecular variants and with abnormal, molecules (PIVKA VII)