Name: GVBo (GVB without Ca++ and Mg++)
Catalog Numbers: B101 and B103
Sizes Available: 1000 mL and 250 mL
Composition: 0.1 % gelatin, 5 mM Veronal, 145 mM NaCl, 0.025 % NaN3 ,
Buffer: Sodium veronal
Preservative: Sterile filtered with 0.025% sodium azide as a bactericidal agent
Storage: +4oC Avoid freezing which causes gelatin to gel. If frozen, heat
to redissolve gelatin.
Precautions: Azide is poisonous to all living organisms.
Origin: Manufactured in the
GVBo is a basic buffer that can be used to make other traditional buffers used for complement assays. Ca++ and Mg++ may be added to GVBo to make GVB++ for classical pathway and lectin pathway assays. MgEGTA may be added to GVBo for alternative pathway assays. EDTA may be added to GVBo to prepare GVBE to inhibit complement activation. GVBo is also used for dilution of serum and other assay components in many complement assays especially in alternative pathway assays (Morgan, B.P. (2000; Dodds, A.W. and Sim, R.B. (1997)).
Buffers such as GVBo, GVB++ and GVBE need to be ordered by Friday in order to receive them the next week. They are shipped Monday afternoon by overnight courier for delivery on Tuesday or Wednesday. They can usually be used for 3 months after preparation if kept cold @ 4oC. They are shipped cold, but are not harmed at room temperature and must be warmed to 37oC for assays.
Veronal is used as the buffer because in the mid-1900s this was the only buffer for pH range 7.2-7.4 that did not chelate metal ions and did not to inhibit complement reactions as did other buffers. Sodium chloride is present to provide an isotonic environment so that cells do not lyse due to osmotic pressure. Gelatin is present to prevent loss of protein components due to adsorption onto tips or tubes during dilutions and in the assays themselves. Azide is present to prevent bacterial growth.
The concentration of gelatin in this buffer is below the concentration that forms solid gels. However, because of the gelatin is close to its gelling concentration at 4oC some strings of gelatin form during standing at this temperature. They can be redissolved easily by heating to 37oC or by brief heating in a microwave oven.
GVBo should be used to prepare Er and to wash them before their use in assays. GVBo is used to dilute all of the components of alternative pathway assays. The General Description section above describes the use of GVBo to prepare GVB++ , MgEGTA for alternative pathway assays and EDTA to prepare GVBE. GVBo is also used for dilution of serum and other assay components in many complement assays especially in alternative pathway assays.
Morgan, B.P. ed. (2000) Complement Methods and Protocols. Humana Press.
Dodds, A.W. and Sim, R.B. editors (1997).
Complement A Practical Approach (ISBN 019963539)
FOR RESEARCH USE ONLY.
NOT FOR HUMAN OR DRUG USE.
Complement Technology, Inc.