The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Vaccinia virus is an Orthopoxvirus, containing double stranded DNA. Fusion protein plays an important role in the entry of enveloped virus into cells. As vaccinia virus has a wide host range, it is conceivable that certain cellular components that are ubiquitously expressed on the cell mediate virus infection. The study of the entry process, attachment, fusion and the proteins and receptors involved is complex. During vaccinia virus infection, the fusion process is attributed to the action of the 14KDa protein (A27L). The N terminus of this protein recognises heparan sulfate on the cell surface. It interacts with the negative charges of sulfates of glycosaminoglycans (GAGs). Therefore, antibodies that recognize this 14KDa protein are able to neutralize vaccinia virus infection and enable identification other viral and cellular proteins which participate in the vaccinia virus entry process.
ELISA - Vaccinia Virus antibody (ab26853)
ELISA method: Antigen is coated on EIA strips at 1µg per well. Add 200µl blocking buffer then wash wells with PBST. Antiserum or peptide specific purified antibody is diluted in series as 102~106 folds. Incubate antibody for 1 hour. Wash unbound antibodies and add HRP-conjugated anti-rabbit IgG. Wash plates and add substrate to develop color for 5 minutes. Read absorbance at 650nm. Amount of colour is directly proportional to amount of antibodies. Antibody titer is defined as >0.1 of absorbance of antiserum minus pre-immune serum. Titer ~ 1/437000.
has not yet been referenced specifically in any publications.