General Antibody Information

    Rockland Immunochemicals' produces antisera in a licensed animal facility by hyper-immunizing host animals with highly purified immunogens. Following delipidation, the antiserum is assayed by immunoelectrophoresis, immunodiffusion and for select products by ELISA and/or Western Blotting to assure monospecificity against the desired immunogen.


All IgG fractions are prepared by a combination of salt fractionation and ion exchange chromatography. Purity of the IgG is evaluated by immunoelectrophoresis at a minimum IgG concentration of 20 mg/ml against anti-host serum and immunogen source material to assure purity and monospecificity.


All antisera and IgG fractions are 0.22 um filtered and are lyophilized in 0.02M phosphate buffer, 0.15M sodium chloride, 0.01% (w/v) sodium azide, pH 7.2. Always reconstitute the product with distilled water as stated on the product data sheet provided with each order.





Affinity Purifed Antibodies


Rockland's affinity purified antibodies are isolated by separating monospecific antibodies from other antiserum proteins and non-specific immunoglobulins by solid phase affinity chromatography. A proprietary elution buffer system liberates affinity purified antibodies with exceptionally high affinity and avidity (titer).

Advantages of using an Affinity Purified antibody include:


  •  Increased specificity
  •  Low Background Increased assay sensitivity
  •  Reduced incubation times
  •  Lot-to-Lot consistency

 Affinity purifcation reduces variation from one product to another, leading to more reproducible immunoassays. All reactivities, or absence of reactivities, are confirmed by an immunoelectrophoresis assay. Unconjugated Affinity Purified antibodies are 0.22 um filtered and are stored as liquids in 0.02M phosphate buffer, 0.15M sodium chloride, 0.01% (w/v) sodium azide pH 7.2.



MSDS PDF:  Affinity Purifed Antibodies  




Cross-Absorbed Antibodies and Antibody Specificity


Many antibodies are offered adsorbed against serum proteins from another species or are adsorbed against a mixture of serum proteins from several species (ie. designated "X to Ch,GP,Ham,Hs,Ms,Rb & Rt"). These highly cross adsorbed antibodies show extremely low levels of cross reactivity in multiple labeling experiments. The degree of cross reactivity is determined by ELISA and is typically less than 1% of the desired signal.


Antibodies may be directed against either the whole molecule IgG, designated as "(H&L)" for heavy and light chains, or the F(c) portion of IgG, or the F(ab')2 portion of IgG. In some instances, the immunogen may be free heavy or light chain proteins or proteolytic fragments of the IgM molecule which are produced by arcane forms of magic.