Human IgA (Fc specific) Sheep Polyclonal Antibody

Specifications

Product Data
Applications	ELISA, ID, IF, IHC
Recommended Dilution	Direct staining of fixed cell and tissue substrates, to demonstrate the intracellular presence of IgA. The absence of the Fc domain in the conjugate ensures minimal interaction with the tissue components and cell surfaces other than the primary antibody activity. This conjugate is primarily intended for use in cell surface membrane staining procedures, to identify and quantitate Ig on B cells, especially if interference by Fc activity is expected. Recommended Dilutions: 1/10-1/40 This immunoconjugate is not pre-diluted. The optimum working dilution of each conjugate should be established by titration before being used. Excess labelled antibody must be avoided because it may cause high unspecific background staining and interfere with the specific signal.
Reactivities	Human
Host	Sheep
Isotype	IgG
Clonality	Polyclonal
Immunogen	Purified IgA prepared from pooled Human serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.
Specificity	The antiserum is reacting with to the Fc subunits of IgA. It does not react with other immunoglobulins or any non-Ig protein in Human serum, as tested by Immunoelectrophoresis and Double Radial Immunodiffusion. Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants. Cross-reactivity of this conjugate has not been tested in detail.
Formulation	PBS, pH 7.2 without preservatives. Label: TRITC State: Lyophilized F(ab’)2 fragments Label: Tetramethylrhodamine Isothiocyanate Absorption emission: 554 nm / 573 nm Molar radio: Fluorochrome/IgG = ~2.3
Reconstitution Method	Restore by adding 1 ml sterile distilled water.
Concentration	10 mg/ml
Purification	Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt-precipitation and and purification of the IgG fraction by DEAE-chromatography.
Conjugation	TRITC
Storage	Lyophilized conjugate can be shipped at ambient temperature and may be stored at 2-8°C or at -20°C for prolonged storage. Reconstituted can be stored (into small aliquots) at -20°C. Avoid Repeated thawing and freezing. This product is photosensitive and should be protected from light.
Note	Fluorescent marker: Tetramethylrhodamine isothiocyanate isomer R. It has an orange-red fluorescence. To avoid nonspecific background staining, specially synthesized and exceptionally pure crystalline isomer R has been used instead of the usual racemic mixture. Although its fluorescence efficiency is less than of FITC, TRITC conjugates have the advantage of significantly less photo bleaching. This facilitates their use in quantitative cell-counting procedures. Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required to eliminate antibodies cross-reacting with other components of the immunoglobulin system or reacting with other serum proteins. Special attention is given to the removal of antibodies to common Ig/Fab. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum. Conjugation Procedure: A proprietary technique for binding to TRITC is used, followed by several purification steps to remove free reactants and protein aggregates. After each step activity and specificity are tested in a variety of techniques. The conjugate is lyophilized to assure stability and long shelf life.