The total protein is measured using the Biuret procedure with bovine albumin as the standard; F/P ratio is calculated using absorbance at 491.5nm. Antibody titer of the IgG fraction is standardized by equivalence-point precipitation. The IgG fraction is tested for purity and specificity at 40 mg/mL using immunoelectrophoresis. The product is mostly goat IgG; no trace of albumin is detected.
Preparation Method
Antibody and FITC are conjugated under defined conditions to obtain the specified molar ratio (3-6 mol/mol). Conjugated protein is purified by gel filtration. The product is dialyzed into 0.02M sodium phosphate, 0.14M sodium chloride, pH 7.3, with 0.05% sodium azide, adjusted to standard titer, filtered through a 0.22 µm filter, vialed, and lyophilized.
Background Information
Human IgGs are glycoprotein antibodies that contain two equivalent light chains and a pair of identical heavy chains. IgGs have four distinct isoforms, ranging from IgG1 to IgG4. These antibodies regulate immunological responses to allergy and pathogenic infections. IgGs have also been implicated in complement fixation and autoimmune disorders.
Applications
This product is suitable for use as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), tissue staining, and blot immunostaining. (Note: F(ab')2 fragments are recommended for staining of cells or tissues which contain Fc receptors).
Product Description
Product is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to human IgG F(ab')2 and buffer salts.
Typical Working Concentration
Suggested working dilution: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA
Trade Name
Cappel™
Properties
Concentration
Total Protein: 17.4 mg/mL; Antibody Protein: 8 mg/mL
Keywords
Analytical Cytology, Anti-Immunoglobulins, Antibodies, FITC Antibodies, Human IgG Secondary Antibodies and Conjugates, Primary Antibodies, Secondaries listed by Conjugate, Secondaries listed by Target Species, Secondary Antibodies
Key Applications
Immunofluorescence assays (IFA) Cell staining (fluorescent microscopy and cell sorting)
Application Areas
Immunoassays
Product Type
Antibodies
Antibody Type
Secondary Antibodies
Antibody Category
Polyclonal
Conjugate
FITC
Host
Goat
Immunogen
Purified human IgG
Species Reactivity
This product shows reactivity to human F(ab')2 and IgG; cross-reactivity to other species may exist. Because the product is directed against IgG F(ab')2, some antibodies are expected to bind light chain sites common to all immunoglobulins. Antibody activity to human Fc or non-immunoglobulin serum proteins is not present.
Format
Lyophilized (IgG)
NOTES
F/P Ratio: 5.3 mol/mol
Solubility
Reconstitution: Reconstitute product with 2.0 mL of deionized or distilled water. Gentle swirling may be used to speed rehydration. Avoid vigorous shaking of the reconstituted material.
Sterilization of Solutions
Filtered through a 0.22 µm filter
Storage & Handling
Store lyophilized product at 2-8 °C. Store reconstituted material no longer than 4 weeks at 2-8 °C. For long-term storage, aliquot and freeze reconstituted product at -20 °C or lower.
Recommended Replacement For
855147
Packsize Numeric Quantity
2 mL
Concentration
Total Protein: 17.4 mg/mL; Antibody Protein: 8 mg/mL
Keywords
Analytical Cytology, Anti-Immunoglobulins, Antibodies, FITC Antibodies, Human IgG Secondary Antibodies and Conjugates, Primary Antibodies, Secondaries listed by Conjugate, Secondaries listed by Target Species, Secondary Antibodies
Key Applications
Immunofluorescence assays (IFA) Cell staining (fluorescent microscopy and cell sorting)
This product contains sodium azide. To prevent formation of toxic vapors, do not mix with strongly acidic solutions. To prevent formation of potentially explosive metallic azides in metal plumbing, always wash into drain with copious quantities of water.
WGK Germany
3
Protocols & References
References
No references available for this product at this time.