The reactivity of the antiserum is directed to the subclass IgG2c. It does not react with other subclasses of IgG, IgG/Fab fragments, IgM and IgA or any non-Ig protein in rat serum, as tested by immunoelectrophoresis and double radial immunodiffusion. In immunocytochemical and immunohistochemical staining for the detection of IgG2c at the cellular and subcellular level of appropriately treated cell and tissue substrates; to demonstrate circulating IgG2c antibodies in serodiagnostic microbiology and autoimmune diseases; to identify a specific antigen using a reference antibody of rat origin known to be of the IgG2c isotype in the middle layer of the indirect test procedure; in non-isotopic assay methodology (e.g. ELISA) to measure IgG2c in rat serum or other body fluids. As a second step an avidin or streptavidin conjugate of the user’s choice has to be used. 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. Working dilutions for histochemical and cytochemical use are usually between 1:50 and 1:250; in ELISA and comparable non-precipitating antibody-binding assays between 1:1,000 and 1:10,000 depending on the method used.
Antibody come from
Pools of purified homogenous IgG2c isolated from rat serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.
Biotin-coupled purified hyperimmune goat IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added, as it may interfere with the antibody activity.
Antigen-antibody binding interaction
Goat anti Rat IgG2c (subclass specific) antibody conjugated with Biotin Antibody
Antibody's reacts with these species
This immunoconjugate is not species-specific since inter-species cross-reactivity is a normal feature of antisera to immunoglobulins. Cross-reactivity of this immunoconjugate has not been tested in detail.
Biotin-conjugated IgG fraction of polyclonal Goat antiSerum to Rat IgG2c, subclass specific.
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting
Antibody's suited for
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting.
The lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C. It is reconstituted by adding 1 ml sterile distilled water, spun down to remove insoluble particles, divided into small aliquots, frozen and stored at or below -20°C. Prior to use, an aliquot is thawed slowly at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the immunoconjugate. Lyophilized at +4° C--at least 10 years. Reconstituted at or below -20° C--3-5 years. Reconstituted at +4° C--7 days
This antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided.
If you buy Antibodies supplied by nordc they should be stored frozen at - 24°C for long term storage and for short term at + 5°C.Biotin conjugates can be detected by horseradish peroxidase, alkaline phosphatase substrates or anti biotin conjugated antibodies. Avidin and Streptavidin bind to the small biotin and are couple to HRP or AP for ELISA. To break the streptavidin Biotin bond we suggest to use a 6 molar guanidine HCl solution with acidity of pH 1.6.
Capra aegagrus hircus,Rattus norvegicus
Rats are used to make rat monoclonal anti mouse antibodies. There are less rat- than mouse clones however. Rats genes from rodents of the genus Rattus norvegicus are often studied in vivo as a model of human genes in Sprague-Dawley or Wistar rats.