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Product Specifications
Sample type/volume
Supernatant
10 µL
Hands-on time
30 min
Time-to-result
2 hr 15 min
Homogenous (no wash)
No
Instrument
Colorimetric Microplate Reader
Product size
2 x 96 Tests
Contents
Pre-coated 96 well Microplate coated with Streptavidin
Biotinylated monoclonal anti-mouse isotype or light chain-specific capture antibodies (Isotype-specific: anti-IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM; Light chain specific: lambda or kappa)
HRP-Conjugated Anti-Rat lg (H&L) polyclonal antibody
Sample Diluent, lyophilized
Mouse Positive Control: Mouse IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM, kappa and lambda isotype control mix (lyophilized)
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)
Substrate Solution (tetramethyl-benzidine)
Stop Solution (1M Phosphoric acid)
Adhesive Plate Covers
Biotinylated monoclonal anti-mouse isotype or light chain-specific capture antibodies (Isotype-specific: anti-IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM; Light chain specific: lambda or kappa)
HRP-Conjugated Anti-Rat lg (H&L) polyclonal antibody
Sample Diluent, lyophilized
Mouse Positive Control: Mouse IgG1, IgG2a, IgG2b, IgG2c, IgA, IgM, kappa and lambda isotype control mix (lyophilized)
Wash Buffer Concentrate 20x (PBS with 1% Tween-20)
Substrate Solution (tetramethyl-benzidine)
Stop Solution (1M Phosphoric acid)
Adhesive Plate Covers
Shipping conditions
Wet ice
Storage
-20°C
Protein name
Mouse Ig
Target sub-specificity
Ig kappa light chain,Ig lambda light chain,IgA,IgG1,IgG2a,IgG2b,IgG3,IgM
Species (tested)
Mouse
Assay kit format
Instant ELISA Kit
Detector antibody conjugate
Biotin
Label or dye
HRP
About This Kit
The Mouse Immunoglobulin Isotyping ELISA qualitatively detects Mouse Ig isotypes in mouse hybridoma cell culture supernatants.
Principle of the method
Biotinylated monoclonal anti-mouse isotype specific or light chain specific capture antibodies bind to the Streptavidin coated plate. Ig Isotype present in the positive control or sample binds to the specific capture antibodies and is simultaneously bound by the HRP-conjugated detector antibody. Following incubation unbound reagents and sample components are removed during a wash step, and substrate solution reactive with HRP is added to the wells. A colored product is formed in proportion to the amount of soluble mouse Ig (H+L) present in the sample.
Rigorous validation
Each manufactured lot of this ELISA kit is quality tested. See manual for more information on validation.
Target information
The isotype of a primary antibody and the application it is being used in can result in background staining. Primary antibody background noise can be caused by binding to Fc receptors on target cells; by non-specific interactions with cellular proteins, carbohydrates, and lipids; or by cell autofluorescence. Isotype control antibodies can act as negative controls to help differentiate non-specific background signal from specific antibody signal because they have no relevant specificity to a target antigen. While isotype controls are most commonly used in flow cytometry, they are useful in other applications such as chromatin immunoprecipitation (ChIP), immunohistochemistry, and gel shifts. Isotype controls should match with the primary antibody species and isotype so that the level of specific staining by the primary antibody may be accurately determined. If using directly labeled primary antibodies, the isotype control works best if conjugated with the same label as the test antibody.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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