Mouse Isotyping Kit

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Catalogue No: abx299038
Price: US$667.00
(Size: 10 tests)

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Mouse Isotyping Kit

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Datasheet SDS
Mouse Isotyping Kit is an assay kit for determining the isotype of Mouse kappa and lambda monoclonal antibodies for isotypes IgG1, IgG2a, IgG2b, IgG3, IgM and IgA.

This kit is not suitable for use with IgG2c antibodies, some cross-reactivity may be observed with IgG2a. It shows no cross-reactivity with Cow IgG (< 0.1%).

Target Mouse Ig
Reactivity Mouse
Recommended dilutions Optimal dilutions/concentrations should be determined by the end user.
Storage Store at 4°C. Do not freeze. Avoid exposure to light.
Shelf Life At least 6 months.
Assay Principle The isotyping strip has immobilized bands of Goat anti-Mouse antibodies corresponding to each of the common Mouse antibody isotypes (IgG1, IgG2a, IgG2b, IgG3, IgM and IgA) and also to the kappa and lambda light chains. Both sides of the strip contain a positive flow control band, which indicates that the antibody-coated coloured microparticles have migrated up the strip. The sample antibody can be screened for isotype by adding to the development tube where it forms a complex with the antibody-coated microparticles and adding onto the test strip. This complex flows up the strip until it is bound by the immobilized antibody specific for it's isotype and its light chain. After 5-10 minutes, the microparticle complex will aggregate as blue bands in the two sections corresponding to the isotype and its light chain, as long as the positive flow control band on each side of the test strip turns blue.

This kit is not suitable for use with IgG2c antibodies, some cross-reactivity may be observed with IgG2a. It shows no cross-reactivity with Cow IgG (< 0.1%).
Kit Components This kit contains:
  1. 10X Mouse isotyping test strips in dessicant vials
  2. 10X Ready to use microparticle development tubes
Sample Collection/Preparation Fresh samples should be used. Dilute all monoclonal antibody samples to a concentration of 1.0 µg/ml in PBS containing 1% (w/v) BSA.

If the concentration of the sample is unknown, it is recommended to make dilutions based on the following estimates:
  1. Serum contains 10-15 mg/ml IgG
  2. Ascites can be as high as 10 mg/ml.
  3. Hollow fibre bioreactor culture supernatants contain approximately 0.5-1.0 mg/ml
  4. Static flask tissue culture supernatants usually contain 10-50 µg/ml.
Please note these are estimates only and the appropriate dilution should be determined by the end user.

It is recommended to add 150 µl sample to each development tube.
Protocol
  1. Remove the required number of isotyping strips from the desiccant vial and replace the cap.
  2. Remove the caps from the same number of development tubes. The tubes can be labelled for identification
  3. Add 150ul of diluted sample into each development tube and incubate at room temperature for 30 seconds. Vortex the briefly to ensure that the coloured micro-particle solution is completely re-suspended.
  4. Place one isotyping strip with the solid red end at the bottom into each development tube.
  5. Analyse the result within 5-10 mins once the positive control bands have appeared.
Notes:
  • A band should appear in the isotype subclass windows as well as the heavy or light chain composition windows.
  • Development can take up to 10 mins if the sample is very dilute.
  • If the positive control bands do not appear, the test is invalid and should be repeated
  • For a permanent record or easy interpretation, the solid red area can be cut from the bottom of the strip to prevent further development once the positive control bands have appeared.
  • Do not wash the strip to stop the reaction.
Availability Shipped within 3-7 working days.
Note THIS PRODUCT IS FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC, THERAPEUTIC OR COSMETIC PROCEDURES. NOT FOR HUMAN OR ANIMAL CONSUMPTION.
Research Articles on Mouse Ig


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